Assessing the Effect of Portulaca oleracea Seed Extract for Alleviating Collagen Type-II Induced Arthritis in Female Wistar Rat

Rheumatoid arthritis is a sign of progressive degradation of cartilage, subchondral bone, and small joints, as well as the persistence of synovitis and the formation of pannus. This research intends to assess the purported anti-arthritic effects of an extract from the seeds of Portulaca oleracea. Female Wistar albino rats (140–200 g) were used and assigned to five groups: Group I administrated NS (10 ml/kg), Group II received 0.2 ml of CoII-IFA, Group III received 300 mg/kg of fish oil, and Groups IV & V administrated 100 and 200 mg/kg of methanolic extract of Portulaca oleracea (MePO). During the experiment, the rats' weight, arthritic score, and footpad oedema were evaluated to determine the severity of their arthritis. Later, blood samples were collected from the animals, which were then analysed for haematological, pro-inflammatory, antioxidant, and histological parameters. A dose-dependent reduction was seen in rats treated with a methanolic extract of Portulaca seeds. Levels of haematological and pro-inflammatory cytokines were considerably reduced by treatment. Although both the standard drug and 200 mg/kg of MePO had anti-inflammatory effects, the latter's were more pronounced at this dose. When looking at the two side by side, results showed that the treatment groups of RBC, WBC, NL-ratio, and ML-ratio levels were normalised. Further histology confirmed the reduction of joint deformity, oedema, formation of pannus, and infiltration of neutrophils in the MePO groups in contrast to arthritic rats. It is hypothesised that Portulaca oleracea may reduce the arthritis and can be used as an adjuvant therapy or incorporate it into your diet with the main course of treatment.

GASTRO-PROTECTIVE EFFECT OF ROOT EXTRACT OF BARLERIA BUXIFOLIA ON ASPIRIN INDUCED GASTRIC ULCER IN RATS

The key assessment of the study was evaluating the gastro-protective properties of Barleria buxifolia root extract (REBB) in ulcerative rats. The roots of Barleria buxifolia have properties like antioxidant, anti-inflammatory, and other health benefits. Induction of gastric ulcers was done with aspirin (150 mg/kg, b.w., p.o) for 3 days and was accompanied by treatment with REBB (200 & 400 mg/kg orally) for 15 days. Ranitidine (20 mg/kg, orally) was received as the standard treatment for 14 days. Ulcer index, percent inhibition of ulceration, lipid peroxidation (LPO), TNF-? levels, and histopathological examination of the gastric mucosa were measured. Aspirin-induced stomach ulcers were seen in 100% of the groups, whereas other animal groups, aside from the control group possessed relatively comparable inductions. Ulcer number, ulcer index (p<0.01), and LPO (p<0.05) shown significant reduction in the 400mg/kg and Ranitidine (20mg/kg) when compared to Aspirin induced control group. Whereas the Ulcer score (p<0.001) and TNF-? (p<0.05) showed significant reduction in Ranitidine treated group but showed objectively improvement but statistically non-significant results in 200mg/kg and 400mg/kg REBB. Hence it was found that Baleria buxifolia root extract effectively mitigates aspirin-induced gastric ulceration in rats, underscoring its potential as a gastro protective agent.

EXPLORING THE THERAPEUTIC POTENTIAL OF AM114: A BORONIC CHALCONE DERIVATIVE INDUCE APOPTOSIS AND SUPPRESS PROINFLAMMATORY CYTOKINES AND CHEMOKINES IN INTERLEUKIN-1? STIMULATED HUMAN THP-1 DERIVED MACROPHAGES

Objective: Chalcones and their derivatives display a wide range of pharmacological activities. This study examined the effects of AM114, a boronic-chalcone derivative, on human THP-1-derived macrophages with and without interleukin-1? (IL-1?) stimulation. Methods: AM114 and Aspirin-treated THP-1-derived macrophages underwent activation with or without interleukin-1?. The IC50 concentrations of AM114 and Aspirin were determined through an MTT test. Apoptosis was measured using various techniques, including staining with acridine orange/Ethidium bromide, Hoechst 33342, and rhodamine 123 assays. Caspase-3 activity was measured using the spectrofluorimetric technique, while DNA fragmentation was assessed via agarose gel electrophoresis. Pro-inflammatory cytokines such as interleukin-6 (IL-6) and chemokines like interleukin-8 (IL-8) were measured using enzyme-linked immunosorbent assays.Results: AM114 and Aspirin showed dose-dependent cytotoxic effects on THP-1 macrophages. Induction of apoptosis was detected in AM114-treated THP-1 macrophages activated with IL-1? compared to macrophages without IL-1?. The gradation of dye uptake, membrane blebbing, increased caspase-3 activity, and DNA fragmentation ensures the induction of apoptosis, which indicates the cell's morphological changes, biochemical processes, and mitochondrial activity. Treating AM114 in IL-1?-activated THP-1 macrophages significantly reduced pro-inflammatory cytokines (IL-6) and chemokines (IL-8), suggesting its anti-cytokine potential in inflammatory diseases.Conclusion: The study results emphasize that AM114 could act as an anti-inflammatory agent by triggering apoptosis and reducing the release of cytokines and chemokines in inflammatory conditions. As a result, it may be used as a therapeutic option for inflammatory diseases.

