Artesunate Combined with Other Medications and Action Mechanism： A Review / 中国实验方剂学杂志

A sesquiterpene natural substance called artemisinin was discovered in Artemisia annua. One of its derivatives， artesunate （ART）， has the properties of economy， immediate effect， low toxicity， and good tolerance. Since it has a quick and powerful killing effect on plasmodium in the erythrocyte phase and can quickly handle clinical seizure and symptoms， it is currently mostly utilized to treat cerebral malaria and other severe instances of malaria. In addition， it has antitumor， antivirus， anti-hepatic fibrosis， anti-inflammatory， antibacterial， hepatocyte protection， immunological modulation， and other pharmacological properties and can inhibit cell proliferation， induce cell apoptosis， and reduce the incidence of sepsis. In many countries， artemisinin-based combination therapies （ACTs）， such as artemether-benflumetol， artesunate-amodiaquine， and artemether-lumefantrine， are the first-line treatments for malaria. Recent research on artesunate by Chinese and international scholars has revealed that compared with monotherapy， artesunate combination therapy offers more benefits in terms of improving pharmacological effects， shortening the duration of medicine， and minimizing adverse effects. Through systematic retrieval of Web of Science Core Collection and integration through CiteSpace （6.2.1） software， this article reviewed the mechanism of artesunate combined with other medications with regard to antimalarial， antitumor， antibacterial， and antiviral features in the previous five years， so as to provide some theoretical basis for rational development and utilization of ART and new drug research and development.

Mechanism of artesunate on bone destruction in experimental rheumatoid arthritis based on transcriptomics and network pharmacology / 中国中药杂志

The present study investigated the mechanism of artesunate in the treatment of bone destruction in experimental rheumatoid arthritis(RA) based on transcriptomics and network pharmacology. The transcriptome sequencing data of artesunate in the inhibition of osteoclast differentiation were analyzed to obtain differentially expressed genes(DEGs). GraphPad Prism 8 software was used to plot volcano maps and heat maps were plotted through the website of bioinformatics. GeneCards and OMIM were used to collect information on key targets of bone destruction in RA. The DEGs of artesunate in inhibiting osteoclast differentiation and key target genes of bone destruction in RA were intersected by the Venny 2.1.0 platform, and the intersection target genes were analyzed by Gene Ontology(GO)/Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment. Finally, the receptor activator of nuclear factor-κB(RANKL)-induced osteoclast differentiation model and collagen-induced arthritis(CIA) model were established. Quantitative real time polymerase chain reaction(q-PCR), immunofluorescence, and immunohistochemistry were used to verify the pharmacological effect and molecular mechanism of artesunate in the treatment of bone destruction in RA. In this study, the RANKL-induced osteoclast differentiation model in vitro was established and intervened with artesunate, and transcriptome sequencing data were analyzed to obtain 744 DEGs of artesunate in inhibiting osteoclast differentiation. A total of 1 291 major target genes of bone destruction in RA were obtained from GeneCards and OMIM. The target genes of artesunate in inhibiting osteoclast differentiation and the target genes of bone destruction in RA were intersected to obtain 61 target genes of artesunate against bone destruction in RA. The intersected target genes were analyzed by GO/KEGG enrichment. According to the results previously reported, the cytokine-cytokine receptor interaction signaling pathway was selected for experimental verification. Artesunate intervention in the RANKL-induced osteoclast differentiation model showed that artesunate inhibited CC chemokine receptor 3(CCR3), CC chemokine receptor 1(CCR1) and leukemia inhibitory factor(LIF) mRNA expression in osteoclasts in a dose-dependent manner compared with the RANKL-induced group. Meanwhile, the results of immunofluorescence and immunohistochemistry showed that artesunate could dose-dependently reduce the expression of CCR3 in osteoclasts and joint tissues of the CIA rat model in vitro. This study indicated that artesunate regulated the CCR3 in the cytokine-cytokine receptor interaction signaling pathway in the treatment of bone destruction in RA and provided a new target gene for the treatment of bone destruction in RA.

Exploring the potential mechanism of artesunate in intervening with U87 cells and U251 cells with distinct therapeutic effects on the basis of transcriptome sequencing and network pharmacology / 药学学报

Artesunate possesses the potential of intervening with glioma, however, its pharmacological mechanisms remain unclarified. Firstly, the effects of artesunate on cell activity, proliferation and apoptosis of U87 and U251 human glioma cells were explored. It was found that artesunate exerted stronger inhibitory effects on the activity and proliferation of U87 cells than U251 cells. It could significantly promote apoptosis in U87 cells (P < 0.05), while only high dose of artesunate can promote that of U251 cells (P < 0.01), detected by Hoechst and TUNEL cell apoptosis staining. Further, the differential expression gene sets between artesunate-sensitive and non-sensitive cell line, as well the therapeutic effects-related genes of artesunate were obtained through transcriptome sequencing and differential data analysis by using the lysates of U87 and U251 cells before and after artesunate treatment, aiming to explore the molecular mechanism of distinct artesunate sensitivity to two types of cells. Then, key putative targets that related to therapeutic effects were screened by constructing the interaction network of differential genes of three above comparison groups, and calculating their topological characteristics. Pathway enrichment analysis showed that those key putative targets were significantly enriched in several signaling pathways that were closely associated with the main pathological changes of glioma, among which apoptosis-related activating transcription factor 4 (ATF4)-DNA damage induced transcript 3 (DDIT3)- polyadenosine diphosphate ribose polymerase 1 (PARP1) signaling axis was the most enriched in. Molecular docking indicated that artesunate had fine binding affinities with ATF4 and DDIT3. Above all, this study preliminarily revealed that ATF4-DDIT3-PARP1 signaling axis is the target pathway of artesunate intervening with U87 glioma cells, and PARP1 may be an important gene for U251 cells to develop resistance to artesunate. Our results not only provide fundamental experimental evidence for artesunate as a potential therapeutic drug in glioma treatment, but shed light into overcoming drug resistance in its clinical therapy.

