Quercetin and resveratrol are associated with the downregulation of TNFalpha/NF-kB/inos axis-mediated acute liver injury in rats induced by paracetamol poisoning / La quercetina y el resveratrol están asociados con la regulación decreciente de la lesión hepática aguda mediada por el eje TNF-alfa/NF-kB/inos en ratas inducida por envenenamiento con paracetamol

SUMMARY: Paracetamol (known as acetaminophen, or APAP) poisoning causes acute liver damage that can lead to organ failure and death. We sought to determine that APAP overdose can augment tumor necrosis factor-alpha (TNF-α)/ nuclear factor kappa B (NF-kB)/induced nitic oxide synthase (iNOS) axis-mediated hepatotoxicity in rats, and the anti-inflammatory polyphenolic compounds, quercetin (QUR) plus resveratrol (RES) can ameliorate these parameters. Therefore, we induced acute hepatotoxicity in rats using APAP overdose (2 g/kg, orally) and the protective group of rats were treated with 50 mg/kg QUR plus 30 mg/kg RES for one week before APAP ingestion. Animals were killed at day 8. APAP poisoning caused the induction of hepatic tissue levels of TNF-α, NF-kB, and iNOS, which were significantly (p<0.05) decreased by QUR+RES. QUR+RES, also inhibited liver injury biomarkers, alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Additionally, a link between liver injury and TNF-α /NF-kB / iNOS axis mediated hepatotoxicity was observed. Thus, the presented data backing the conclusion that intoxication by paracetamol increases TNF-α / NF-kB / iNOS axis -mediated hepatotoxicity, and is protected by a combination of quercetin and resveratrol.

El envenenamiento por paracetamol (conocido como acetaminofeno o APAP) causa daño hepático agudo que puede provocar una insuficiencia orgánica y la muerte. El objetivo de este trabajo fue determinar si la sobredosis de APAP puede aumentar la hepatotoxicidad mediada por el eje del factor de necrosis tumoral alfa (TNF-α)/factor nuclear kappa B (NF-kB)/óxido nítico sintasa inducida (iNOS) en ratas, y si el polifenólico antiinflamatorio compuesto por quercetina (QUR) más resveratrol (RES) pueden mejorar estos parámetros. Por lo tanto, inducimos hepatotoxicidad aguda en ratas usando una sobredosis de APAP (2 g/kg, por vía oral). El grupo protector de ratas se trató con 50 mg/ kg de QUR más 30 mg/kg de RES durante una semana antes de la ingestión de APAP. Los animales se sacrificaron el día 8. El envenenamiento con APAP en el tejido hepático provocó la inducción de niveles de TNF-α, NF-kB e iNOS, que se redujeron significativamente (p<0,05) con QUR+RES. QUR+RES, también inhibió los biomarcadores de daño hepático, la alanina aminotransferasa (ALT) y el aspartato aminotransferasa (AST). Además, se observó una relación entre la lesión hepática y la hepatotoxicidad mediada por el eje TNF-α /NF-kB/iNOS. Por lo tanto, los datos presentados respaldan la conclusión de que la intoxicación por paracetamol aumenta la hepatotoxicidad mediada por el eje TNF-α /NF-kB / iNOS, y está protegida por una combinación de quercetina y resveratrol.

Mechanism of Flavonoids from Mulberry Leaves on Insulin Resistance and Inflammatory Response in Diabetic Mice / 中国实验方剂学杂志