The Effect on skin papillomas by administration of Fruits of Momordica dioica extract in DMBA/Croton oil induced benign cancer in mice

Skin papillomas are not cancerous, but they can trigger an inflammatory disorder and irritate the area where the wart is present. The study was to find natural agents that prevent and/or cure skin papillomas. The fruits of Momordica dioica have been known to contain potent phytoconstituents that have anti-inflammatory, cardiovascular, diabetic, oncological, and other health benefits. To investigate inhibition of skin papilloma’s by administration of a fruit extract of Momordica dioica in DMBA/Croton oil-induced benign cancer in mice. Induction of papilloma’s by topical application of DMBA/Croton oil for 4-5 weeks; after confirmation of papilloma's, the Momordica dioica and 5-Flu10 mg/kg were administered. Body weight, tumor incidence, cumulative number of tumors, tumor yield, average latency time period, number of papillomas, hematological parameters, antioxidant enzymes, and pro-inflammatory cytokines were measured. 100% tumor incidence was observed in DMBA/croton oil, whereas the other groups except the normal control had quite a similar tumor incidence. The cumulative number, tumor yield, and average latent period have significantly improved with the treatment in skin papilloma mice. Hematological markers and NLR (< 3.0) have improved (p<0.001), which is indicative of reduced acute systemic inflammation. The deviations on LPO and CAT from baselines were corrected by Momordica dioica and found to have a significant impact on their levels (p<0.001). Post-treatment of immunosuppressive markers such as; TGF-beta (p<0.001), TNF-alpha (p<0.001), and IL-6 (p<0.001) were decreased significantly and increased the levels of IFG-g (p<0.001), which correlates with increased immune activation. Hence, the intake of fruits of Momordica dioica inhibits the growth of skin papillomas.

An Investigation of Type II Anti-diabetic activity of Biologically Active phytochemical(s) of Coccinia Indica Fruits in High Sugar Diet Induced Diabetic Rats

Unhealthy diets and lifestyles cause a resistance to and/or relative deficiency of insulin production. Therefore, it is thought worthwhile to develop a natural remedy that may effectively manage the disease symptoms to a certain extent without causing adverse consequences. The objective was to develop the active biological constituent(s) for the use of Coccinia indica and its relationship to treating type II diabetic rats. The soxhlet extraction method was used to get the cocktail of phytochemicals of C. indica by using methanol. The composition of a high-sugar diet, followed by fructose (66%), was used to induce T2DM in rats. The preliminary predictive markers were body weight (pre- and post-treatment), blood glucose level (pre- and post-treatment), serum insulin, and pancreatic insulin. And the secondary outcomes were the pro-inflammatory mediators interleukin 6 (IL-6), transforming growth factor-? (TGF-?), and tumour necrosis factor alpha (TNF alpha). Additionally, pancreatic tissue was used to estimate beta cell mass, size, and necrosis, and the cell supernatant was used to observe superoxide dismutase (SOD), lipid peroxidation (LPO), and catalase (CAT). High sugar diet showed significant increase in body weight (p < 0.01), fasting blood glucose level (p < 0.001), and decrease in serum and pancreatic insulin levels (p < 0.001), whereas rats treated with methanolic extract of C. indica showed significant reduction in post-treatment body weight (p < 0.01), blood glucose levels (p < 0.01), and increase in serum and pancreatic insulin (p < 0.001), especially in higher doses, i.e., 400 mg/kg. Pro-inflammatory cytokines (IL-6, TGF-beta, and TNF-?) can increase insulin resistance, which results in poor glucose homeostasis, which has been reduced by treatment with C. indica (p<0.001). Superoxide radicals and a deficiency in catalase, both of which are linked to diabetes, but the extract of the plant has been shown to enhance the secretion of enzymes SOD and CAT (p<0.001). It has been proven to have a crucial role in the regulation of apoptosis because it lowers oxidative stress and similarly reduces the level of LPO (p<0.01). Additionally, the treated rat pancreas shows islets of Langerhans that are normal in number and size. No necrosis or reduction in size was seen. The current study conclusively shows that the phytoconstituents of C. indica have the potential to tackle long-term health complications and manage symptoms.

Chaihu Shugansan and Its Modified Formulas in Treatment of Functional Dyspepsia： A Review / 中国实验方剂学杂志

Chaihu Shugansan composed of Bupleuri Radix， Paeoniae Radix Alba， Chuanxiong Rhizoma， Aurantii Fructus， Citri Reticulatae Pericarpium， Cyperi Rhizoma， and Glycyrrhizae Radix et Rhizoma has the effects of soothing liver， relieving depression， regulating Qi movement， and relieving pain. It is a classic formula for treating gastric distension recommended by doctors of later ages. This article systematically reviews the clinical application and basic experimental progress of Chaihu Shugansan in the treatment of functional dyspepsia. In modern clinical practice， Chaihu Shugansan and its modified formulas are used to treat functional dyspepsia， and they can be applied in combination with other formulas （Si Junzitang， Jinlingzisan， Zhizhuwan， etc.）， western medicine （domperidone tablets， deanxit， Saccharomyces boulardii， etc.）， traditional Chinese medicine （TCM） acupuncture and other therapies. The results of clinical studies have shown that Chaihu Shugansan and its modified formulas can significantly reduce the Hamilton depression scale （HAMD） score， Hamilton anxiety scale （HAMA） score， and TCM syndrome score， ameliorate the symptoms， improve the quality of life， and decrease the recurrence rate. The experimental pharmacological studies have demonstrated that Chaihu Shugansan can inhibit the autophagy of Cajal interstitial cells， regulate the endoplasmic reticulum stress signaling pathway， and modulate the brain-gut peptide level to improve the gastrointestinal motility. Chaihu Shugansan can inhibit the expression of 5-hydroxytryptamine in the colon tissue and reduce the abdominal withdrawal reflex （AWR） score to improve visceral hypersensitivity. Furthermore， Chaihu Shugansan can lower the levels of pro-inflammatory cytokines such as interleukin-6 and tumor necrosis factor-α to repair duodenal mucosal inflammation. In addition， it can regulate intestinal flora to maintain intestinal flora balance. The main active ingredients such as saikosaponin， paeoniflorin， hesperidin， and naringin in Chaihu Shugansan can exert anti-inflammatory， antioxidant， and antimicrobial effects.