Anti-inflammatory Effect and Mechanism of Artesunate： A Review / 中国实验方剂学杂志

Inflammation， the basic pathological process of many diseases， can occur in various tissues and organs of the body and cause many diseases including cancer. So far， there are thousands of anti-inflammatory drugs on the market， but most of these drugs have adverse reactions of gastrointestinal injury， and can even cause greater damage to the body. In recent years， the research on the repurpose of Chinese medicine is in the ascendant， and the innovative research on the specific antimalarial drug artemisinin has attracted extensive attention from scholars in China and abroad. Artesunate is a water-soluble derivative of artemisinin， which has the characteristics of quick effect and low toxicity. In addition to its significant therapeutic effect on malaria， artesunate also has a potential anti-inflammatory effect. In this review， the anti-inflammatory effect and mechanism of artesunate were elaborated in detail by consulting the relevant literature. It was found that artesunate had good anti-inflammatory effects in the respiratory system， liver injury， osteoarthritis， dermatitis， kidney inflammation， colitis， neuroinflammation， and even in novel coronavirus disease 2019 （COVID-19）. It was concluded that artesunate mainly participated in apoptotic signal transduction， mediated immune regulation， and improved oxidative stress to play an anti-inflammatory role by acting on nuclear factor-κB （NF-κB）， nuclear factor E2-related factor 2 （Nrf2）， phosphatidylinositol-3 kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR）， Toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/tumor necrosis factor receptor-associated factor 6 （TRAF6）， high mobility group box 1 （HMGB1）/receptor for advanced glycation endproduct （RAGE）， and other pathways. Through the review of the anti-inflammatory effect and mechanism of artesunate， it is expected to provide a reference for the application of artesunate in inflammation resistance and further development and utilization of artesunate in the future.

Establishment of Poly （I∶C）-induced Cytokine Storm Model in Mice and Intervention Effect of Artesunate and Qingfei Paidu Decoction / 中国实验方剂学杂志

ObjectiveTo observe the intervention effect of artesunate （ART） and Qingfei Paidu decoction （QFPD） on the mouse model of cytokine storm （CS） induced by viral mimic Poly （I∶C）. MethodEighty-four SPF male BALB/c mice were randomly divided into seven groups， with 12 mice in each group. Mice， except for those in the blank group （n=12）， were subjected to CS model induction by tail vein injection of Poly （I∶C） at 15 mg·kg-1， followed by drug treatments of low-dose ART （ART-l， 10 mg·kg-1）， medium-dose ART （ART-m， 20 mg·kg-1）， high-dose ART （ART-h， 40 mg·kg-1）， Qingfei Paidu Decoction （QFPD， 2.4 g·kg-1）， and dexamethasone （DXM， 10 mg·kg-1）. After 6 hours， lung tissues， bronchoalveolar lavage fluid （BALF）， spleen， lung， and peripheral blood were collected. The lung and spleen indexes were calculated and the number of inflammatory cells in BALF was detected. The pathological changes in lung tissues were observed by hematoxylin-eosin （HE） staining and the levels of tumor necrosis factor-α （TNF-α）， interferon-γ （IFN-γ）， interleukin-1β （IL-1β）， and IL-6 in BALF were detected by enzyme-linked immunosorbent assay （ELISA）. The expression of immune cells in BALF and peripheral blood was detected by flow cytometry. ResultThe analysis of lung and spleen indexes showed that compared with the blank group， the model group showed increased lung and spleen indexes to varying degrees （P<0.05）. Compared with the model group， the ART groups showed reduced spleen index （P<0.05） and the ART-l group showed reduced lung index （P<0.05）. Additionally， the QFPD group showed reduced lung and spleen indexes （P<0.05）. ELISA results showed that except for TNF-α， the levels of IFN-γ， IL-1β， and IL-6 in the model group increased compared with those in the blank group （P<0.05）. Compared with the model group， the ART-l group and the QFPD group showed reduced content of TNF-α （P<0.05）， and all groups with drug intervention showed reduced content of IFN-γ， IL-1β， and IL-6 （P<0.05）. The number of inflammatory cells in BALF showed a downward trend in the model group， and the number of cells increased in the groups with drug intervention except for the DXM group （P<0.05）. Flow cytometry showed that compared with the blank group， the model group showed decreased number of CD3 in the peripheral blood （P<0.05）， increased Ly-6G and F4/80 （P<0.05）， decreased expression of CD45， CD3， and F4/80 in BALF （P<0.05）， and increased expressions of Ly-6G （P<0.05）. Compared with the model group， the ART groups and QFPD group showed increased CD45 content in peripheral blood （P<0.05）， decreased Ly-6G and F4/80 content （P<0.05）， increased CD45 and F4/80 content in BALF （P<0.05）， and decreased expression of Ly-6G （P<0.05）. ConclusionART and QFPD have a good protective effect on Poly （I∶C）-induced CS in mice， and the mechanism is related to the effective intervention in immune cell disorder.

Artesunate reduces neuronal apoptosis and inflammatory response in model rats with ischemic stroke in vivo, and promotes microglia polarization in vitro / 中华行为医学与脑科学杂志

