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1.
Journal of Preventive Medicine ; (12): 253-257, 2022.
Article in Chinese | WPRIM | ID: wpr-920762

ABSTRACT

Abstract@#As the largest human microecosystem, intestinal microorganisms participate in human material and energy metabolisms and pose a significant impact on human health. Diabetes mellitus is likely to cause imbalance of abundance and component alterations in intestinal microorganisms, and reduce the diversity and balance, leading to intestinal microflora dysregulation. It has been shown that intestinal microflora dysregulation may promote diabetes development and progression through the reduction of intestinal microbial metabolites, inflammatory reaction and insulin resistance. This review summarizes the involvement of intestinal microorganisms in the pathogenesis of diabetes through metabolites including short-chain fatty acid, bile acid and lipopolysaccharide, and describes the current status of intestinal microorganisms-mediated treatments for diabetes, so as to provide the theoretical basis for the researches on diabetes and intestinal microorganisms.

2.
Acta Pharmaceutica Sinica B ; (6): 2129-2149, 2022.
Article in English | WPRIM | ID: wpr-929399

ABSTRACT

Cardiometabolic disease (CMD), characterized with metabolic disorder triggered cardiovascular events, is a leading cause of death and disability. Metabolic disorders trigger chronic low-grade inflammation, and actually, a new concept of metaflammation has been proposed to define the state of metabolism connected with immunological adaptations. Amongst the continuously increased list of systemic metabolites in regulation of immune system, bile acids (BAs) represent a distinct class of metabolites implicated in the whole process of CMD development because of its multifaceted roles in shaping systemic immunometabolism. BAs can directly modulate the immune system by either boosting or inhibiting inflammatory responses via diverse mechanisms. Moreover, BAs are key determinants in maintaining the dynamic communication between the host and microbiota. Importantly, BAs via targeting Farnesoid X receptor (FXR) and diverse other nuclear receptors play key roles in regulating metabolic homeostasis of lipids, glucose, and amino acids. Moreover, BAs axis per se is susceptible to inflammatory and metabolic intervention, and thereby BAs axis may constitute a reciprocal regulatory loop in metaflammation. We thus propose that BAs axis represents a core coordinator in integrating systemic immunometabolism implicated in the process of CMD. We provide an updated summary and an intensive discussion about how BAs shape both the innate and adaptive immune system, and how BAs axis function as a core coordinator in integrating metabolic disorder to chronic inflammation in conditions of CMD.

3.
Acta Pharmaceutica Sinica B ; (6): 1447-1459, 2022.
Article in English | WPRIM | ID: wpr-929362

ABSTRACT

Cancer remains one of the leading causes of death globally and metastasis always leads to treatment failure. Here, we develop a versatile hydrogel loading photothermal agents, chemotherapeutics, and immune-adjuvants to eradicate orthotopic tumors and inhibit metastasis by combinational therapy. Hydrogel networks were synthesized via the thiol-Michael addition of polydopamine (PDA) with thiolated hyaluronic acid. PDA acted as a cross-linking agent and endowed the hydrogel with excellent photothermal property. Meanwhile, a chemotherapeutic agent, doxorubicin (DOX), was loaded in the hydrogel via π‒π stacking with PDA and an immune-adjuvant, CpG-ODN, was loaded via electrostatic interaction. The release of DOX from the hydrogel was initially slow but accelerated due to near infrared light irradiation. The hydrogels showed remarkably synergistic effect against 4T1 cancer cells and stimulated plenty of cytokines secreting from RAW264.7 cells. Moreover, the hydrogels eradicated orthotopic murine breast cancer xenografts and strongly inhibited metastasis after intratumoral injection and light irradiation. The high anticancer efficiency of this chemo-photothermal immunotherapy resulted from the strong synergistic effect of the versatile hydrogels, including the evoked host immune response. The combinational strategy of chemo-photothermal immunotherapy is promising for highly effective treatment of breast cancer.

4.
Acta Pharmaceutica Sinica B ; (6): 1198-1212, 2022.
Article in English | WPRIM | ID: wpr-929355

ABSTRACT

Pulmonary endothelial barrier dysfunction is a hallmark of clinical pulmonary edema and contributes to the development of acute lung injury (ALI). Here we reported that ruscogenin (RUS), an effective steroidal sapogenin of Radix Ophiopogon japonicus, attenuated lipopolysaccharides (LPS)-induced pulmonary endothelial barrier disruption through mediating non-muscle myosin heavy chain IIA (NMMHC IIA)‒Toll-like receptor 4 (TLR4) interactions. By in vivo and in vitro experiments, we observed that RUS administration significantly ameliorated LPS-triggered pulmonary endothelial barrier dysfunction and ALI. Moreover, we identified that RUS directly targeted NMMHC IIA on its N-terminal and head domain by serial affinity chromatography, molecular docking, biolayer interferometry, and microscale thermophoresis analyses. Downregulation of endothelial NMMHC IIA expression in vivo and in vitro abolished the protective effect of RUS. It was also observed that NMMHC IIA was dissociated from TLR4 and then activating TLR4 downstream Src/vascular endothelial cadherin (VE-cadherin) signaling in pulmonary vascular endothelial cells after LPS treatment, which could be restored by RUS. Collectively, these findings provide pharmacological evidence showing that RUS attenuates LPS-induced pulmonary endothelial barrier dysfunction by inhibiting TLR4/Src/VE-cadherin pathway through targeting NMMHC IIA and mediating NMMHC IIA‒TLR4 interactions.