Antitumor and immunomodulatory activity of fucoidan from the brown alga Lessonia trabeculata (Lessoniaceae) on breast cancer spheroids / Actividad antitumoral e inmunomoduladora de fucoidan del alga parda Lessonia trabeculata (Lessoniaceae) en esferoides de cáncer de mama

Abstract Introduction: The therapeutic benefits of the brown algae fucoidan in the treatment of breast cancer have attracted considerable interest in recent years. However, research using spheroids which provide relevant results in trials for antitumor and immunomodulatory products because they adequately simulate the tumor microenvironment, is limited. Objective: To evaluate the antitumor and immunomodulatory activity of Lessonia trabeculata fucoidan (LtF), native to the Peruvian Sea, on two types of multicellular tumor spheroids. Methods: The study was conducted from January to December 2021. Two types of spheroides were elaborated: from 4T1 tumor cells (MTS), and from 4T1 tumor cells+mouse splenocytes (MTSs). The antitumor activity of LtF was evaluated in MTS by quantifying cell viability with MTT. Immunomodulatory activity was determined in MTSs using the IC50 for two types of treatment: simple, fucoidan alone (LtF) and combined, fucoidan+doxorubicin (LtF+Dox). Pro-inflammatory (TNF-α, IL-6) and anti-inflammatory (IL-10, TGF-β) cytokine production was quantified by sandwich ELISA 72 h after treatment. Dox was used as positive control in all assays. Results: LtF exerted antitumor activity as evidenced by increased necrotic zone and cell debris formation compared to the untreated control. Antitumor activity was concentration dependent between 100 and 6 000 μg/ml. In MTSs, simple treatment increased IL-6 and decreased IL-10 and TGF-β production. The combined treatment significantly reduced TGF-β production. In both treatments and Dox, there was an increase in IL-6 compared to the untreated control. The highest production of IL-10 and TGF-β was observed in the untreated control, compatible with a highly immunosuppressive tumor microenvironment. Conclusions: LtF is a good candidate for the treatment of breast cancer and can immunomodulate the tumor microenvironment alone or in combination with Dox.

Resumen Introduccción: Los beneficios terapéuticos del fucoidan de algas pardas en el tratamiento del cáncer de mama han despertado gran interés en los últimos años. Sin embargo, las investigaciones con esferoides son limitadas, éstos proporcionan resultados relevantes en ensayos de productos antitumorales e inmunomoduladores porque simulan adecuadamente el microambiente tumoral. Objetivo: Evaluar la actividad antitumoral e inmunomoduladora del fucoidan de Lessonia trabeculata (LtF), nativa del Mar Peruano, en dos tipos de esferoides tumorales multicelulares. Métodos: El estudio se realizó de enero a diciembre de 2021. Se elaboraron dos tipos de esferoides: con células tumorales 4T1 (MTS) y con células tumorales 4T1+esplenocitos de ratón (MTSs). La actividad antitumoral de LtF se evaluó en MTS cuantificando la viabilidad celular con MTT. La inmunomodulación se determinó en MTSs utilizando la IC50 para dos tipos de tratamiento: simple, fucoidan solo (LtF) y combinado, fucoidan+doxorubicina (LtF+Dox). La producción de citoquinas proinflamatorias (TNF-α, IL-6) y antiinflamatorias (IL-10, TGF-β) se cuantificó mediante ELISA sándwich 72 h post-tratamiento. En todos los ensayos se utilizó Dox como control positivo. Resultados: En los MTS, el LtF ejerció actividad antitumoral evidenciada por aumento de la zona necrótica y formación de restos celulares respecto al control no tratado. La actividad antitumoral fue concentración-dependiente entre 100 y 6 000 μg/ml. En los MTSs, con el tratamiento simple se incrementó IL-6 y disminuyeron IL-10 y TGF-β. El tratamiento combinado redujo significativamente la producción de TGF-β. Los dos tratamientos y Dox incrementaron IL-6 respecto al control no tratado. La mayor producción de IL-10 y TGF-β se observó en los no tratados, compatible con un microambiente tumoral altamente inmunosupresor. Conclusiones: El LtF es un buen candidato para tratar el cáncer de mama y puede inmunomodular el microambiente tumoral solo o en combinación con Dox.

Mecanismos y dianas terapéuticas del inflamasoma NLRP3 en la miocardiopatía diabética / NLRP3 inflammasome mechanism and therapeutic targets in diabetic cardiomyopathy

Resumen La cardiomiopatía diabética es una complicación grave de la diabetes causada por estrés oxidativo, inflamación, resistencia a la insulina, fibrosis miocárdica y lipotoxicidad. Se trata de un padecimiento insidioso, complejo y difícil de tratar. El inflamasoma NLRP3 desencadena la maduración y liberación de citoquinas proinflamatorias, participa en procesos fisiopatológicos como la resistencia a la insulina y la fibrosis miocárdica, además de estar estrechamente relacionado con la aparición y progresión de la cardiomiopatía diabética. El desarrollo de inhibidores dirigidos a aspectos específicos de la inflamación sugiere que el inflamasoma NLRP3 puede utilizarse para tratar la cardiomiopatía diabética. Este artículo pretende resumir el mecanismo y las dianas terapéuticas del inflamasoma NLRP3 en la cardiomiopatía diabética, así como aportar nuevas sugerencias para el tratamiento de esta enfermedad.