ObjectiveTo observe the glucose-lowering， insulin resistance-improving， and anti-inflammatory effects of flavonoids from mulberry leaves （FML） and explore their underlying mechanism. MethodMale db/db mice aged 6-7 weeks were randomly divided into a model group， a high-dose FML group （1.00 g·kg·d-1）， and a low-dose FML group （0.50 g·kg-1·d-1）. C57BL mice of the same age were assigned to the normal group. After six weeks of intervention， fasting blood glucose （FBG）， serum fasting insulin levels （Fins）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， free fatty acid （FFA）， blood creatinine （SCr）， blood urea nitrogen （BUN）， and aspartate aminotransferase （AST） levels were measured， and the homeostasis model assessment of insulin resistance （HOMA-IR） was calculated. Superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px）， and catalase activities in the liver were measured. Morphological changes in the liver were assessed by hematoxylin-eosin （HE） staining. The protein expression of cyclooxygenase-2 （COX-2）， inducible nitric oxide synthase （iNOS）， and nuclear factor-κB （NF-κB） in the liver was detected by Western blot. ResultCompared with the model group， the high-dose and low-dose FML groups showed significant reductions in FBG， Fins， HOMA-IR， IL-6， TNF-α， and FFA levels （P<0.05， P<0.01）， and increased levels of SOD， GSH-Px， and catalase in the liver （P<0.05， P<0.01）. HE staining of the liver in the FML groups showed improved arrangement of hepatocytes， reduced inflammatory cell infiltration， and alleviated cellular steatosis compared with the model group. The protein expression of COX-2， iNOS， and NF-κB in the liver significantly decreased in the FML groups as compared with that in the model group （P<0.05， P<0.01）. ConclusionFML have glucose-lowering and insulin resistance-improving effect， which may be attributed to their regulation of the NF-κB pathway in the liver of diabetic mice， leading to the suppression of the release of COX-2， iNOS， and inflammatory cytokines， thereby improving the inflammatory state.

Suppression of nitrosative stress and inflammation of the knee joint synovium in collagen type ii-induced rheumatoid arthritis by the inhibition of glycogen synthase kinase-3ß / Supresión del estrés nitrosativo e inflamación de la sinovial de la articulación de la rodilla en la artritis reumatoide inducida por colágeno tipo ii mediante la inhibición de la glucógeno sintasa quinasa-3 ß

SUMMARY: Rheumatoid arthritis (RA), an inflammatory autoimmune disease that causes cartilage degradation and tissue destruction, can affect synovial joints such as the knee joint. The link between the nitrosative stress enzyme inducible nitric oxide synthase (iNOS) and the cytokine interleukin-1 (IL-1β) in RA-induced knee joint synovial membrane damage with and without the incorporation of the GSK3β inhibitor TDZD-8 has never been studied. As a result, we used active immunization method with collagen type II (COII) for twenty one days to induce RA in rats. TDZD-8 (1 mg/kg; i.p.) was given daily into matched immunized rats for three weeks after day 21 (COII+TDZD-8). Blood and tissue samples were taken 42 days after immunization. A dramatic increase in rheumatoid factor (RF) blood levels, as well as considerable synovial tissue damage and inflammatory cell infiltration of the synovial membrane, were used to validate the onset of RA following COII immunization. COII immunization increased tissue levels of iNOS protein and IL- 1β mRNA and protein expression, which TDZD-8 suppressed considerably (p<0.0001). Furthermore, there was a significantly (p<0.001) positive correlation between iNOS, inflammatory biomarkers, and RF. We concluded that TDZD-8 reduced RA-induced IL-1β -iNOS axis-mediated arthritis in the rat knee joint synovium.