Objective:To investigate the effects of artesunate ( ART ) on neuronal apoptosis, inflammatory response after stroke in rats and microglia polarization.Methods:(1)Animal experiment: twenty-seven male SD rats of SPF grade were divided into sham operation group, model group and ART treatment group according to the random number table method, with 9 rats in each group.Rats in the model group and ART treatment group were used to establish a stroke model by middle cerebral artery occlusion (MCAO). And rats in the ART treatment group were intraperitoneally injected with ART (25 mg/kg) once a day for three days before modeling, while the rats in sham operation group and the model group were injected with the same amount of solvent.And 24 h after the modeling, TTC staining was used to evaluate the volume of cerebral infarction, Western blot was used to detect the expression of Bcl2 in the infarct area, penumbra and hippocampus, TUNEL method was used to detect neuronal apoptosis, and tissue immunofluorescence was used to observe the expression of tumor necrosis factor-α(TNF-α) in the penumbra region of cerebral cortex.(2)Cell experiments: microglia BV2 were cultured and divided into control group, oxygen-glucose deprivation/reoxygenation group, oxygen-glucose deprivation/reoxygenation + 0.05 μmol/L ART group, oxygen-glucose deprivation/reoxygenation + 0.1 μmol/L ART group and oxygen-glucose deprivation/reoxygenation + 0.5 μmol/L ART group.The levels of inflammatory factors interleukin-6(IL-6), interleukin-1β(IL-1β) and TNF-α were detected by qRT-PCR, the expressions of M2 type microglia marker protein CD206 and ARG1 were detected by Western blot, the BV2 cell medium after treatment in each of the above groups was collected as conditioned medium to culture HT22 hippocampal neuron cells and cell activity was measured by CCK8 method.GraphPad Prism 7 software was used for data analysis.One-way ANOVA was used for comparison of differences among multiple groups, and LSD was used for further two-by-two comparisons.Results:(1)Animal experiment results: TTC staining results showed that the percentage of cerebral infarction volume in the ART treatment group was smaller than that in the model group ((23.09±8.51)%, (39.63±5.71)%, t=33.93, P<0.01). The results of TUNEL staining showed that the number of apoptotic cells in the model group and ART treatment group was higher than that in the sham operation group ((638.90±177.82)cells/mm 2, (72.75±13.21) cells/mm 2, (16.16±2.73) cells/mm 2, both P<0.05), and the number of apoptotic cells in the ART treatment group was lower than that in the model group ( P<0.05). Western blot results showed that the levels of Bcl2 protein in penumbra and infarct area of the model group were both lower than those in sham group(both P<0.05). The levels of Bcl2 protein in penumbra, the hippocampus and infarcted area of the ART treatment group were significantly lower than those of the model group(all P<0.05). The results of tissue immunofluorescence showed that the fluorescence intensities of TNF-α in the model group and ART treatment group were higher than those in the sham group (all P<0.05), while the fluorescence intensity of TNF-α in the ART treatment group was lower than that in the model group ( P<0.05). (2)Cell experiment: qRT-PCR results showed that compared with the control group, the mRNA levels of IL-6, IL-1β and TNF-α (all P<0.05) in oxygen-glucose deprivation/reoxygenation group were significantly higher than those of the control group.And the mRNA levels of IL-1β, IL-6 and TNF-α in oxygen-glucose deprivation/reoxygenation + 0.05 μmol/L ART group, oxygen-glucose deprivation/reoxygenation + 0.1 μmol/L ART group and oxygen-glucose deprivation/reoxygenation + 0.5 μmol/L ART group were significantly lower than those of the oxygen-glucose deprivation/reoxygenation group (all P<0.05). Western blot results showed that compared with the control group, the expression of CD206 ((0.85±0.04), (1.07±0.07), P<0.05) was significantly down-regulated in the oxygen-glucose deprivation/reoxygenation group.The CD206 and ARG in oxygen-glucose deprivation/reoxygenation + 0.1 μmol/L ART group((1.22±0.06), (1.35±0.08)) and oxygen-glucose deprivation/reoxygenation + 0.5 μmol/L ART group((1.24±0.14), (1.14±0.07)) were significantly higer than those of oxygen-glucose deprivation/reoxygenation group((0.85±0.04), (0.85±0.05))(all P<0.05). The results of CCK8 showed that compared with the control group, the cell viability in the oxygen-glucose deprivation/reoxygenation group was significantly decreased( P<0.05). The cell viability of the oxygen-glucose deprivation/reoxygenation + 0.05 μmol/L ART group, the oxygen-glucose deprivation/reoxygenation + 0.1 μmol/L ART group, the oxygen-glucose deprivation/reoxygenation + 0.5 μmol/L ART group were all higher than those of oxygen-glucose deprivation/reoxygenation group(all P<0.05). Conclusion:ART reduces neuronal apoptosis after stroke, decreases the neuroinflammatory response after stroke, and promotes oxygen-glucose deprivation/reoxygenation-activated microglia BV2 polarization to the M2 type.

Comparative, observational study of the use of artesunate injections along with standard-of-care treatment versus only standard-of-care treatment in moderate and severe acute respiratory distress syndrome cases of COVID-19-positive infections

Introduction: COVID-19 is a type of coronavirus disease belonging to the family Coronaviridae. In late December 2019, this virus emerged from Wuhan, Hubei province, China, and resulted in an outbreak in China and expanded globally. In India, the mortality rate today was 521,691 till the date-time of writing this article. Several therapeutic agents have been evaluated for the treatment of COVID-19. Materials and Methods: This was a hospital-based comparative, observational study of the use of artesunate injections with standard-of-care (SOC) treatment (group A) versus only SOC (group B) treatment in moderate- to-severe cases of COVID-19 acute respiratory distress syndrome (ARDS) patients, on a total of 130 patients (comparative group of 65 patients each). The study was done on hospitalized COVID-19-positive moderate and severe cases of ARDS from October 2020 to June 2021 at MGM Hospital and Research Centre, CBD Belapur, Navi Mumbai, Maharashtra, India. Results: One hundred and thirty patients were divided into two groups of 65 each; group A was compared with group B; group A received SOC with artesunate injections and group B received only SOC treatment. The mean age of patients in group A was 57.3 ± 12.5 years (standard deviation [SD]: 54.2–60.3) and in group B was 55.8 ± 12.5 years (SD: 52.8–58.9). Diabetes mellitus was the most comorbid condition. The inflammatory markers, respiratory rate, and SpO2 improved in group A as compared to group B. The proportion of patients progressing to noninvasive and invasive ventilation was more in group B as compared to group A (P < 0.05). About 93.8% of patients (61 patients) recovered in group A compared with 72.3% of patients (47 patients) who recovered in group B. The overall death in group A was 6.2% (four patients) and 27.7% (18 patients) in group B (P < 0.05), indicating the proportion of dead patients is significantly more where only SOC treatment was given. Conclusions: Artesunate injection administration accelerated recovery in our patients with moderate and severe COVID-19 disease by controlling hyperimmune response. The clinical improvement was seen by decreased levels of inflammatory markers, reduced respiratory rate, and improved oxygen saturation and showed significant survival in group A compared with group B. Artesunate injections were given 2 mg/kg body weight diluted in 1 mL 5% sodium bicarbonate solution as a bolus followed by 1 mg/kg body weight after 6 h and 2 mg/kg body weight with 1 mL sodium bicarbonate solution for next 2 days at an interval of 24 h. Patients tolerated the injections well and recovery improved, so artesunate can be considered a therapeutic option in moderate and severe cases of COVID-19 ARDS.