Abstract Diabetic cardiomyopathy (DCM) is a serious complication of diabetes caused by oxidative stress, inflammation, insulin resistance, myocardial fibrosis, and lipotoxicity; its nature is insidious, complex and difficult to treat. NLRP3 inflammasome triggers the maturation and release of pro-inflammatory cytokines, participates in pathophysiological processes such as insulin resistance and myocardial fibrosis, in addition to being closely related to the development and progression of diabetic cardiomyopathy. The development of inhibitors targeting specific aspects of inflammation suggests that NLRP3 inflammasome can be used to treat diabetic cardiomyopathy. This paper aims to summarize NLRP3 inflammasome mechanism and therapeutic targets in diabetic cardiomyopathy, and to provide new suggestions for the treatment of this disease.

Anti-inflammatory and antioxidant potential of Artemisia dracunculus L. aqueousextract against acetic acid induced ulcerative colitis in male Wistar rats

Ulcerative colitis (UC) is one of the two types of inflammatory bowel disease (IBD) which is increasing worldover due to modern life style. Patients with UC are prone to develop colorectal cancer. While the disease severity decides the treatment option, researchers look towards herbal medicines with anti-inflammatory properties for minimal or nil side effects. Artemisia dracunculus L., commonly called Tarragon, is a medicinal herb used in traditional Asian medicine mainly in Iran, India, Pakistan and Azerbaijan due to its special compounds. In this study, we tried to elucidate the effects of aqueous extract of tarragon on acetic acid induced ulcerative colitis (UC) in rats. Male Wistar rats were grouped into four groups of eight each viz., control; experimental control (UC was induced via luminal instillation of 4% acetic acid); and UC induced + aqueous tarragon extract (100 mg/kg) or prednisolone (2 mg/kg) orally for ten consecutive days. Tissue specimens were collected after the experimental period for evaluation of caspase-3 and cyclooxygenase-2 (COX-2) expression by immunohistochemistry. Real-time PCR was used to monitor the levels of IL-1, IL-6 and TNF-? in colonic homogenates. Moreover, the levels of myeloperoxidase, nitric oxide and total antioxidant capacity were measured in colonic homogenates. The results showed that both treatment regimens could similarly reduce the severity of disease symptoms. Treatment with aqueous extract of tarragon caused a better improvement (P <0.05) in the levels of myeloperoxidase enzyme, and total antioxidant capacity of colonic homogenates compared to prednisolone. Nevertheless, the levels of the expression of caspase-3, and COX-2 and TNF-? were reduced in UC rats received prednisolone more than UC rats received aqueous extract of tarragon. The was no statistical difference in the levels of nitric oxide, IL-1 and IL-6 between UC rats received tarragon extract or prednisolone. Overall, these findings suggest that the aqueous extract of tarragon is a promising strategy to control ulcerative colitis. Aqueous extract can also be used as an anti-inflammatory and immune system stimulant in conditions where the immune system is damaged.

Chlorogenic acid derived from Moringa oleifera leaf as a potential antiinflammatory agent against cryptosporidiosis in mice

@#Cryptosporidiosis is a serious illness in immunodeficient patients, and there is still no drug that can completely remove the parasite from the host. The present study represents the first report investigating the impact of the active molecule chlorogenic acid (CGA), naturally isolated from Moringa oleifera leaf extract (EMOLE), on immunosuppressed, Cryptosporidium parvum-infected BALB/c mice. Mice were divided into five groups: normal mice, infected immunosuppressed mice, and infected immunosuppressed mice treated with EMOLE, CGA, and nitazoxanide (NTZ) drugs. Parasitological, immunological, and histopathological investigations were recorded besides differences in the mice’ body weight. Infected control mice showed elevated levels of oocyst shedding throughout the study. The EMOLE- and CGA-treated groups showed 84.2% and 91.0% reductions in oocyst shedding, respectively, with no significant difference compared to the drug control. The inflammatory markers IFN-γ, IL-6, IL-1β, and TNF-α were significantly higher in the infected control group. Treatment with 300 mg/kg/day of EMOLE or 30 mg/kg/day of CGA significantly downregulated pro-inflammatory cytokine levels compared to the infected group, although they did not change significantly compared to the NTZ-treated group. Histopathology of intestinal sections showed inflammatory and pathological changes in the infected control group. Low-grade tissue changes and an obvious improvement in villi structure were seen in mice treated with CGA. This study highlighted the role of CGA, isolated and purified from EMOLE, as an effective anti-inflammatory agent in eradicating C. parvum infection.

Relationship between serum MLT and Klotho protein and pro-inflammatory factors and efficacy in patients with primary angle-closure glaucoma / 国际检验医学杂志

Objective To investigate the relationship of serum melatonin(MLT)and Klotho protein with pro-inflammatory factors and the effect of surgical treatment in patients with primary angle-closure glaucoma(PACG).Methods A total of 149 PACG patients admitted to the Zigong Fourth People's Hospital from June 2018 to June 2021 were selected as the case group,and 149 healthy people who underwent physical examina-tion in the hospital during the same period were selected as the control group.The levels of MLT,Klotho pro-tein,interleukin-6(IL-6),interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were compared be-tween the two groups.Pearson correlation analysis was used to analyze the correlation between MLT,Klotho protein and pro-inflammatory factors.Univariate and multivariate Logistic regression models were used to an-alyze the influencing factors of surgical treatment.Results Compared with the control group,the levels of MLT and Klotho protein in the case group were decreased(P<0.05),and the levels of IL-1β,IL-6 and TNF-α were increased(P<0.05).Pearson correlation analysis showed that serum MLT and Klotho protein were negatively correlated with the levels of pro-inflammatory factors IL-1β,IL-6,and TNF-α(P<0.05).After 12 months of surgical treatment,30 patients(20.13%)had no effect of surgical treatment(ineffective group).119 patients(79.87%)had effective surgical treatment(effective group).The univariate analysis showed that compared with the effective group,the preoperative intraocular pressure(IOP),proportion of chronic PACG,preoperative best corrected visual acuity,proportion of preoperative Angle adhesion>160° were significantly increased(P<0.05),and the preoperative anterior chamber depth,MLT,and Klotho protein levels were de-creased in the ineffective group(P<0.05).Multivariate Logistic regression analysis showed that elevated pre-operative IOP,chronic PACG,decreased MLT and Klotho levels were independent risk factors for ineffective surgical treatment(P<0.05).Conclusion serum MLT,Klotho protein levels in patients with PACG,closely associated with proinflammatory factor,is the related factors influencing the effect of surgical treatment.