RESUMEN: La artritis reumatoide (AR), es una enfermedad autoinmune inflamatoria que causa la degradación del cartílago y la destrucción del tejido, pudiendo afectar las articulaciones sinoviales, como la articulación de la rodilla. No se ha estudiado el vínculo entre la óxido nítrico sintasa inducible por la enzima del estrés nitrosativo (iNOS) y la citocina interleucina-1 (IL-1β) en el daño de la membrana sinovial de la articulación de la rodilla provocado por AR con y sin la incorporación del inhibidor de GSK3β TDZD-8. Utilizamos el método de inmunización activa con colágeno tipo II (COII) durante veintiún días para inducir AR en ratas. Se administró TDZD-8 (1 mg/kg; i.p.) diariamente a ratas inmunizadas emparejadas durante tres semanas después del día 21 (COII+TDZD- 8). Se tomaron muestras de sangre y tejido 42 días después de la inmunización. Se observó un gran aumento de los niveles sanguíneos del factor reumatoideo (FR), así como un daño considerable del tejido sinovial e infiltración de células inflamatorias en la membrana sinovial, para validar la aparición de la AR después de la inmunización con COII. La inmunización con COII aumentó los niveles tisulares de la proteína iNOS y la expresión de proteína y ARNm de IL-1β, que TDZD-8 suprimió considerablemente (p<0,0001). Además, hubo una correlación positiva significativa (p<0,001) entre iNOS, biomarcadores inflamatorios y FR. Concluimos que TDZD- 8 redujo la artritis mediada por el eje IL-1β-iNOS inducida por la AR en la sinovial de la articulación de la rodilla de rata.

Mechanism of Total Saponins from Dioscoreae Nipponicae Rhizoma to Treat Gouty Arthritis by Regulating M1/M2 Polarization of Macrophages / 中国实验方剂学杂志

Objective:To illustrate the effect of M1/M2 polarization of macrophages on gouty arthritis models induced with monosodium urate and reveal the molecular mechanism of total saponins from Dioscoreae Nipponicae Rhizoma to treat gouty arthritis. Method:A total of 72 male SD rats were randomly divided into four groups： normal group， model group， total saponin group （160 mg·kg^-1）， celecoxib group （43.3 mg·kg^-1）， with 18 rats in each group. Gouty arthritis models were induced by injecting monosodium urate into ankle joints bilaterally. Histopathology changes of ankle joints were observed by hematoxylin-eosin（HE） staining. Immunohistochemistry method was used to detect the protein expression change of CD68， interleukin-4（IL-4）， inducible nitric oxide synthase （iNOS） and transforming growth factor-<italic>β</italic>₁（TGF-<italic>β</italic>₁）. Result:HE staining results showed that the inflammation of the model group was most obvious on the third day after modeling， and the disease was in the acute stage. On day 5， the inflammation was alleviated， and on day 8， the inflammation was still present but close to normal. The total saponin group and celecoxib group could improve the pathological changes of synovial tissue， and the effect of total saponin group was more obvious. Immunohistochemical results were as follows. Compared with the normal group. The expression of CD68 and iNOS in the model group increased on the 3rd，5th and 8th day of administration （<italic>P</italic><0.01）. Compared with the model group， the total saponins group could reduce the expression of CD68 and iNOS （<italic>P</italic><0.05，<italic>P</italic><0.01）on the 3rd day of administration， and significantly reduced them expression on the 5th and 8th days （<italic>P</italic><0.01）. Compared with the normal group， IL-4 and TGF-<italic>β</italic>₁ expression were increased in the model group when the drug was given for three days（<italic>P</italic><0.01）. Total saponin group could enhance IL-4 expression（<italic>P</italic><0.05）and decreased the TGF-<italic>β</italic>₁ expression（<italic>P</italic><0.01）. Compared with normal group， the expression of IL-4 in the model group decreased on the 5th and 8th day of administration （<italic>P</italic><0.01）， and the expression of TGF-<italic>β</italic>₁ in the model group decreased on the 5th day of administration（<italic>P</italic><0.01）. Compared with the model group， the total saponins group could increase the expression of IL-4 and TGF-<italic>β</italic>₁ at 5 d and 8 d after administration （<italic>P</italic><0.01）. Conclusion:Total saponins from Dioscoreae Nipponicae Rhizoma has the potential effect to treat gouty arthritis by regulating M1/M2 polarization.