Manejo clínico-terapéutico de la malaria grave

La malaria representa un grave problema de salud pública en el país, por su morbilidad y mortalidad. Es importante conocer la patogenia y las manifestaciones clínicas de la malaria grave, en especial revisar el ciclo biológico del parásito, ya que la enfermedad comienza con la ruptura del esquizonte maduro, siendo las primeras manifestaciones clínicas: fiebre y anemia. La infección por Plasmodium falciparum es más severa y es mediada por el fenómeno de secuestro en la microvasculatura venosa profunda, mientras que Plasmodium vivax causa una enfermedad debilitante, rara vez mortal, pero en oportunidades se presentan manifestaciones graves que causan la muerte del paciente. Malaria grave se define por la presencia de signos clínicos y de laboratorio de disfunción de órganos vitales como sistema nervioso central, riñón, gastrointestinal, vías respiratorias y alteraciones hemodinámicas; la cual requiere el rápido reconocimiento de la enfermedad y del grado de severidad. Se debe hacer un manejo de índole general y prestar especial atención a la terapia antimalárica oportuna con Artesunato, primera línea en malaria grave, o Arthemeter o Quinina con Clindamicina según los protocolos nacionales e internacionales, para lograr una evolución satisfactoria. En consecuencia, es un reto enfrentar esta entidad y obliga a la constante actualización en las diferentes opciones cónsonas con las diferentes especies de Plasmodium patógeno.

Malaria represents a serious public health problem in the country, due to its morbidity and mortality. It is of most importance to know the pathogenesis and clinical manifestations of severe malaria, particularly to review the biological cycle of the parasite. The disease begins with the rupture of the mature schizont, with the first clinical manifestations being fever and anemia. Plasmodium falciparum infection is more severe and is mediated by the phenomenon of sequestration in the deep venous microvasculature, while Plasmodium vivax causes a debilitating disease, rarely fatal, but sometimes serious manifestations occur that cause the death of the patient. Severe malaria is defined by the presence of clinical and laboratory signs of dysfunction of vital organs such as the central nervous system, kidney, gastrointestinal, respiratory tract, and pathological hemodynamic changes that requires rapid disease recognition and degree of severity. General management and timely antimalarial therapy with Artesunate, first line in severe malaria, or Arthemeter, or Quinine with Clindamycin following national and international protocols, achieve a favorable outcome. Consequently, it is a challenge to face this entity and requires constant updating in the different options consistent with the different species of pathogenic Plasmodium.

Exploring the effects of artesunate and fuzheng huayu decoction on mitochondria in the treatment of schistosomiasis liver fibrosis / 中华肝脏病杂志

Objective: To compare the effects of artesunate (Art) and fuzheng huayu decoction on mitochondrial autophagy in the treatment of schistosomiasis liver fibrosis. Methods: Eighty C57BL/6 female mice were randomly divided into healthy control group, infection group, Art treatment group and Fuzheng Huayu Decoction treatment group, with 20 mice in each group. Mice in the infection group and treatment group were infected with 16 Schistosoma japonicum cercariae. After 6 weeks, praziquantel (300 mg/kg) was used for 2 days to kill the worms. The Art treatment group was treated with intraperitoneal injection of 100 mg/kg/day, while the Fuzheng Huayu Decoction treatment group was fed 16g of fuzheng huayu decoction per 1kg per day. After 6 weeks, fresh liver tissues of the four groups were collected. Masson staining and Western blot were used to observe the succinate dehydrogenase subunit A (SDHA) and malate dehydrogenase (MDH2), citrate synthase (CS), ketoglutarate dehydrogenase (OGDH), and target of rapamycin 1 (mTORC1) pathway involved in mitochondrial tricarboxylic acid cycle in liver tissues. The relative expression levels of adenylate activated protein kinase (AMPK) and mitochondrial autophagy pathway kinase (PINK1) were detected. Liver tissue samples were extracted from each group to detect the mitochondrial oxygen consumption rate. Two-way ANOVA was used to compare the significance and difference between two sets of samples. Results: Masson staining showed that the infection group mice had significantly higher liver fibrosis area than the healthy control group, while the Art treatment group and Fuzheng Huayu Decoction treatment group mice had lower liver fibrosis area than the infection group. Western blot analysis showed that the infection group (0.82 ± 0.05) had significantly lower relative expression of SDHA protein than the healthy control group (1.00 ± 0.05) (t = 11.23, P = 0.0035), while the Art treatment group (0.73 ± 0.05) had significantly higher relative expression of SDHA protein than the infection group (t = 10.79, P = 0.0073). However, there was no significant change in Fuzheng Huayu Decoction treatment group (0.98±0.05) (t = 1.925, P = 0.1266). The relative expression of p-AMPK protein was significantly higher in the infection group (1.15 ±0.05) than in the healthy control group (0.98 ± 0.07, t = 12.18, P = 0.0029), and the expression of p-AMPK in the Art treatment group (0.50 ± 0.05) was significantly lower than the infection group (t = 11.78, P = 0.0032). The relative protein expression of AMPK was significantly lower in the infection group (0.80 ± 0.05) than in the healthy control group (1.00 ± 0.05, t = 10.53, P = 0.0046). The expression of AMPK was significantly lower in the Art treatment group (0.54 ± 0.05) than in the infection group (T = 13.98, P = 0.0036). The relative expression of p-mTORC1 protein (0.93 ± 0.08) was not significantly different in the infection group than in the healthy control group (t = 2.28, P = 0.065), while the Art treatment group (0.63 ± 0.05) had significantly lower relative expression of p-mTORC1 protein than the infection group (t = 10.58, P = 0.029). The expression of p-mTORC1/ m-TORC1 was not significantly different in the infection group (0.98 ± 0.03) than in the healthy control group (0.97 ± 0.03, t = 0.98, P = 0.085), while the Art treatment group (0.63 ± 0.05) had significantly lower relative expression of p-mTORC1/ m-TORC1 than the infection group (t = 14.58, P = 0. 009). The relative protein expression of PINK1 was significantly lower in the infection group (0.55 ± 0.05) than in the healthy control group (1.00 ± 0.03, t = 13.49, P = 0.0011), while the Art treatment group (1.21 ± 0.05, t = 9.98, P = 0.0046) and Fuzheng Huayu Decoction treatment group (1.31 ±0.35, t = 6.98, P = 0.027) had significantly higher relative protein expression of PINK1 than the infection group. Mitochondrial function tests showed that after adding substrate complex II, the oxygen consumption of the infection group was lower than the healthy control group, while the Art treatment group and the Fuzheng Huayu Decoction treatment group had higher oxygen consumption than the infection group. The oxygen consumption was significantly lower after adding the substrate complex III in the infection group than the healthy control group, while the Art treatment group and Fuzheng Huayu Decoction treatment group had higher oxygen consumption than the infection group. Conclusion: Art can alleviate schistosomiasis liver fibrosis by inhibiting AMPK/mTORC1 signaling pathway activity and enhancing mitochondrial oxygen consumption, autophagy and SDHA expression.