Astaxanthin protects against hearing impairment in diabetic rats

Abstract Objectives: Diabetes Mellitus (DM) causes an increase in oxidative stress that leads to deterioration in auditory functions. Astaxanthine (AST) is known to have strong antioxidant effects. In this study, the aim is to investigate the effect of AST against hearing loss that is due to DM. Methods: This study is an experimental animal study. The study was designed in four groups with 8 animals (n = 8) in each group. The groups were as follows; Control Group (CNT), Diabetic Group (DM), AST applied diabetic group (DM+AST), and AST applied non-diabetic group (AST). Streptozotocin was applied in rats to induce DM. AST was administered by oral gavage. Auditory Brainstem Responses (ABR) and Distortion Product Otoacoustic Emissions (DPOAE) tests were performed on several days of the study. At the end of the study, pro-inflammatory cytokine levels were analyzed in cochlear tissue samples, and Glutathione Peroxidase (GPx), Superoxide Dismutase (SOD), Catalase (CAT) and Malondialdehyde (MDA) levels were measured. Results: When the findings obtained in the ABR and DPOAE tests in the DM group, it was observed that there was a significant deterioration in the hearing sense. This deterioration was not observed in the DM+AST group. In the DM group, GPx, SOD and CAT levels decreased and MDA levels increased in blood and cochlear tissue. Compared to the DM group, it was noted that antioxidant enzyme levels increased and MDA levels decreased in the DM+AST group. Cochlear tissue pro-inflammatory cytokine levels, which increased with DM, were significantly decreased in the DM+AST group. Conclusion: Even though the effects of AST were investigated in a diabetic experimental animal model, if this molecule is proven to be effective in diabetic humans, it can be considered an adjunct therapeutic option with its antioxidant effects. Level of evidence: The level of evidence of this article is 5. This article is an experimental animal and laboratory study.

Efecto del extracto hidroalcohólico de la planta Lampaya medicinalis Phil . sobre marcadores inflamación inducidos por ácidos grasos en hepatocitos humanos HepG2

RESUMEN Introducción: la enfermedad de hígado graso no alcohólico (EHGNA) se caracteriza por la acumulación de gotas lipídicas (GL) y sobre expresión de la proteína de GL Perilipina 1 (PLIN1) en los hepatocitos. En su patogénesis y progresión participan NF-ĸB, caspasa-1 y citoquinas proinflamatorias como IL-1β. La medicina popular del norte de Chile utiliza la planta Lampaya medicinalis Phil. (Verbenaceae) contra enfermedades. Objetivo: evaluar el efecto de un extracto hidroalcohólico de lampaya (EHL) sobre la expresión de marcadores inflamatorios y proteínas asociadas a las GL en hepatocitos tratados con ácidos grasos. Métodos: se incubó hepatocitos HepG2 con 0,66 mM de ácido oleico (AO) y 0,33 mM de ácido palmítico (AP) por 24 o 48 horas en presencia o no de EHL. Se evaluó la expresión proteica de NF-ĸB, PLIN1 y caspasa-1 por Western blot y la expresión de ARNm de IL-1β por qPCR. Resultados: los hepatocitos tratados por 48 h con AO/AP mostraron un aumento en la expresión de IL-1β que fue revertido por la co-incubación con EHL. Conclusión: estos antecedentes aportan nueva evidencia respecto a la actividad biológica del EHL en un modelo de alteraciones metabólicas e inflamatorias, asociadas a la EHGNA, inducidas por AO/AP en hepatocitos humanos.

ABSTRACT Introduction: Nonalcoholic fatty liver disease (NAFLD) is characterized by the accumulation of lipid droplets (LD) and overexpression of the LD-associated protein Perilipin 1 (PLIN1). NF-ĸB, caspase-1 and proinflammatory cytokine such as IL-1β participate in the pathogenesis and progression of NAFLD. Traditional medicine in northern Chile uses the plant Lampaya medicinalis Phil. (Verbenaceae) against diseases. Objective: To evaluate the effect of a hydroalcoholic extract of lampaya (HEL) on the expression of inflammatory markers and LD-associated proteins in hepatocytes treated with fatty acids. Methods: HepG2 hepatocytes were incubated with 0.66 mM oleic acid (OA) and 0.33 mM palmitic acid (PA) for 24 or 48 h in the presence or not of HEL. The protein expression of NF-ĸB, PLIN1 and caspase-1 was evaluated by Western blot while the mRNA expression of IL-1β was assessed by qPCR. Results: hepatocytes treated for 48 h with OA/AP showed an increase in IL-1β expression that was reversed by co-incubation with HEL. Conclusion: These antecedents provide new evidence regarding the biological activity of HEL in a model of metabolic and inflammatory alterations, associated with NAFLD, induced by OA/PA in human hepatocytes.