Experimental study on the role of IL-10 in improving donor lung function after ex vivo lung perfusion in rats of cardiac death / 器官移植

Objective To evaluate the effect of interleukin (IL)-10 on donor lung function after ex vivo lung perfusion (EVLP) in rats of cardiac death. Methods Twenty adult male SD rats were randomly divided into the simple perfusion group (group A, n=10) and modified perfusion group (group B, n=10). Perfusate A (without IL-10) and perfusate B (supplemented with IL-10) was administered in group A and B, respectively. The EVLP rat models of cardiac death were established. The appearance of donor lung, dry-to-wet (D/W) ratio of donor lung tissues, the function and metabolism of donor lung, the morphology of donor lung and the levels of inflammatory markers of donor lung were statistically compared between two groups. Results After perfusion, evident edema of the whole donor lung, poor compliance and a large amount of edema fluid discharged from the airway were observed in group A, whereas no obvious edema and good compliance were found in group B. Compared with group A, the D/W ratio of lung tissues in group B was higher (P < 0.05). In both groups, the pulmonary vein partial pressure of oxygen reached the peak at 2 h after perfusion, which did not significantly differ between two groups (P > 0.05). In group B, the pulmonary artery pressure was increased at a lower speed and significantly lower after perfusion, and the lactic acid level in the perfusate was significantly lower than those in group A (all P < 0.05). In group A, the alveolar structure was largely destroyed and the cells was rare. In group B, the alveolar structure was relatively normal without evident cell edema. The incidence of cell apoptosis of donor lung was high in group A, whereas no obvious cell apoptosis of donor lung was noted in group B. After perfusion for 4 h, the levels of monocyte chemoattractant protein (MCP)-1 and IL-6 were significantly increased, the IL-4 levels were remarkably decreased (all P < 0.05), but the levels of tumor necrosis factor (TNF)-α, IL-1α and inducible nitric oxide synthase (iNOS) did not significantly change in both groups (all P > 0.05). Conclusions IL-10 may improve the function of donor lung after EVLP in rat of cardiac death by reducing cell apoptosis.

Effect of Dendrobium officinale on Yin deficiency constipation induced by warm-drying medicine in mice / 中国中药杂志

Dendrobium officinale is a sacred product for nourishing Yin and has a clear "thick gastrointestinal" effect. Modern pharmacological studies had found that it could improve gastrointestinal function. This study observed the improvement effect of D. officinale on constipation model mice with Yin deficiency caused by warm-drying medicine. It provided experimental basis for the treatment of Yin deficiency constipation. The male and female ICR mice were randomly divided into normal group, model group, D. officinale high, medium and low dose groups(0.6, 0.4, 0.2 g·kg~(-1)), and phenolphthalein tablets group. The model mice of Yin deficiency constipation were established by gavage with warm-drying medicine. The overall state and body temperature of the mice were observed and recorded. The number of feces, feces weight, fecal moisture content and intestinal propulsion were measured. The morphological damage of colon tissue was observed by hematoxylin-eosin(HE) staining. The expression of inducible nitric oxide synthase(iNOS) in the colon was detected by Western blot and immunohistochemical method. The expression of iNOS mRNA in the colon was detected by Real-time fluorescence quantitative PCR, and the serum cyclic guanosine phosphate(cGMP) level was detected the enzyme-linked immunosorbent assay(ELISA). The results showed that D. candidum could reduce the body temperature of mice with Yin deficiency constipation, increase the number of feces, wet feces, dry feces and intestinal propulsion ability, reduce the expression of iNOS protein and mRNA in the colon, and reduce the content of cGMP in the serum. It showed that D. candidum could improve the symptoms of Yin deficiency constipation mice caused by warm-drying medicine, and the mechanism may be related to reducing the expression of iNOS in the colon and increasing intestinal motility.