5.
Acta Pharmaceutica Sinica B ; (6): 511-531, 2022.
Article in English | WPRIM | ID: wpr-929312

ABSTRACT

Aging is by far the most prominent risk factor for Alzheimer's disease (AD), and both aging and AD are associated with apparent metabolic alterations. As developing effective therapeutic interventions to treat AD is clearly in urgent need, the impact of modulating whole-body and intracellular metabolism in preclinical models and in human patients, on disease pathogenesis, have been explored. There is also an increasing awareness of differential risk and potential targeting strategies related to biological sex, microbiome, and circadian regulation. As a major part of intracellular metabolism, mitochondrial bioenergetics, mitochondrial quality-control mechanisms, and mitochondria-linked inflammatory responses have been considered for AD therapeutic interventions. This review summarizes and highlights these efforts.

6.
Article in English | WPRIM | ID: wpr-929264

ABSTRACT

Abelmoschus manihot (L.) Medik. (A. manihot) is a traditional Chinese herbal medicine with a variety of pharmacological properties. It was first recorded in Jiayou Materia Medica dating back to the Song dynasty to eliminate urinary tract irritation by clearing away heat and diuretic effect. However, its pharmacological action on urinary tract infections has not been investigated. The present study aims to evaluate the anti-inflammatory activity of A. manihot on a mouse model of lipopolysaccharide (LPS)-induced cystitis. The results showed that A. manihot decreased white blood cell (WBC) count in urine sediments of the cystitis mice, alleviated bladder congestion, edema, as well as histopathological damage, reduced the expression levels of tumor necrosis factor-α, interleukin-6, and interleukin-1β simultaneously. Moreover, A. manihot administration significantly downregulated the expression levels of TLR4, MYD88, IκBα, p-IκBα, NF-κB p65, and p-NF-κB p65 in LPS-induced cystitis mice. These findings demonstrated the protective effect of A. manihot against LPS-induced cystitis, which is attributed to its anti-inflammatory profile by suppressing TLR4/MYD88/NF-κB pathways. Our results suggest that A. manihot could be a potential candidate for cystitis treatment.


Subject(s)
Abelmoschus/metabolism , Animals , Anti-Inflammatory Agents/therapeutic use , Cystitis , Female , Humans , Lipopolysaccharides/pharmacology , Male , Mice , Myeloid Differentiation Factor 88/metabolism , NF-KappaB Inhibitor alpha/metabolism , NF-kappa B/metabolism , Signal Transduction , Toll-Like Receptor 4/metabolism
7.
Article in English | WPRIM | ID: wpr-928969

ABSTRACT

OBJECTIVES@#Chlorogenic acid has various physiological activities such as antibacterial, anti-inflammatory, and antiviral activities. Studies have shown that chlorogenic acid can alleviate the inflammatory response of mice with acute lung injury (ALI), but the specific mechanism is still unclear. This study aims to investigate whether chlorogenic acid attenuates lipopolysaccharide (LPS)-induced ALI in mice by regulating the microRNA-223 (miR-223)/nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) axis.@*METHODS@#SPF grade BALBc male mice were randomly divided into a control group, a model group, a chlorogenic acid group, a chlorogenic acid+miR-223 negative control (miR-223 NC) group, and a chlorogenic acid+miR-223 inhibitor (miR-223 antagomir) group, 10 mice in each group. Except the control group, the other groups were instilled with 4 mg/kg LPS through the airway to establish the ALI mouse model. After the modeling, the mice in the chlorogenic acid group were continuously given chlorogenic acid (100 mg/kg) by gavage for 7 d. The chlorogenic acid+miR-223 NC group and the chlorogenic acid+miR-223 antagomir group were given 100 mg/kg chlorogenic acid by gavage every day, and then were injected with 10 μL of miR-223 NC (0.5 nmol/μL) and miR-223 antagomir (0.5 nmol/μL) respectively for 7 consecutive days.The control group and the model group were replaced with normal saline. The lung tissues of mice were taken to measure the ratios of lung wet to dry weight (W/D). The bronchoalveolar lavage fluid of mice was collected to measure the levels of TNF-α, IL-6, and IL-1β by ELISA kit and to count the number of eosinophils (EOS), lymphocytes, neutrophils under light microscope. After HE staining, the pathological changes of lung tissues were observed and lung injury was scored. qRT-PCR method were used to determine the expression levels of miR-223 in lung tissues. Western blotting was used to determine the expression levels of NLRP3 protein in mouse lung tissues. Luciferase reporter assay was used to analyze the targeting relationship of miR-223 to NLRP3.@*RESULTS@#Compared with the control group, the lung W/D value, the lung injury score and the level of inflammatory factors in the bronchoalveolar lavage fluid were significantly increased in the model group (all P<0.05); the infiltration of inflammatory cells in the lung tissue was severe; the alveolar space was significantly increased; the alveolar wall was significantly thickened; the number of EOS, lymphocytes, and neutrophils in the bronchoalveolar lavage fluid was significantly increased (all P<0.05); the expression levels of miR-223 in lung tissue were significantly decreased (P<0.05); and the protein expression levels of NLRP3 were significantly increased (P<0.05). Compared with the model group, the W/D value of lungs, lung injury score, and levels of inflammatory factors in bronchoalveolar lavage fluid were significantly decreased in the chlorogenic acid group, the chlorogenic acid+miR-223 NC group, and the chlorogenic acid+miR-223 antagomir group (all P<0.05); lung tissues damage was alleviated; the numbers of EOS, lymphocytes, and neutrophils in bronchoalveolar lavage fluid were significantly decreased (all P<0.05); the expression levels of miR-223 in lung tissues were significantly increased (P<0.05); and the expression levels of NLRP3 protein were significantly decreased (P<0.05). Compared with the chlorogenic acid group, the lung W/D value, lung injury score, and inflammatory factor levels in the bronchoalveolar lavage fluid were significantly increased in the chlorogenic acid+miR-223 antagomir group (all P<0.05); lung tissue damage was aggravated; the number of EOS, lymphocytes and neutrophils in bronchoalveolar lavage fluid significantly increased (all P<0.05); the expression levels of miR-223 in lung tissues were significantly decreased (P<0.05); and the expression levels of NLRP3 protein were significantly increased (P<0.05). The results of luciferase reporter assay showed that miR-223 had a targeting relationship with NLRP3.@*CONCLUSIONS@#Chlorogenic acid may increase the level of miR-223, target the inhibition of NLRP3 expression, reduce LPS-induced inflammatory response in ALI mice, and alleviate pathological damage of lung tissues.