Polissacarídeos de Lentinula edodes suprimem a expressão de citocinas pró-inflamatórias e de colite em camundongos / Lentinula edodes polysaccharides suppressed pro-inflammatory cytokines expression and colitis in mice

ABSTRACT Background: Polysaccharides from edible mushrooms possess immunomodulatory, anti-inflammatory, and anti-tumor activities. Recent studies indicated that necroptosis plays a role in the pathogenesis of inflammatory diseases and mediates increased expression of inflammatory cytokines. Objective: Therefore, it is imperative to determine the impact of polysaccharide extract from Lentinula edodes (L. edodes) on inflammatory cytokines in experimental model of colitis in mice. Methods: Female C57BL/6 mice divided into three or four mice per group were used for this study. Polysaccharide sample was orally administered to mice prior to (7 days) and during colitis induction with 2.5% dextran sodium sulfate (7 days), followed by additional 3 days of administration. Changes in body weight and colon length were used as markers for colitis, and pro-inflammatory cytokines and tumor necrosis factor receptor 1 (TNFR1) expressions, as well as necroptosis were analyzed in the colon of colitis mice. Data obtained were analysed by Tukey-Kramer and two-tailed standard t tests. Results: The results indicated that the polysaccharide sample suppressed colitis in mice using effects on the body weight and colon length as markers. Also, it was demonstrated that necrostatin-1, a specific inhibitor of necroptosis, suppressed the expression of interleukin (IL)-8, a pro-inflammatory chemokine, in Caco-2 cells induced necroptosis induced by zVAD and TNF-α, an indication that necroptosis may be involved in the expression of pro-inflammatory cytokines. Moreover, the polysaccharide sample suppressed the expression of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-α, IL-6, IL-1β, and interferon (IFN)-γ in the colon of mice. Conclusion: These results suggested that the suppressive effects of the polysaccharide sample on inflammatory cytokines expression may contribute to its anti-colitis effect, and so may serve as a potent therapeutic agent against inflammatory bowel disease.

RESUMO Contexto: Polissacarídeos de cogumelos comestíveis possuem atividades imunomodulatórias, anti-inflamatórias e anti-tumorais. Estudos recentes indicaram que a necroptose desempenha um papel na patogênese de doenças inflamatórias e regula o aumento da expressão de citocinas inflamatórias. Objetivo: Torna-se imprescindível determinar o impacto do extrato de polissacarídeo de Lentinula edodes (L. edodes) em citocinas inflamatórias em modelo experimental de colite em camundongos. Métodos: Foram utilizados para este estudo os camundongos C57BL/6 femininos divididos em três ou quatro camundongos por grupo. A amostra de polissacarídeo foi administrada oralmente em camundongos antes (7 dias) e durante a indução de colite com sulfato de dextran sulfato de sódio (7 dias), seguido por 3 dias adicionais de administração. Alterações no peso corporal e comprimento do cólon foram utilizadas como marcadores para colite, e citocinas pró-inflamatórias e tumores receptor fator 1 (TNFR1), bem como necroptose foram analisadas no cólon de camundongos colite. Os dados obtidos foram analisados por testes Tukey-Kramer e testes padrão t de duas caudas. Resultados: Os resultados indicaram que a amostra de polissacarídeo suprimiu colite em camundongos usando efeitos sobre o peso corporal e o comprimento do cólon como marcadores. Além disso, foi demonstrado que a necrostatina-1, inibidora específica da necroptose, suprimiu a expres são de interleucina (IL)-8, uma quimiocina pró-inflamatória, em células caco-2 induzidas necroptose induzidas por zVAD e TNF-α, uma indicação de que a necroptose pode estar envolvida na expressão de citocinas pro-inflamatórias. Além disso, a amostra de polissacarídeo suprimiu a expressão de citocinas pró-inflamatórias, como o fator de necrose tumoral (TNF)-α, IL-6, IL-1β e interferon (IFN)-γ no cólon dos camundongos. Conclusão: Esses resultados sugeriram que os efeitos supressivos da amostra de polissacarídeo na expressão de citocinas inflamatórias podem contribuir para o seu efeito anti-colite, podendo, portanto, servir como um potente agente terapêutico contra doença inflamatória intestinal.

Taxifolin attenuates inflammation via suppressing MAPK signal pathway in vitro and in silico analysis / 中草药·英文版

Objective: Taxifolin is a natural flavonoid compound that can be isolated from onions, grapes, oranges and grapefruit. It also acts as a medicine food homology with extraordinary antioxidant and anti-inflammatory activity. This study aims to explain the protective effects and potential mechanisms of taxifolin against inflammatory reaction. Methods: Levels of interleukin (IL)-6, IL-1β and intracellular reactive oxygen species (ROS) were assessed in different time after the treatment of taxifolin in RAW264.7 cells induced by lipopolysaccharide (LPS). Subsequently, the mRNA and protein levels of inducible nitric oxide synthase (iNOS), vascular endothelial growth factor (VEGF), cyclooxygenase (COX)-2, tumor necrosis factor (TNF)-α and the phosphorylation expression levels of the MAPK signal pathway were also evaluated. A silico analysis was used to explain the binding situation for the investigation of taxifolin and MAPK signal pathway. And then MAPK inhibitors were used to reveal the expression level of iNOS, VEGF, COX-2 and TNF-α in RAW264.7 cells. Results: It was demonstrated that cell inflammatory damage induced by LPS was significantly alleviated after the treatment of taxifolin. Then, the mRNA and protein levels of iNOS, VEGF, COX-2 and TNF-α were reduced and the phosphorylation expression levels of the MAPK signal pathway were down-regulated remarkably as well. In silico analysis, taxifolin could form a relatively stable combination with MAPK signal pathway. MAPK inhibitors showed increasing or decreasing effect in the mRNA levels of iNOS, VEGF, COX-2 and TNF-α, which suggesting that taxifolin down-regulated iNOS, VEGF, COX-2 and TNF-α expressions were not entirely through the MAPK pathway. Conclusion: This finding demonstrated that taxifolin improved the inflammatory responses that partly involved in the phosphorylation expression level of MAPK signal pathway in RAW264.7 cells exposed to acute stress.