Changes and significance of monocytic myeloid-derived suppressor cells during acute phase of Kawasaki disease / 中华微生物学和免疫学杂志

Objective:To investigate the changes of monocytic myeloid-derived suppressor cells (M-MDSC) in children with acute Kawasaki disease (KD) and its roles in the immunological pathogenesis of KD.Methods:A total of 38 children with acute KD were enrolled in the present study and 32 age-matched healthy children were selected as control group. The proportions of HLA-DR -CD11b + CD33 + CD15 -CD14 + M-MDSC and CD4 + CD25 + CD127 - regulatory T cells (Treg) in peripheral blood, concentrations of reactive oxygen species (ROS) and expression of arginase-1 (Arg-1), CD39, CD73, CD40, CD40L and CCR5 at protein levels were detected by flow cytometry. Quantitative real-time PCR was used to evaluate the transcription levels of inducible nitric oxide synthase (iNOS) in M-MDSC and the transcription levels of cytotoxic T-lymphocyte associated antigen 4 (CTLA4) and lymphocyte-activation gene 3 (LAG3) in Treg. Concentrations of NO, CCL3, CCL4, CCL5, IL-10 and TGF-β in the supernatants of cell culture were measured by ELISA. Results:(1) The proportion of HLA-DR -CD11b + CD33 + CD15 -CD14 + M-MDSC, the concentration of intracellular ROS and the expression of iNOS, CD39 and CD73 in M-MDSC decreased significantly in patients with acute KD as compared with those in the control group ( P<0.05), and the concentrations of NO, IL-10 and TGF-β in culture supernatant of M-MDSC were lower than those in the control group upon lipopolysaccharide (LPS) stimulation for 48 h ( P<0.05). All of the aforementioned indexes restored to some extent after intravenous immunoglobulin (IVIG) therapy ( P<0.05). No statistical differences were found in Arg-1 expression between healthy controls and patients with KD before or after IVIG therapy ( P<0.05). (2) CD40 expression on M-MDSC was significantly lower in the acute KD group than in the control group ( P<0.05). The concentrations of CCL3, CCL4 and CCL5 in the culture supernatants of M-MDSC were lower in the acute KD group than in the control group after LPS stimulation ( P<0.05). With IVIG treatment, all of the indexes were up-regulated significantly ( P<0.05), although CD40 expression was still lower in the acute KD group than in the control group ( P<0.05). (3) The proportion of CD4 + CD25 + CD127 -Treg and the expression of CTLA4, LAG3, CD40L and CCR5 reduced significantly in patients with acute KD as compared those in healthy controls ( P<0.05), and all increased remarkably after IVIG therapy ( P<0.05). Pearson correlation analysis showed a positive correlation between the proportions of M-MDSC and Treg in patients with acute KD ( r=0.58, P<0.05). Conclusions:Insufficiency and impaired function of M-MDSC might be a major cause of immune dysfunction in patients with acute KD.

Bioassay guided fractionation of Cyclea peltata using in vitro RAW 264.7 cell culture, antioxidant assays and isolation of bioactive compound tetrandrine

Background: Cyclea peltata is one of the herbs mentioned in ancient scriptures of Ayurveda and is used indifferent types of Ayurvedic gritham preparations. Moreover, in traditional/tribal medicine C. peltata isused as digestive, anti-inflammatory, diuretic and to treat jaundice, digestive disorders, etc.Objective: Activity guided fractionation of C. peltata and in correlation with the levels of bioactivecompound tetrandrine.Materials and methods: Preliminary phytochemical screening, estimation of total alkaloid content,preparation of different extracts of C. peltata (crude extract CP, hexane extract HCP, chloroform extractCCP, methanol extract MCP, alkaloid fraction ACP). In vitro anti-inflammatory studies using RAW264.7 cells and in vitro antioxidant assays of the different extracts of C. peltata. HPTLC estimation oftetrandrine (TET) was carried out using solvent system toluene: ethyl acetate: diethylamine (7.2: 2: 0.8)and isolation of TET from ACP.Results: Preliminary phytochemical studies of C. peltata showed the presence of alkaloid content in allextracts. Whereas, saponins, steroids and terpenoids were detected in CP and CCP. ACP and TET showedsignificant in vitro anti-inflammatory and antioxidant activity when compared to other extracts. ACP andTET (100 mg/ml) treatment significantly inhibited the mRNA expression of iNOS, COX-2, TNF-a in LPStreated RAW 264.7 cells. HPTLC estimation of bioactive compound tetrandrine was highest in ACP228.4 mg/mg followed by CP-29.62 mg/mg, CCP-23.46 mg/mg, MCP-18.82 mg/mg and HCP-1.25 mg/mg. TEThas been isolated from ACP.Conclusion: The results of the present in vitro assays revealed that the alkaloid fraction (ACP) is the mostactive fraction when compared to other extracts and has a positive correlation with the levels of bioactivecompound tetrandrine.