Subject(s)
Acute Lung Injury/genetics , Animals , Antagomirs/metabolism , Bronchoalveolar Lavage Fluid , Chlorogenic Acid/metabolism , Lipopolysaccharides/adverse effects , Lung/pathology , Male , Mice , MicroRNAs/metabolism , NLR Family, Pyrin Domain-Containing 3 Protein/genetics
8.
Article in English | WPRIM | ID: wpr-928241

ABSTRACT

Objective To examine the neuroanatomical substrates underlying the effects of minocycline in alleviating lipopolysaccharide (LPS)-induced neuroinflammation. Methods Forty C57BL/6 male mice were randomly and equally divided into eight groups. Over three conse-cutive days, saline was administered to four groups of mice and minocycline to the other four groups. Immediately after the administration of saline or minocycline on the third day, two groups of mice were additionally injected with saline and the other two groups were injected with LPS. Six or 24 hours after the last injection, mice were sacrificed and the brains were removed. Immunohistochemical staining across the whole brain was performed to detect microglia activation via Iba1 and neuronal activation via c-Fos. Morphology of microglia and the number of c-Fo-positive neurons were analyzed by Image-Pro Premier 3D. One-way ANOVA and Fisher's least-significant differences were employed for statistical analyses. Results Minocycline alleviated LPS-induced neuroinflammation as evidenced by reduced activation of microglia in multiple brain regions, including the shell part of the nucleus accumbens (Acbs), paraventricular nucleus (PVN) of the hypothalamus, central nucleus of the amygdala (CeA), locus coeruleus (LC), and nucleus tractus solitarius (NTS). Minocycline significantly increased the number of c-Fo-positive neurons in NTS and area postrema (AP) after LPS treatment. Furthermore, in NTS-associated brain areas, including LC, lateral parabrachial nucleus (LPB), periaqueductal gray (PAG), dorsal raphe nucleus (DR), amygdala, PVN, and bed nucleus of the stria terminali (BNST), minocycline also significantly increased the number of c-Fo-positive neurons after LPS administration. Conclusion Minocycline alleviates LPS-induced neuroinflammation in multiple brain regions, possibly due to increased activation of neurons in the NTS-associated network.


Subject(s)
Animals , Female , Lipopolysaccharides/toxicity , Male , Mice , Mice, Inbred C57BL , Minocycline/pharmacology , Neuroinflammatory Diseases , Solitary Nucleus
9.
China Pharmacy ; (12): 1436-1441, 2022.
Article in Chinese | WPRIM | ID: wpr-927189

ABSTRACT

OBJECTIVE To investigate the effect of arctiin (ARC)relieving lipopolysaccharide (LPS)induced inflammatory injury of human nasopharyngeal epithelial cells NP- 69. METHODS The effects of 24 h treatment of 0.000 1,0.001,0.01,0.1, 1.0,10 μmol/L ARC on the proliferation of NP-69 were determined by MTS method. After 0.01,0.1,and 1.0 μmol/L ARC was applied to NP- 69 for 24 h and NP- 69 was pre-treated with 0.01,0.1 and 1.0 μmol/L ARC for 24 h,and then stimulated with 1.0 μg/mL LPS for 24 h,scratch tests were used to detect cell migration in both experiments. LPS stimulated NP- 69 to establish an inflammation injury model. The levels of nitric oxide (NO),tumor necrosis factor α(TNF-α)interleukin-6(IL-6),and IL- 1β in cell supernatants were detected ,and mRNA and protein expression of zonula oecludens protein 1(ZO-1),β-defensin 3(BD3), Janus kinase 1 (JAK1),signal transducer and activator of transcription 3 (STAT3) in cell supernatant were also detected. RESULTS Compared with normal group ,0.000 1,0.001,0.01,0.1,1.0,10 μmol/L ARC had no effect on the proliferation of NP-69 after 24 h treatment (P>0.05). ARC (0.1,1.0 μmol/L)could significantly promote the rate of cell migration (P<0.05). For the inflammatory injure of NP- 69 cells stimulated by LPS ,ARC(1.0 μmol/L)could significantly reduce the release of NO , TNF-α and IL-6(P<0.05),significantly increased mRNA and protein expression of ZO- 1 and BD 3 but decreased mRNA and protein expression of STAT 3(P<0.01 or P<0.05). CONCLUSIONS ARC has the effect of reducing the inflammatory injury of NP-69 cells induced by LPS ,promoting the physical and immune defense ability of the nasal mucosa epithelial barrierunder inflammatory environment. The mechanism of action may be related to inhibiting IL- 6/JAK1/STAT3 signaling pathway.