Therapeutic effect of artesunate on acute Toxoplasma gondii infection in mice / 中华地方病学杂志

Objective:To investigate the therapeutic effect of artesunate on mice with acute Toxoplasma gondii ( T. gondii) infection. Methods:Based on body weight (16 - 18 g), sixty-four C57BL/6 female mice aged 6 - 8 weeks were divided into 4 groups by random number table method: uninfected control group without treatment; T. gondii-infected group (Tg group), each mouse was intraperitoneally (i.p.) infected with 100 tachyzoites of T. gondii RH strain; T. gondii-infected + artesunate treatment group (Tg + ART group), 3 hours after each mouse i.p. infected with 100 tachyzoites of T. gondii RH strain, artesunate solution was i.p. injected at a dose of 30 mg/kg, once a day for a total of 7 consecutive days; T. gondii-infected + sulfadiazine treatment group (Tg + SDZ group), 3 hours after each mouse i.p. infected with 100 tachyzoites of T. gondii RH strain, sulfadiazine solution was orally administrated at a dose of 100 mg/kg, once a day for a total of 7 consecutive days. There were 16 mice in each group, in which 10 mice were used to observe survival time and 6 mice were used to monitor body weight and collect tissue samples. Mice were weighed every day from day 1 post infection (p.i.); mice were sacrificed at day 7 p.i., the liver weights of mice were weighed and the liver indexes were calculated; liver tissues were paraffin-embedded, sectioned, and stained with hematoxylin-eosin (HE), and the pathological changes of liver tissues of mice in each group were observed under a light microscope. The expression levels of T. gondii major surface antigen 1 (SAG1) in the liver tissues of mice in each group were detected by real-time quantitative PCR for evaluating parasite load. Results:All mice in the uninfected control group were survived. The survival time was 7 - 9 days in Tg group, 8 - 11 days in Tg + ART group, and 9 - 13 days in Tg + SDZ group. Compared with Tg group, the survival times of mice in Tg + ART group and Tg + SDZ group were significantly longer ( P < 0.05). On day 7 p.i., compared with uninfected control group, Tg + ART group or Tg + SDZ group, the body weight of mice in Tg group was lower ( P < 0.05); however, there was no significant difference of body weight in Tg + ART group and Tg + SDZ group compared with uninfected control group ( P > 0.05). Compared with Tg group, Tg + ART group and Tg + SDZ group had lower liver indexes and SAG1 mRNA expression levels in the liver tissues ( P < 0.05 or < 0.001), and liver histopathological changes were milder. Compared with Tg + SDZ group, there was no significant difference in both liver index and SAG1 mRNA expression level in the liver tissue of Tg + ART group ( P > 0.05). Conclusion:Artesunate solution i.p. injection can prolong the survival time, reduce parasite load in the liver, and attenuate hepatic pathological damage, to a certain extent, of mice with acute T. gondii infection.

Role of three drugs in traumatic proliferative vitreoretinopathy in rabbits / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the effect of triamcinolone acetonide(TA), artesunate(ART), and luteolin(LU)on the prevention and treatment of traumatic proliferative vitreoretinopathy(TPVR). <p>METHODS: Forty-eight cyanotic blue rabbits were selected to prepare TPVR animal models by making a penetrating eye injury and intravitreal injection of 0.3mL platelet-rich plasma, and were randomly divided into four groups(<i>n</i>=12), in which the vitreous cavity of the control group was injected with 0.1mL saline; The vitreous cavity of the TA group was injected with 0.1mL(1mg/mL)triamcinolone acetonide; The vitreous cavity of the ART group was injected with 0.1mL(20μg/mL)artesunate; 0.1mL(10μg/mL)luteolin was injected into the vitreous cavity of the LU group. The vitreous and retinal proliferation were observed by fundus photography and ocular ultrasound at 1, 2, 3 and 4wk postoperatively. The expression levels of α-SMA and VIM protein in the vitreous fluid of each group of rabbit eyes were detected by Western Blot at 28d postoperatively, and the retinal tissue structure of each group was observed by retinal HE staining. <p>RESULTS: At 28d postoperatively, the TPVR grading of rabbit eyes in the TA, ART and LU groups were significantly lower than that in the control group(<i>P</i><0.05), and the TPVR grading of rabbit eyes in the TA group was significantly lower than that in the ART and LU groups(<i>P</i><0.05). The expression levels of α-SMA and VIM proteins in the vitreous fluid of the rabbit eyes in the TA, ART and LU groups were significantly lower than those in the control group at 28d after surgery(<i>P</i><0.01). The results of HE staining showed that the arrangement of retinal layers in rabbit eyesin the control group were disordered, severely distorted or locally broken, the structure of each layer were unclear, the anterior membrane was obviously thickened, and the retina was obviously detached; The arrangement of retinal layersin rabbit eyes in the LU group were slightly distorted, inflammatory exudation was visible in front of the retina, and the retina was superficially detached; The structure of retina in rabbit eyes in the ART group were clear, with mild edema and superficial detachment; The structure of retinal layers in rabbit eyes in the TA group were clear, the arrangement was still neat, the retinal folds were locally visible, and there was no retinal detachment.<p>CONCLUSION: Intravitreal injection of triamcinolone acetonide, artesunate and luteolin were all effective in preventing and treating traumatic TPVR, among which triamcinolone acetonide has the most obvious effect.