Curcumin Reverses The Scopolomine-Induced Cognitive Impairment By Inos And Acetylcholinesterase Inhibition In Rodent Model Of Schizophrenia

Schizophrenia is a severe neuro-developmental psychiatric disorder. Curcumin is a polyphenolic compound extracted from turmeric. It is known for its antioxidant, anti-inflammatory, neuroprotective, and precognitive properties. The purpose of the current study was to evaluate the role of curcumin in scopolamine induced cognitive impairment in animal model of schizophrenia. The elevated plus-maze test was utilised to study the curcumin effect on learning and memory. Curcumin (100 mg/kg, i.p.) was administered daily for 28 days in animals. Behavioural tests such as transfer latency (TL) and spontaneous alteration behaviour was assessed after the last dose of curcumin on the 28th day, followed by biochemical estimations. Present study reported that curcumin showed anti-amnesic effect in animal models of cognitive impairment of schizophrenia. Curcumin reduced the TL compared to toxic control group (scopolamine per se) (P <0.001) in elevated plus maze. In spontaneous alteration behaviour test, curcumin significantly increased percentage alteration and possible alteration as compared to toxic control group (P <0.001). A significant change in acetyl cholinesterase activity, nitrate and oxidative parameters was observed, thus, confirming its anti acetyl cholinesterase, NOS (nitric oxide synthase) inhibition and antioxidant properties (P <0.05). The present study put forward the claim of curcumin as a new and safer therapeutic alternative for the treatment of cognitive impairment in Schizophrenia. The underlying mechanism of this potential effect may be related to anticholinesterase and nitric oxide synthase inhibition activity of curcumin. Further research is warranted for confirming the suggested pathways accountable for memory alleviating effects of curcumin in Schizophrenia.

Effect of Betulic Acid on RAGE/NF-κB/iNOS Signaling Pathway Expression in Steatosis Cell / 中国实验方剂学杂志

Objective::To investigate the effect of betulic acid(BA) on steatosis LO2 cells. Method::LO2 cells were intervened with BA at different gradient concentrations (0, 10, 20, 40, 80, 160, 250 μmol·L-1) for 24 hours. methyl thiazolyl tetrazolium(MTT) staining was used to observed cell viability to determine the final concentration of BA. The cells were divided into control, model, dimethylsulfoxide (DMSO) and BA groups, as well as BA groups intervened with low, middle and high concentrations. First, model, DMSO and BA group's cells were cultured in 10% Lipid Mix 1 medium for 24 hours to establish a nonalcoholic fatty liver model. Then, DMSO group and low, medium and high-concentration groups were separately cultured with 0.1%DMSO medium and 20, 40, 80 μmol·L-1 BA medium for 24 hours. And control and model groups were cultured in drug-free medium for 24 hours. Oil red O staining and Nile red staining were used to observe the intracellular lipid droplets. Immunofluorescence was used to detect the protein expression of inducible nitric oxide synthase (iNOS). Western blot was used to detect the protein expression levels of receptor for advanced glycation end-products (RAGE), nuclear factor κB p65 (NF-κB p55) and iNOS. Result::BA within the concentration of 80 μmol·L-1 had no significant toxicity on LO2 cells. Compared with control group, the intracellular lipid droplets were significantly increased in the model group, and the expressions of oxidative stress-related proteins RAGE, NF-κB p65 and iNOS also increased significantly(P<0.05). Compared with model group, the intracellular lipid droplets in DMSO group were similar to those in model group, with no significant difference in the three protein expressions between the two groups. However, the intracellular lipid droplets deposition in the BA group was significantly decreased. And the expressions of RAGE, NF-κB p65 and iNOS proteins in high-concentration BA group were significantly decreased(P<0.05, P<0.01). Conclusion::BA can significantly improve the intracellular fat deposition in LO2 cells, which was probably related to the inhibition of the expressions of oxidative stress-related proteins RAGE, NF-κB p65 and iNOS.