10.
Braz. J. Pharm. Sci. (Online) ; 58: e181092, 2022. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1374550

ABSTRACT

Abstract The present study was designed to examine the effects of atorvastatin on vascular inflammatory responses in human coronary artery endothelial cells(HCAECs), when challenged by lipopolysaccharide (LPS), a Toll-like receptor-4 (TLR4) ligand. HCAECs were pretreated with atorvastatin and induced by LPS. The expression of TLR4, interleukin -6(IL-6), monocyte chemoattractant protein 1(MCP-1), vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecular-1(ICAM-1), nuclear factor-κB (NF-κB) and p38 mitogen activated protein kinase(p38 MAPK) were evaluated using Real-time polymerase chain reaction, cytokine ELISA assay and Western blotting. The results showed that pretreatment with atorvastatin down-regulated the expression of TLR4 in LPS-activated HCAECs. Atorvastatin also attenuated the LPS-induced expression of interleukin IL-6 and MCP-1, at both the transcription and translation level in HCAECs. LPS-induced endothelial cell adhesion molecules, ICAM-1 and VCAM-1 expression were also reduced by pretreatment with atorvastatin. Furthermore, atorvastatin efficiently suppressed LPS-induced phosphorylation of NF-κB and p38 MAPK in HCAECs. These findings show that atorvastatin suppresses endothelial cell inflammation, suggesting that atorvastatin may be suitable for development as a therapeutic agent for inflammatory cardiovascular disease.

11.
Article in Chinese | WPRIM | ID: wpr-875673

ABSTRACT

Objective To study the effect of nicotinamide mononucleotide (NMN) on the mortality of the lipopoly-saccharide (LPS)-induced endotoxic shock mouse model. Methods 10-week-old C57BL/6J male mice were randomly divided into groups, and were injected intraperitoneally (i.p.) with LPS (10 mg/kg) to induce endotoxic shock models. NMN was i.p. injected in three ways: (1) 0.5 h after modeling, doses of 10, 30, 100 and 300 mg/kg; (2) 0.5 h before modeling, doses of 30, 100, 300 and 600 mg/kg; or (3) 0.5 and 12 h after modeling, dose of 300 mg/kg each time. The death times of each group were recorded, and the survival curves were drawn. Results Compared with the solvent control group, NMN at different doses given 0.5 h after or before modeling didn’t improve the survival rate or delay the death time of endotoxic shock mice; But when given at 0.5 and 12 h 300 mg/kg after modeling, NMN accelerated the death of mice and increased the mortality of mice. NMN products by two manufacturers showed similar effects. Conclusion NMN has no therapeutic effect on LPS-induced endotoxic shock, and repeated administration of NMN after endotoxic shock will increase the mortality.

12.
International Eye Science ; (12): 411-416, 2021.
Article in Chinese | WPRIM | ID: wpr-873434

ABSTRACT

@#AIM: To explore the effect of lycium barbarum polysaccharides(LBP)on inflammatory response of human retinal pigment epithelial cells(ARPE-19)induced by lipopolysaccharide(LPS)and its possible signal pathway.<p>METHODS: ARPE-19 cells were stimulated by LPS <i>in vitro</i> to construct the inflammatory injury cell model. Primarily, the cells were divided into five groups randomly. The blank group was cultured in complete medium, and the LPS group was stimulated with complete medium containing 10μg/mL LPS for 24h. The low, medium and high concentration LBP groups were incubated with complete medium importing 0.1, 0.5 and 1mg/mL LBP for 24h separately, and then stimulated with complete medium containing 10μg/mL LPS for 24h. We used the CCK-8 method to observe the cell survival rate, real-time fluorescent quantitative PCR to detect the mRNA expression of inflammatory factors and Western blot to test the changes of phosphorylated protein within the signaling pathway of NF-κB/MAPK.<p>RESULTS: Compared with normal cells, the survival rate of ARPE-19 cells was decreased after the LPS stimulation. With the increase of exogenous LBP concentration, the survival rate of ARPE-19 cell was gradually increased, while the inflammatory factors expression of cytokines IL-1β, IL-6 and MCP-1 were reduced accompany with the phosphorylated proteins(p-p65, P-IκBα, p-JNK, p-ERK and p-p38)of NF-κB/MAPK signaling pathway were decreased.<p>CONCLUSION: LBP prevents LPS-induced inflammatory response of ARPE-19 by inhibiting the intracellular inflammatory factors and the phosphorylation of the related protein within NF-κB/MAPK signaling pathway.