Effect of artesunate on intestinal ischemia/reperfusion-induced lung injury in mice and relationship with heme oxygenase-1 / 中华麻醉学杂志

Objective:To evaluate the effect of artesunate on intestinal ischemia/reperfusion (I/R)-induced lung injury in mice and relationship with heme oxygenase-1 (HO-1).Methods:Twenty-four healthy SPF male C57BL/6 mice, aged 8-9 weeks, weighing 18-22 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (group Sham), intestinal I/R group (group I/R), artesunate group (group A), and artesunate plus HO-1 inhibitor Zinc protoporphyrin Ⅸ(ZnPP) group (group AS). The model of intestinal I/R injury was established by occluding the superior mesenteric artery for 45 min followed by 2 h reperfusion in anesthetized animals.Artesunate 40 mg/kg was injected via the tail vein at 1 h before ischemia in group A. ZnPP 7.5 mg/kg was injected via the tail vein at 12 h before ischemia, and artesunate 40 mg/kg was injected via the tail vein at 1 h before ischemia in group AS.The animals were sacrificed at the end of reperfusion, and the lung tissues were obtained for microscopic examination of the pathologic changes and for determination of the wet/dry lung weight (W/D) ratio, myeloperoxidase (MPO) activity, malondialdehyde (MDA) content, expression of interleukin-6 (IL-6) mRNA (by fluorescence quantitative polymerase chain reaction) and apoptotic index (AI) (by TUNEL). The lung injury score was assessed. Results:Compared with group Sham, the lung injury score, W/D ratio, MPO activity, MDA content and AI were significantly increased, and the expression of IL-6 mRNA was up-regulated in group I/R ( P<0.05). Compared with group I/R, the lung injury score, W/D ratio, MPO activity, MDA content and AI were significantly decreased, and the expression of IL-6 mRNA was down-regulated in group A ( P<0.05). Compared with group A, the lung injury score, W/D ratio, MPO activity, MDA content and AI were significantly increased, and the expression of IL-6 mRNA was up-regulated in group AS ( P<0.05). Conclusions:Artesunate can alleviate intestinal I/R-induced lung injury, and the mechanism may be related to activation of HO-1 in mice.

Mechanism of Artesunate in Reversing Epithelial-mesenchymal Transition and Akt /Snail Signaling Pathway in Colorectal Carcinoma / 中国实验方剂学杂志

Objective:To investigate the effects of artesunate （ART） on epithelial-mesenchymal transformation （EMT） of colorectal cancer HCT-8 cells，and explore the effects of ART on cell migration，invasion，EMT ability， and protein kinase B （Akt）/Snail signaling pathway of colorectal cancer. Method:3-（4-5-Dimethylthiazol-2-yl）2，5-diphenyltetrazolium bromide （MTT） assay was used to detect the effects of ART at different concentrations on the proliferation of HCT-8 cells. Wound healing assay and Transwell assay were used respectively to detect the effects of ART on migration and invasion of colorectal cancer cells. The effects of different concentrations of ART on the distribution of EMT-related proteins vimentin and E-cadherin in HCT-8 cells were detected by double-immunofluorescent staining. The effects of ART on protein expression levels of EMT markers E-cadherin，vimentin and N-cadherin in HCT-8 cells and the expression of Akt1， p-Akt1， and Snail1 in the Akt/Snail signaling pathway were determined by Western blot. Result:The dose-dependent inhibitory effects of ART on the proliferation of HCT-8 cells were determined and the inhibition rate was calculated. A dose-response curve was plotted accordingly. The half-maximal inhibitory concentration （IC₅₀） of ART on HCT-8 cells was （16.67±1.95） μmol·L^-1. The following four groups were set up： a control group （0 μmol·L^-1），and low-， medium-， and high-dose ART groups（2， 10， 50 μmol·L^-1）. Compared with the results in the control group，ART inhibited the migration and invasion of HCT-8 cells（<italic>P</italic><0.05）. Specifically， the expression of E-cadherin in HCT-8 cells was significantly up-regulated，and that of vimentin and N-cadherin was significantly down-regulated （<italic>P</italic><0.05）. The expression levels of p-Akt1 and Snail1 were significantly decreased after ART treatment，thus inhibiting EMT（<italic>P</italic><0.05）. Conclusion:The findings of this study suggested that ART inhibited the EMT-triggered migration and invasion of HCT-8 cells presumedly by inhibiting the activation of the Akt/Snail pathway to reverse EMT.

Effect of Artesunate on Experimental Choroidal Neovascularization and Expression of HIF-1α and VEGF / 中国实验方剂学杂志

Objective:To observe the effects of artesunate （ART） on experimental choroidal neovascularization （CNV） and the expression of related proteins， and to explore the underlying mechanism. Method:Eighty BN rats were randomly divided into five groups： a normal group， a model group， a conbercept group， and low- （10.08 mg·kg^-1·d^-1） and high-dose （20.16 mg·kg^-1·d^-1） ART groups， with 16 rats in each group. A CNV model was established with 532 nm laser in rats of the groups except for the normal group. The rats in each group were treated with corresponding drugs by gavage for 14 days. The normal group， the model group， and the conbercept group received 1% CMC-Na solution at the same volume， while the conbercept group received an intravitreal injection （5 μL） once. On days 7 and 14， fundus fluorescein angiography （FFA） was used to evaluate the fluorescein leakage （gray value） of CNV. Hematoxylin-eosin （HE） staining was adopted to observe the histopathological changes of CNV. Western blot was employed to detect the protein expression levels of hypoxia-inducible factor-1<italic>α</italic> （HIF-1<italic>α</italic>） and vascular endothelial growth factor （VEGF） in the retina and choroid. Result:FFA results showed that compared with the normal group， other groups showed increased gray value on days 7 and 14 （<italic>P</italic><0.01）. On day 7， the gray value of the high-dose ART group and the conbercept group decreased compared with that in the model group （<italic>P</italic><0.01）. On day 14， the gray value of the ART groups and the conbercept group decreased in varying degrees compared with that in the model group （<italic>P</italic><0.05， <italic>P</italic><0.01）. HE results showed that compared with the normal group， the model group showed increased thickness of CNV on days 7 and 14 （<italic>P</italic><0.01）. Compared with the model group， the ART groups and the conbercept group displayed decreased thickness of CNV （<italic>P</italic><0.01）. Western blot results revealed that the expression of HIF-1<italic>α</italic> and VEGF in the model group increased in varying degrees on the days 7 and 14 compared with that in the normal group （<italic>P</italic><0.05， <italic>P</italic><0.01）， while compared with the model group， the ART groups and the conbercept group showed decreased expression （<italic>P</italic><0.01）. Conclusion:ART can inhibit experimental CNV by down-regulating the expression of HIF-1<italic>α</italic> and VEGF in the early stage of experimental CNV formation.