Study on Effect of Notopterol on Dextran Sulfate Sodium-induced Colitis in Mice / 胃肠病学

Background; Immunological dysfunction plays a key role in the pathogenesis of inflammatory bowel disease (I B D). Notopterol is the main ingredient of the traditional Chinese herb medicine Notopterygium ineisum Ting ex H. T. Chang and has anti-inflammatory effect. Aims; To explore the effect of notopterol on dextran sulfate sodium (DSS)-induced colitis in mice. Methods; C57BL/6 mice were randomly divided into normal group, negative control group, model group and notopterol group. Mice in model group and notopterol group were treated with 2% DSS to induce colitis. Mice in negative control group and notopterol group were injected intraperitoneally with notopterol 40 mg/kg, and mice in normal group and model group were injected intraperitoneally with 0. 9% NaCl solution. Mice were sacrificed 10 days later, and disease activity index (DAI) score, colon length and histopathologieal score were determined. The levels of TNF-α, IL-1β, IL-6, IL-17A were assessed by ELISA. The mRNA expressions of IL-17A, RORγt were detected by quantitative PCR. Results; Compared with the normal group, DAI score, histopathologieal score, levels of TNF-α, IL-1β, IL-6, IL-17A, mRNA expressions of IL-17A and RORγt in model group were significantly increased (P < 0. 0 1), and colon length were significantly shortened (P < 0. 0 1). Compared with the model group, notopterol significantly reduced DAI score and histopathologieal score (P<0.01), downregulated levels of TNF-α, IL-1β, IL-6, IL-17A (P<0.01), inhibited the mRNA expressions of IL-17A, ROR-γt (P < 0. 0 1), and greatly recovered colon length (P < 0. 0 1). Conclusions; Notopterol has protective effects on DSS-induced colitis in mice. The mechanism may be related to reducing the secretion of pro-inflammatory cytokines and inhibiting the function and differentiation of Thl7 cells.

Ovarian hormones influence immune response to Staphylococcus aureus infection

ABSTRACT Objective: Staphylococcus aureus infections remain associated with considerable morbidity and mortality in both hospitals and the community. There is little information regarding the role of ovarian hormones in infections caused by S. aureus. The aim of this study was to evaluate the effects of ovariectomy in the immune response induced by S. aureus. Methods: Female mice BALB/c were ovariectomized (OVX) to significantly reduce the level of ovarian hormones. We also used sham-operated animals. The mice were inoculated intraperitoneally with S. aureus. Blood samples were collected for leukocyte count and bacterial quantification. The uterus and spleen were removed and weighed to calculate the uterine and splenic indexes. Lungs were removed and fractionated for immunohistochemical analysis for macrophage detection (anti-CD68) and relative gene expression of IL-6, IL-1β and TNF-α by RT-PCR. Results: Ovariectomy enlarged spleen size and generally increased circulating lymphocytes. OVX females experienced a continuation of the initial reduction of lymphocytes and a monocyte and neutrophil late response compared to shams (p ≥ 0.05). Moreover, OVX females showed neutropenia after 168 h of infection (p ≥ 0.05). Macrophage response in the lungs were less pronounced in OVX females in the initial hours of infection (p ≥ 0.01). OVX females showed a higher relative gene expression of IL-1β, IL-6 and TNF-α in the lung at the beginning of the infection compared to sham females (p ≥ 0.01). Among the uninfected females, the OVX control females showed a higher expression of IL-6 in the lung compared to the sham control females (p ≥ 0.05). In this model, the lack of ovarian hormones caused a minor increase in circulating leukocytes during the initial stage of infection by S. aureus and increased pulmonary gene expression of IL-1β, IL-6, and TNF-α. Ovariectomy alone enlarged the spleen and increased circulating lymphocytes. Ovarian hormones acted as immunoprotectors against S. aureus infection.

Effect of coinfection with influenza virus and bacteria on host damage / Efecto de la coinfección por virus de la influenza y bacterias en el daño al hospedero

Abstract Background: Influenza virus infection is often complicated by a bacterial infection, with this coinfection causing severe pneumonia. If not timely treated, the disease can cause death. Objective: To demonstrate, in animal models, that coinfection with influenza virus and bacteria that affect the respiratory tract causes multisystemic damage. Method: Six groups of mice were formed: a control group, one infected with the influenza virus, two infected with bacteria: Haemophilus influenzae and Streptococcus pneumoniae, respectively; and two co-infected with influenza virus and Haemophilus influenzae or Streptococcus pneumoniae, respectively. Results: Of the six groups of mice, only the group co-infected with influenza virus and Streptococcus pneumoniae showed damage to thoracic and abdominal organs. A decrease in serum cytokine levels was found in all study groups, which was more pronounced in the co-infected mice. Conclusions: The groups of mice infected with Streptococcus pneumoniae or influenza virus alone showed no damage, which indicates that coexistence of these infections caused the damage in the group of co-infected mice.