13.
International Eye Science ; (12): 217-221, 2021.
Article in Chinese | WPRIM | ID: wpr-862414

ABSTRACT

@#AIM: To further explore effective drugs for dry eye treatment by isolating and culturing lacrimal gland epithelial cells<i> in vitro</i>, establishing a dry eye cell model and analyzing relevant inflammatory factors. <p>METHODS: Rabbit lacrimal gland epithelial cells were <i>in vitro</i> isolated and cultured, and the activity and purity of primary cells were identified by cell proliferation experiment and immunofluorescence experiment. In addition, 0.5 times IC<sub>50</sub> of lipopolysaccharide LPS and TNF-α were used respectively to stimulate rabbit lacrimal gland epithelial cells and then establish two dry eye cell models. Finally, through cell proliferation experiment, ELISA and flow cytometry, the biological characteristics of these two dry eye cell models were compared. <p>RESULTS:After 12h of culture, the primary cells of lacrimal gland epithelial cells basically adhered to the wall of culture bottles; and 48h later, the cells stretched and almost each of them presented a shape of a long triangle. The activity of primary cells of lacrimal gland epithelium was 92%, and the positive rate of marker Pan-rkeratin was more than 90%, which accorded with the experimental requirements. The IC<sub>50</sub> of LPS and TNF-α are 20μg/mL and 4.996ng/mL respectively. After 12h of intervention with LPS(10μg/mL)and TNF-α(2.5ng/mL), the cell activity of the two groups was significantly lower than that of control group(<i>P</i><0.01); compared between these two groups, the apoptosis rate of TNF-α group is higher than that of LPS group(<i>P</i><0.01). The levers of IL-1β and IL-6 in the cell supernatants of the two groups were significantly higher than those of the control group(<i>P</i><0.01); compared between the two groups, IL-1β and IL-6 in TNF-α group were significantly higher than those in LPS group(<i>P</i><0.01). It was suggested that TNF-α was superior to LPS in simulating inflammatory response of dry eye. <p>CONCLUSION: This study successfully established a relatively simple and rapid rabbit dry eye cell model with high cell purity and stability, which provided a more stable <i>in vitro</i> experimental model for the basic research on the function of rabbit lacrimal gland epithelial cells and dry eye.

14.
Article in English | WPRIM | ID: wpr-862226

ABSTRACT

@#BACKGROUND: Acute respiratory distress syndrome (ARDS) causes substantial mortalities. Alveolar epithelium is one of the main sites of cell injuries in ARDS. As an important kind of microRNAs (miRNAs), microRNA-145 (miR-145) has been studied in various diseases, while its role in ARDS has not been investigated. METHODS: Lipopolysaccharide (LPS) was intratracheally instilled to establish a rat ARDS model. Cytokines from bronchoalveolar lavage fluid (BALF) were measured using rat tumor necrosis factor-α and interleukin-6 enzyme-linked immunosorbent assay kits (R&D Systems), and the pathological structures were evaluated using hematoxylin and eosin (H&E) staining and transmission electron microscope; the lung miR-145 messenger RNA (mRNA) was detected using quantitative polymerase chain reaction. Bioinformatics focused on the target genes and possible pathways of gene regulation. RESULTS: A rat model of LPS-induced ARDS was successfully established. The miR-145 was down-regulated in the LPS-induced ARDS lung, and mitochondrial dysfunction was observed in alveolar epithelial cells, most obviously at 72 hours after LPS. TargetScan and miRDB databases were used to predict the target genes of miR-145. A total of 428 overlapping genes were identified, seven genes were associated with mitochondrial function, and Ogt, Camk2d, Slc8a3, and Slc25a25 were verified. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were enriched in the mitogen-activated protein kinase (MAPK) signaling pathway, and Gene Ontology (GO) biological process was mainly enriched in signal transduction and transcription regulation. CONCLUSIONS: The miR-145 is down-regulated in LPS-induced ARDS, and affects its downstream genes targeting mitochondrial functions.

15.
Article in Chinese | WPRIM | ID: wpr-907719

ABSTRACT

Objective:To investigate the role of KLF4 in lipopolysaccharide induced cardiomyocyte injury.Methods:Primary rat cardiomyocytes were isolated and cultured, and randomly divided into 5 groups: control group, negative control (NC), LPS group, KLF4 overexpression group, KLF4 overexpression+LPS group. MTT method was used to detect cell activity, ROS, SOD 2, GPX and MDA were detected by kit, TNFa, IL-1 β and IL-6 were detected by ELISA. TUNEL staining was used to detect apoptosis. The protein levels of TLR4 and Nrf2 were detected by Western blot.Results:The expression of KLF4 in cardiomyocytes was significantly higher than that in the NC group ( P<0.001). The cell activity of LPS group was significantly lower than that of NC group ( P < 0.001), and that of KLF4 overexpression +LPS group was higher than that of LPS group ( P<0.001). The levels of TNFa, IL-1 β and IL-6 in LPS group were significantly higher than those in the NC group ( P<0.0001), and the levels of TNFa, IL-1 β and IL-6 in KLF4 overexpression +LPS group were lower than those in LPS group ( P<0.0001). The levels of ROS and MDA in LPS group were significantly higher than those in the control group, while the activities of SOD2 and GPX were lower than those in the NC group ( P<0.0001); the levels of ROS and MDA in KLF4 overexpression +LPS group were lower than those in LPS group, while the activities of SOD2 and GPX were higher than those in LPS group ( P<0.0001). The number of apoptosis in LPS group was significantly higher than that in the NC group, and that in KLF4 overexpression +LPS group was lower than that in LPS group ( P< 0.001). The level of TLR4 wan higher and Nrf2 protein in the nucleus of LPS group was lower than that of the NC group. The level of TLR4 was lower and Nrf2 protein in the nucleus of KLF4 overexpression+LPS group was significantly higher than that of LPS group ( P < 0.001). Conclusions:KLF4 can alleviate LPS induced cardiomyocyte injury by regulating TLR4 and NRF2 signals.