Artesunate alleviates renal ischemia-reperfusion injury in rats by inhibiting pyroptosis via NLRP3 inflammasome / 器官移植

Objective To investigate the effect and mechanism of artesunate on renal ischemia-reperfusion injury (IRI) in rat. Methods Twenty-five SD rats were randomly divided into the sham operation group (Sham group), model group (IRI group), low-dose artesunate group (ART-L group), high-dose artesunate group (ART-H group) and NLRP3 inflammasome inhibitor group (INF39 group), with 5 rats in each group. The levels of serum creatinine (Scr), blood urea nitrogen (BUN) and pathological damage of renal tissue were analyzed. The levels of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in serum were quantitatively measured. The expression level of IL-1β in renal tissues was determined by immunohistochemical staining. The expression levels of pyroptosis-related proteins were detected by fluorescent staining and Western blot. Results Compared with the Sham group, the levels of Scr and BUN were higher, the renal tissue injury was aggravated, the expression levels of TNF-α, IL-6 and IL-1βwere higher, and the expression levels of kidney injury molecule (KIM-1), pyroptosis-related protein NOD-like receptor protein 3 (NLRP3), cysteinyl aspartate specific proteinase (Caspase-1), Gasdermin D (GSDMD) and IL-1β proteins were higher in the IRI group. Compared with the IRI group, the levels of Scr and BUN were decreased, the renal tissue injury was mitigated, the expression levels of TNF-α, IL-6 and IL-1β were down-regulated, and the expression levels of KIM-1, NLRP3, Caspase-1, GSDMD and IL-1β proteins were down-regulated in the ART-L, ART-H and INF39 groups. Conclusions Artesunate may inhibit pyroptosis induced by NLRP3 inflammasome, down-regulate the expression levels of pyroptosis -related proteins, reduce the release of inflammatory factors after renal IRI and alleviate renal IRI.

Chemotherapeutic Interaction of Vernonia amygdalina (Delile) Leaf Extract with Artesunate and Amodiaquine in Murine Malaria Model

Aim of the Study: Conventional antimalarial drugs are used concurrently with herbal remediesin malarial endemic developing countries.Vernonia amygdalina is one of such popular herbs used in the treatment of malaria. This study aimed at investigating the antimalarial chemotherapeutic interaction ofVernonia amygdalina (VA) when combined with Amodiaquine (AQ) and/or Artesunate (AS) in a murine Plasmodium berghei malaria model.Methodology:Various doses of aqueous VA leaf extract (100-500 mg/kg/day), AQ (2-10 mg/kg/day) and AS (0.8-4 mg/kg/day) wereadministered orally to P berghei.-infected Swiss albino mice to determine their sub-therapeutic doses. These doses were subsequently used to investigate the chemotherapeutic interactions of VA with AQ and/or AS in both early and establishedmalaria infection test models. The survival of animals with established infections that received different drug/herb treatments were determined using their mean survival time (days) and Kaplan-Meier survival curves (percentage). Using GraphPad Instat (version 3.10) and PrismR(version 5.01) the data obtained were subjected to One-way ANOVA, followed by Student-Newman-Keuls test. P< .05 was considered statistically significant.Results:The sub-therapeutic doses of VA, AQ and AS were found to be 100 mg/kg, 2 mg/kg and 2.4 mg/kg, respectively. The chemosuppressive effect of AQ or AS was significantly increased (p< 0.05) when administered in combination with the VA extract. Similarly, combination of VA extract with AQ or AS resulted in significant (P < .05) parasite clearance when compared to the effects of the herb or the conventional drugs administered separately. The mean survival period of animals with established infection was also significantly enhanced by the VA alone or with AQ(or AS) compared to placebo

Therapeutic effects of artesunate on cytomegalovirus pneumonia in mice / 药学学报

To investigate the therapeutic effect of artesunate on mouse cytomegalovirus pneumonia, the BALB/c-nu mice were infected with murine cytomegalovirus-green fluorescent protein (MCMV-GFP) by nose dropping method. The experimental protocol was approved by the Medical Laboratory Animal Ethics Committee of Guangzhou Medical University. The BALB/c-nu mice were randomly divided into five groups: control group, MCMV pneumonia group, and artesunate (60, 120, and 240 mg·kg-1) groups. The survival rate, weights, and virus loads in lungs among the groups were observed. The degree of histopathologic changes in lungs was assessed directly by hematoxylin-eosin (HE) assay. MCMV-GFP expression was assessed by immunofluorescence. In addition, reverse transcription polymerase chain reaction (RT-PCR) analysis was performed to investigate the content of major immediate early 1 (Mie1) mRNA, and enzyme-linked immunosorbent assay (ELISA) was used to detect the changes of inflammatory factors, interleukin 10 (IL-10), IL-6, and tumor necrosis factor-α (TNF-α). Western blot analysis was used to detect the expression of the changes of nuclear factor-kappa B (NF-κB) signaling pathways in total proteins. Compared with MCMV group, artesunate (120 mg·kg-1) significantly increased body weights of MCMV-infected nude mice over 30 days, and decreased the viral titer in lung homogenate, lung inflammation, and histological severity. Moreover, the administration of artesunate (120 mg·kg-1) could downregulate the expression of phospho-NF-κB (p-NF-κB) p65 in the lungs of mice. The present study suggested that artesunate can protect the immunocompromised mice from MCMV-induced interstitial pneumonia via downregulating NF-κB signaling pathway, thus attenuating inflammation in the lungs.