Resumen Antecedentes: La infección por el virus de la influenza con frecuencia se complica con una infección bacteriana, coinfección que provoca cuadros graves de neumonía, la cual puede ocasionar la muerte si no es tratada en forma oportuna. Objetivo: Demostrar en modelos animales que la coinfección por el virus de la influenza y bacterias que afectan el tracto respiratorio ocasiona daño multisistémico. Método: Se formaron seis grupos de ratones: un grupo control, uno infectado de virus de la influenza, dos infectados de bacterias: Haemophilus influenzae y Streptococcus pneumoniae, respectivamente; y dos coinfectados de virus de la influenza y Haemophilus influenzae y Streptococcus pneumoniae, respectivamente. Resultados: De los seis grupos de ratones, solo en el grupo coinfectado de virus de la influenza y Streptococcus pneumoniae se observó daño en órganos torácicos y abdominales. En todos los grupos se encontró disminución de los niveles séricos de las citocinas, mayor en los ratones coinfectados. Conclusiones: Los grupos de ratones infectados solo de Streptococcus pneumoniae o el virus de la influenza no presentaron daños, lo cual indica que la coexistencia de estas infecciones fue la que ocasionó el daño en el grupo de ratones coinfectados.

Cholinergic anti-inflammatory pathway plays negative regulatory role in early inflammatory and immune responses in septic rats / 南方医科大学学报

OBJECTIVE@#To investigate the role of cholinergic anti-inflammatory pathway (CAP) in neuro-regulation of inflammatory and immune response in the early stage of sepsis.@*METHODS@#Sixty-four SD rats were randomly divided into control Group (=8) with normal feeding without any treatment; sham operation group (=8) with laparotomy but without cecal ligation and puncture (CLP), followed by intraperitoneal injection 50 mg/kg piperacillin 3 times a day for 3 consecutive days; and sepsis group (=48) with CLP-induced sepsis. The rat models of sepsis were randomized into model groups (=16) with intraperitoneal injection of piperacillin (50 mg/kg) and normal saline (1 mL/100 g) for 3 times a day for 3 days; GTS-21 group (=16) with additional intraperitoneal injection of 4 mg/kg GTS-21 (once a day for 3 days); and methyllycaconitine (MLA) group (=16) with intraperitoneal injection of MLA (4.8 mg/kg) in addition to piperacillin (once a day for 3 days). Murine Sepsis Score (MSS) of the rats and short-range HRV analysis were recorded. Three days later, the rats were sacrificed and serum levels of TNF-α, IL-1α, IL-10, IL-6, HMGB1, and sCD14 were measured with ELISA. The percentages of CD4CD25 Treg and TH17 lymphocytes and their ratios were measured using flow cytometry.@*RESULTS@#Compared with the control rats, the septic rats had significantly increased MSS scores and lowered HRV indexes (SDNN, RMSSD, HF, SD1, and SD2; < 0.05); treatment with GTS-21 significantly decreased while MLA increased MSS scores ( < 0.05), but neither of them obviously affected HRV of the rats. Serum levels TNF-α, IL-1α, IL-10, IL-6, HMGB1, and sCD14 and the percentages of CD4CD25 Treg and TH17-positive lymphocytes were significantly higher and Treg/TH17 ratio was significantly lower in the septic rats compared with those in the control group ( < 0.05); treatment with GTS-21 significantly decreased the levels of serum levels of TNF-α, IL-1α, IL-6, HMGB1, and sCD14 and TH17 lymphocyte percentage ( < 0.05), whereas MLA treatment significantly increased serum levels of TNF-α, IL-1α, IL-10, IL-6, HMGB1, and sCD14 and the percentages of CD4 CD25 Treg and TH17-positive lymphocytes and decreased Treg/TH17 ratio in the septic rats ( < 0.05).@*CONCLUSIONS@#CAP plays negative regulatory role in early inflammatory and immune response to sepsis, and some of the HRV indicators can well reflect the regulatory effect of CAP on inflammation and immunity in the septic rats.

Effects of leptin-modified human placenta-derived mesenchymal stem cells on angiogenic potential and peripheral inflammation of human umbilical vein endothelial cells (HUVECs) after X-ray radiation / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Combined radiation-wound injury (CRWI) is characterized by blood vessel damage and pro-inflammatory cytokine deficiency. Studies have identified that the direct application of leptin plays a significant role in angiogenesis and inflammation. We established a sustained and stable leptin expression system to study the mechanism. A lentivirus method was employed to explore the angiogenic potential and peripheral inflammation of irradiated human umbilical vein endothelial cells (HUVECs). Leptin was transfected into human placenta-derived mesenchymal stem cells (HPMSCs) with lentiviral vectors. HUVECs were irradiated by X-ray at a single dose of 20 Gy. Transwell migration assay was performed to assess the migration of irradiated HUVECs. Based on the Transwell systems, co-culture systems of HPMSCs and irradiated HUVECs were established. Cell proliferation was measured by cell counting kit-8 (CCK-8) assay. The secretion of pro-inflammatory cytokines (human granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin (IL)-1α, IL-6, and IL-8) was detected by enzyme-linked immunosorbent assay (ELISA). The expression of pro-angiogenic factors (vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF)) mRNA was detected by real-time quantitative polymerase chain reaction (RT-qPCR) assay. Relevant molecules of the nuclear factor-KB (NF-kB) and Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathways were detected by western blot assay. Results showed that leptin-modified HPMSCs (HPMSCs/leptin) exhibited better cell proliferation, migration, and angiogenic potential (expressed more VEGF and bFGF). In both the single HPMSCs/leptin and the co-culture systems of HPMSCs/leptin and irradiated HUVECs, the increased secretion of pro-inflammatory cytokines (human GM-CSF, IL-1α, and IL-6) was associated with the interaction of the NF-KB and JAK/STAT signaling pathways. We conclude that HPMSCs/leptin could promote angiogenic potential and peripheral inflammation of HUVECs after X-ray radiation.