16.
Article in Chinese | WPRIM | ID: wpr-906473

ABSTRACT

Objective:To investigate the effect of Huayu Jiedu prescription (HYJDP) on gut microbiota and fecal metabolites in mice with endometriosis. Method:Normal female C57BL/6J mice were divided into normal control group (CO), endometriosis group (EM) and Chinese medicine Huayu Jiedu decocotion group (CM). CO and EM groups received normal saline and CM group received HYJDP by intragastric administration. Untargeted metabolomics method was used to detect metabolites in fecal supernatant of mice, and receiver operating characteristic (ROC) analysis was used to screen the differential metabolites, 16S rRNA high-throughput sequencing was used to detect the gut microbiota, and Spearman correlation coefficient was used to represent the degree of correlation between differential metabolites and intestinal flora. Lipopolysaccharides (LPSs) in intestinal wall tissue, serum and peritoneal lavage fluid were detected by enzyme-linked immunosorbent assay (ELISA). The expression of Vimentin and E-cadherin in ectopic lesions was detected by immunohistochemistry. Result:HYJDP alleviated the disorders of fecal metabolites and gut microbiota in EMS mice, especially with the recovered levels of homoveratric acid, melilotoside C and physapubescin in fecal supernatant. In the comparison of these three factors between EM group and CO group as well as between EM group and CM group, the variable important in projection (VIP) value was both above 2, and AUC in ROC analysis was both >0.9. As compared with EM group, HYJDP restored the abundance of species such as <italic>Lachnospiraceae_NK4A136_group</italic>, <italic>Lactobacillus</italic> and <italic>Blautia </italic>(<italic>P</italic><0.05). In addition, the level of LPS in peritoneal fluid supernatant of EM group was significantly higher than that of CO group (<italic>P</italic><0.05) and CM group (<italic>P</italic><0.05). The protein expression of vimentin and E-cadherin in endometriosis decreased significantly (<italic>P</italic><0.05). Conclusion:HYJDP which can improve the intestinal environment and reduce the level of LPS in mice with endometriosis, is an effective drug for the treatment of endometriosis.

17.
Article in Chinese | WPRIM | ID: wpr-906457

ABSTRACT

Objective:To explore the effect of Bushen Huatan prescription on serum lipopolysaccharide (LPS) and Toll-like receptor 4 (TLR4)/ myeloid cell differentiation protein 88 (MyD88)/nuclear transcription factor-<italic>κ</italic>B (NF-<italic>κ</italic>B) signaling pathway in rats with ovariectomy-induced osteoporosis. Method:Sixty SPF 6-month-old female rats were randomly divided into sham operation group, model group, estradiol valerate group and Bushen Huatan prescription low, medium and high dose groups.One week after modeling by bilateral ovariectomy, 8 rats in each group were selected to receive intragastric administration.The estradiol valerate group was given 0.184 mg·kg<sup>-1</sup> by gavage, and Bushen Huatan prescription low, middle and high dose groups were given 4.7, 9.4 and 18.8 g·kg<sup>-1</sup> by gavage, sham operation group and model group were given 0.9% saline 4 mL by gavage respectively.After 12 weeks of intervention, the rats were sacrificed for detection.Serum LPS was detected by enzyme linked immunosorbent assay (ELISA), while protein expressions of TLR4, MyD88 and phosphorylated (p)-NF-<italic>κ</italic>B p65 in bone tissue were detected by Western blot, and the mRNA expressions of TLR4, MyD88, NF-<italic>κ</italic>B p65, IL-1<italic>β</italic>, and IL-6 in bone tissue were detected by quantitative real-time polymerase chain reaction(PCR). Result:Compared with sham operation group, the serum LPS level as well as protein expression of TLR4, MyD88, p-NF-<italic>κ</italic>B p65 and mRNA expression of TLR4, MyD88, NF-<italic>κ</italic>B p65, IL-1<italic>β</italic>, and IL-6 significantly increased in model group(<italic>P</italic><0.05).Compared with the model group, serum LPS level, protein expression of TLR4, MyD88, and p-NF-<italic>κ</italic>B p65, mRNA levels of TLR4, MyD88, and NF-<italic>κ</italic>B p65 in bone tissues as well as downstream inflammatory factors IL-1<italic>β</italic>, IL-6 mRNA expression decreased to different degrees in estradiol valerate group and Bushen Huatan prescription high dose group(<italic>P</italic><0.05). Conclusion:Bushen Huatan prescription can reduce serum LPS content, regulate mRNA and protein expression of TLR4, MyD88, NF-<italic>κ</italic>B p65 and p-NF-<italic>κ</italic>B p65 in TLR4/MyD88/NF-<italic>κ</italic>B pathway, and down-regulate mRNA levels of IL-1<italic>β</italic> and IL-6 in bone tissues to improve bone microstructure and inhibit the development of postmenopausal osteoporosis (PMOP).