Study on Preparation and in vitro Anticancer Activity of PFV-modified Paclitaxel/artesunate Co-loaded Targeting Micelles / 中国药房

OBJECTIVE：To prepare cell penetrating peptide PFV-modified paclitaxel （PTX）/artesunate（ART）co-loaded targeting micelles ，and to investigate in vitro anti-tumor activity. METHODS ：According to optimal technology ，PFV-modified PTX/ ART co-loaded targeting micelles were prepared by membrane hydration method ，and were characterized. Using blank micelle as blank control ，sulforhodamine B （SRB）method was used to evaluate the toxicity of PTX micelles ，ART micelles ，PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles to human gastric cancer BGC- 823 cells. The coumarin was used as fluorescent probe replacing PTX to prepare corresponding micelles. Then ，the uptake of BGC- 823 cells to corresponding micelles and targeting effect were observed and determined by flow cytometry and fluorescence microscope. The effects of PTX micelles ， ART micelles ，PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles on the invasion of BGC- 823 cells were investigated by Transwell chamber method. RESULTS ：Average particle size of PFV-modified PTX/ART co-loaded targeting micelles was （51.30±3.95）nm；PDI was 0.19±0.01，and Zeta potential was （0.21±0.02）mV. The encapsulation efficiency of PTX and ART were higher than 90％. The shape of micelles were spherical. The blank micelles had no obvious toxicity to BGC-823 cells. The IC 50 value of PTX micelles ，PTX/ART micelles and PFV-modified PTX/ART co-loaded targeting micelles to BGC-823 cells were （3.09±0.22），（1.93±0.24），（1.11±0.15）μmol/L，respectively. The distribution amount of different micelles in BGC- 823 cell nucleus in the descending order were PFV-modified coumarin/ART micelles ＞coumarin/ART micelles ＞coumarin micelles＞blank control. The order of inhibitory effect was PFV-modified PTX/ART co-loaded targeting micelles ＞PTX/ART micelles＞ART micelles ＞PTX micelles ＞blank control. CONCLUSIONS： Prepared PFV-modified PTX/ART No.81874347） co-loaded targeting micelles are in line with the quality of 1915286446@qq.com Chinese Pharmacopoeia . It shows strong cytotoxicity to BGC-823 cells，can improve the drug targeting and the cell uptake，and inhibit the inv asion and metastasis of BGC- 823 cells.

Inhibitory effect of artesunate on the growth of breast cancer cells and its mechanism / 解放军医学杂志

Objective To investigate the function of artesunate on the growth of breast cancer cell line MDA-MB-231 and its potential mechanism. Methods CCK-8 assays were used to measure growth inhibition in breast cancer cells (MDA-MB-231) in the presence of artesunate, the IC50 was calculated. Based on the IC50 values, various artesunate concentrations (0, 25, 50, 100 μg/ml) were used to experiments in this paper. 0 μg/ml artesunate group served as the control group, using normal saline instead of the artesunate. Apoptosis and mitochondrial membrane potential were analyzed by flow cytometry, after the treatment of various artesunate concentrations (0, 25, 50, 100 μg/ml) for 24 hours. Migration and invasion of the MDA-MB-231 cells were evaluated using wound healing and Transwell assays, respectively, after the treatment of various artesunate concentrations (0, 25 μg/ml) for 24 hours. Results Artesunate inhibited the growth of MDA-MB-231 cells in a dose-dependent manner, with IC50 values for MDA-MB-231 of 54.24 μg/ml. Compared to mock-treated cells, artesunate significantly increased apoptosis, while the mitochondrial membrane potential was significantly decreased after 24 h of exposure to different concentrations of artesunate (P<0.01), in a dose-dependent manner. The cell migration and invasion abilities were also lower in the 25 μg/ml artesunate treated cells than these abilities in the mock-treated cells. Conclusions Artesunate could inhibit the growth of MDA-MB-231 breast cancer cells and inhibit its invasion and migration ability by inducing apoptosis and decreasing mitochondrial membrane potential.

Artesunate relieves subchondral bone osteoclast-mediated osteoarthritis pain / 中国组织工程研究

BACKGROUND: Studies have shown that osteoclasts induce aberrant in growth of sensory nerves into the subchondral bone by secreting netrin-1, resulting in a reduced pain threshold in an osteoarthritis animal model. Therefore, we assume that inhibition of osteoclasts can alleviate sensory nerve-mediated pain symptoms. OBJECTIVE: To investigate whether artesunate inhibits subchondral bone osteoclasts and reduces sensory nerve-mediated pain, providing experimental data for the treatment of osteoarthritis pain using artesunate. METHODS: C57BL/6J male mice were randomly assigned to a sham operation group, a placebo group and an artesunate group, with 10 mice per group. The mice in the sham operation group were only subjected to right knee capsulotomy with no damage to the other structures. Moreover, there was no intervention after operation. In the other two groups, the mice received an anterior cruciate ligament transection of the right knee to establish the osteoarthritis model, followed by treatment with artesunate (artesunate group, 100 mg/kg per day) or equivalent volume of 5% NaHCO3 (placebo group) via intraperitoneal injection. Fourteen days after surgery, the footprint trial was performed, and the levels of tartrate-resistant acid phosphatase 5b (TRAcP5b), cathepsin K and carboxy-terminal telopeptide of type I collagen (CTX-I) in the peripheral blood were detected using ELISA. The knee joint specimens of each group were subjected to Safranin O-Fast Green staining, histological scoring, tartrate-resistant acid phosphatase (TRAP) staining and immunohistochemical staining with netrin-1 and calcitonin gene-related peptide (CGRP). RESULTS AND CONCLUSION: The percentage of ipsilateral contact area of the right hindpaw in the footprint trial was significantly higher in the sham operation group and artesunate group than the placebo group (P 0.05). No significant differences were observed in the serum levels of TRAcP5b, cathepsin K and CTX-I between the groups (P > 0.05). Based on the Safranin O-Fast Green staining, the cartilage histology score was significantly lower in the sham operation group and the artesunate group than the placebo group (P 0.05). TRAP staining indicated that compared with the placebo group, the Trap+ osteoclasts were significantly lower in the sham operation group and the artesunate group (P 0.05). Compared with the placebo group, the netrin-1+ and CGRP+ sensory nerves in the subchondral bone were significantly decreased in the sham operation group and the artesunate group (P 0.05). Our findings from this study indicate that artesunate improves sensory nerve-mediated pain by inhibiting netrin-1 secreted by subchondral bone osteoclasts, and has therapeutic potential to alleviate osteoarthritis pain.