18.
Article in Chinese | WPRIM | ID: wpr-906401

ABSTRACT

Objective:To compare the contents of adenosine, gastrodin, <italic>p</italic>-hydroxybenzyl alcohol, <italic>p</italic>-hydroxybenzaldehyde, parisinin B and parisinin A in Chijian (the aerial part of <italic>Gastrodia elata</italic>) and Gastrodiae Rhizoma, and compare their effects on immune function and intestinal microflora, evaluating whether it is necessary to study and develop Chijian. Method:The contents of these six constituents were determined by ultra performance liquid chromatography (UPLC), the mobile phase was 0.1% formic acid aqueous solution (A)-acetonitrile (B) for gradient elution (0-4 min, 0.5%B; 4-5 min, 0.5%-2%B; 5-10 min, 2%-15%B; 10-12 min, 15%-20%B; 12-15 min, 20%-95%B; 15-17 min, 95%B; 17-17.5 min, 95%-0.5%B; 17.5-20 min, 0.5%B), the flow rate was 0.5 mL·min<sup>-1</sup>, the detection wavelength was 270 nm. The difference of pharmacological activity of water extracts of Chijian and Gastrodiae Rhizoma was compared, the clearance index, corrected clearance index and peripheral blood were measured in mice model with low immune function induced by cyclophosphamide, B lymphocyte proliferation was determined by lymphocyte transformation test <italic>in vitro</italic>, intestinal microflora was analyzed by 16S rDNA technology and bioinformatics was conducted. Result:The total contents of these six components in powder and ethanol extract of Chijian were higher than that of Gastrodiae Rhizoma, but the total contents of these six components in their water extract were similar, and the total contents of gastrodin and <italic>p</italic>-hydroxybenzyl alcohol met the requirements of the 2020 edition of <italic>Chinese Pharmacopoeia</italic>. Compared with the blank group, the clearance index of immunocompromised mice was significantly increased in the middle-dose (10 g·kg<sup>-1</sup>) group of Chijian water extract, middle- and low-dose (10, 5 g·kg<sup>-1</sup>) groups of Gastrodiae Rhizoma water extract (<italic>P</italic><0.05), the levels of erythrocyte and hematocrit in peripheral blood were significantly increased in the high-dose (20 g·kg<sup>-1</sup>) groups of water extracts of Chijian and Gastrodiae Rhizoma (<italic>P</italic><0.05, <italic>P</italic><0.01), water extract of Gastrodiae Rhizoma with concentration of 400 g·L<sup>-1</sup> and the water extract of Chijian with the concentration of 100 g·L<sup>-1</sup> could promote the proliferation of B lymphocytes induced by lipopolysaccharide. Studies on intestinal microflora showed that compared with the blank group, at the phylum level, the water extracts of Chijian and Gastrodiae Rhizoma increased the relative abundance of Bacteroidetes and decreased the relative abundance of Firmicutes, at the genus level, they increased the relative abundance of <italic>Prevotellaceae</italic>_UCG-001 and <italic>Ruminococcaceae</italic>_UCG-005, and decreased the relative abundance of <italic>Anaerotruncus</italic>, unclassified_<italic>f</italic>_<italic>Erysipelotrichaceae</italic> and<italic> Candidatus</italic>_<italic>Stoquefichus</italic>.<italic> </italic>These intestinal bacteria were related to the immune system, cell proliferation, and metabolism regulation. Conclusion:The total contents of 6 components in the powder, the ethanol and the water extracts of Chijian are higher than or close to those of the corresponding samples of Gastrodiae Rhizoma, the pharmacological activity of Chijian water extract is similar to that of Gastrodiae Rhizoma water extract, indicating that Chijian is worthy of further research and development.

19.
Article in Chinese | WPRIM | ID: wpr-906327

ABSTRACT

Objective:To explore the inhibitory effect and mechanism of Jingulian extract (JGL) on inflammation. Method:The following groups were set up in this study: a control group (10% fetal bovine serum), a lipopolysaccharide (LPS) model group (0.5 mg·L<sup>-1</sup>), and JGL groups (10, 20, 40, 60, 80, 120, 160, 200, 250, 300 mg·L<sup>-1</sup> + 0.5 mg·L<sup>-1 </sup>LPS). The RAW264.7 cells were cultured for 24 hours. Cell proliferation was detected by cell counting kit-8 (CCK-8) assay. Nitric oxide (NO) release was detected by Griess assay. The release of cytokines interleukin (IL)-1<italic>β</italic>, IL-6, IL-10, and tumor necrosis factor (TNF)-<italic>α</italic> was determined by enzyme linked immunosorbent assay (ELISA). The expression of inducible nitric oxide synthase (iNOS) and intraprostaglandin peroxidase synthase 2 (PTGS2)/cyclooxygenase-2 (COX-2) was measured by real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR) and the activation of key proteins in the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway by Western blot. Result:Compared with the control group, LPS (0.5 mg·L<sup>-1</sup>)could promote the proliferation of RAW264.7 cells after stimulation for 24 hours (<italic>P</italic><0.01). Compared with the model group, JGL had no significant effect on cell proliferation. Compared with the control group, LPS (0.5 mg·L<sup>-1</sup>)increased the release of NO, IL-1<italic>β</italic>, IL-6, IL-10, and TNF-<italic>α</italic> (<italic>P</italic><0.01). Compared with the model group, JGL (20-300 mg·L<sup>-1</sup>)inhibited the release of NO in a dose-dependent manner after stimulation for 24 hours (<italic>P</italic><0.05) and reduced IL-1<italic>β</italic>, IL-6, and IL-10 (<italic>P</italic><0.05, <italic>P</italic><0.01), but no obvious inhibition on the release of TNF-<italic>α</italic> was observed. LPS (0.5 mg·L<sup>-1</sup>) could induce the expression of iNOS and PTGS2/COX-2 genes as compared with the control group (<italic>P</italic><0.05, <italic>P</italic><0.01). JGL could down-regulate the mRNA expression of iNOS and PTGS2/COX-2 genes as compared with the model group (<italic>P</italic><0.05, <italic>P</italic><0.01). LPS (0.5 mg·L<sup>-1</sup>) could activate the PI3K/Akt pathway (<italic>P</italic><0.01) as compared with the control group, while JGL (10, 20, 40, and 80 mg·L<sup>-1</sup>) decreased the expression of PI3K-p110, p-p85, and p-Akt (<italic>P</italic><0.01), and inhibited the activation of PI3K/Akt pathway. Conclusion:JGL extract could significantly inhibit the inflammatory response and activation of the PI3K/Akt pathway induced by LPS in RAW264.7 cells. The anti-inflammatory effect was related to the inhibition of the PI3K/Akt pathway.

20.
Article in Chinese | WPRIM | ID: wpr-906284

ABSTRACT

Objective:To explore the effect of chemical compound of aconitum alkaloid on the lipopolysaccharide (LPS)-induced inflammatory response of RAW264.7 macrophages and investigate its mechanism. Method:The chemical compounds of Aconitum Kusnezoffii Reichb were collected from TCMSP database with consideration of oral bioavailability (OB)≥30% and drug-likeness (DL)≥0.18. The potential targets of each chemical component were predicted with use of Pubchem database and Swiss Target Prediction database. Rheumatoid arthritis (RA) targets were collected from GeneCards database and selected by intersection screening. Gene ontology (GO) classification enrichment and Pathway enrichment analysis were carried out with use of DAVID database. Cytoscape was used to construct "Chemical Compound-Potential Targets-Pathway-Disease" network. Protein-protein interaction (PPI) network was constructed by using STRING database and Cytoscape software. RAW264.7 macrophages were stimulated by LPS to establish macrophage inflammation model <italic>in vitro</italic>. Western blot was used to detect the effects of chemical compounds on the expression of tumor necrosis factor-<italic>α</italic> (TNF-<italic>α</italic>) and cyclooxygenase-2 (COX-2) in RAW264.7 cells induced by LPS, as well as on the expression of JAK kinase and nuclear transcription factor- kappa B (NF-<italic>κ</italic>B) signal pathway. Result:A total of 27 chemical compounds were obtained by searching TCMSP database and consulting literature (OB≥30%, DL≥0.18). 12 chemical compounds were obtained after screening. 177 potential targets were obtained after database prediction and screening, and 97 targets were obtained as potential targets for the treatment of RA after intersection between 177 potential targets and 4 329 RA targets. A total of 32 biological processes (BP), 5 cellular components (CC), and 12 molecular functions (MF) were enriched by DAVID database. The construction of network topology map showed that different chemical compounds can act on the same target and the same chemical compound can also act on different targets in the treatment of RA. Aconitum alkaloid can be connected with the same pathway through different targets or with different pathways through the same target, indicating that different targets may have synergistic effect, which fully reflected the complex multi-compound, multi-targets and multi-pathways mechanism. Different concentrations of LPS in stimulation (0-200 μg·L<sup>-1</sup>) can significantly up-regulate the expression of COX-2 protein in RAW264.7 macrophages (<italic>P</italic><0.05), indicating that the inflammatory model was successful. Compared with the normal group, the expression of TNF-<italic>α</italic> and COX-2 protein in the inflammatory model of RAW264.7 cells increased significantly(<italic>P</italic><0.05), while the expression of TNF-<italic>α</italic> and COX-2 protein in bulleyaconitine A(BLA), songorine, yunaconitine and karacoline groups decreased in varying degrees compared with the model group (<italic>P</italic><0.05). Compared with the normal group, the expression of IRAK4, NF-<italic>κ</italic>B, JAK1 and STAT3 in the inflammatory model of RAW264.7 cells were significantly increased (<italic>P</italic><0.05), while such levels in BulleyaconitineA, songorine, yunaconitine and Karacoline groups were significantly lower than those in the model group(<italic>P</italic><0.05). Conclusion:Based on systematic pharmacology and <italic>in vitro</italic> experiments, the related targets and signal pathways were analyzed to provide new insights into the pathogenesis of RA, reveal the molecular mechanism of aconitum alkaloid in the treatment of RA, and provide new ideas for the application of Mongolian medicine in modern medicine.

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