Combination Artesunate and Tinospora crispa Decreases Ubiquitin, HIF-1α, VEGF and iNOS Expression in Brain of Cerebral Malaria Mice Model

Introduction: Cerebral malaria is the most severe complication of Plasmodium falciparum infection. The pathophysiology of cerebral malaria is still unclear, but it is expected caused by cytoadherence, rosetting, autoagglutinations and abundant pro-inflammatory response that will induce the release of secondary molecules like ubiquitin, HIF-1, VEGF, and iNOS. Aim: To determine the effect of Artesunate and brotowali extract (Tinospora crispa) against the expression of ubiquitin, HIF-1α, VEGF and iNOS in the brain of cerebral malaria mice model. Methods: An experimental study of post-test only control group using CB57BL/6J mice model malaria had been done. Samples were divided into 7 groups: negative control (K-), positive control (K+), Artesunate 32 mg/BWkg/ day (P1), Tinospora crispa 70 mg/BWkg/day (P2), combination Artesunate and Tinospora crispa dose 50 mg/ kgBW (P3), combination Artesunate and Tinospora crispa dose 60 mg/kgBW (P4) and combination Artesunate and Tinospora crispa dose 70 mg/kgBW (P5). Mice model were decapitated at 7th day after infection. Expression of ubiquitin, HIF-1α, VEGF, and iNOS was measured by immunohistochemistry. Result: One Way ANOVA showed different expression of ubiquitin, HIF-1α, VEGF and iNOS among groups. Tukey test showed, there was no significant difference in expression of ubiquitin, VEGF and iNOS among single therapy (Artesunate or Tinospora crispa) with combination therapy (p>0.05). Expression of HIF-1α were significantly different between single therapy (Artesunate or Tinospora crispa) with combination therapy of Artesunate and Tinospora crispa 60 mg/kgBW (p=0.019, p=0.013) and combination therapy of Artesunate and Tinospora crispa 70 mg/kgBW (p=0.034; p=0.023). Pearson correlation showed negative correlation between Tinospora crispa dose and expression of HIF-1α (p=0.001; r=-0.832) and iNOS (p=0.001, r=-0.874). Conclusion: The combination of Artesunate and brotowali (Tinospora crispa) extract generally decreases Ubiquitin, HIF-1 α, VEGF dan iNOS expression of cerebral malaria model although only brain HIF-1α expression gives significant decreasing.

Evaluation of Antidiabetic Potentiality of Truffles and Balanites Aegyptiaca among Streptozotocin Induced Diabetic Rats

The potential application of Truffle (Terfeziaceae) and Desert Date tree (Balanites aegyptiaca) for modulating the diabetesmellitus related symptoms has gained much interest. However, less firm evidence has come from data to increase theunderstanding of the mechanism by which Truffle and Balanites protect pancreatic β-cells. The present study aimed toevaluate the effect of methanolic extract of Truffle and Balanites aegyptiaca on the fasting blood glucose (FBG) level; thechanges in pancreatic histology as well as the changes in iNOS and IL-1β genes expression level among STZ induceddiabetic rats which might help in better clarification of possible mechanisms beside the beneficial effects of the studiedplants on diabetes. The elevation of the FBG level, pathological pancreatic changes and the level of both IL-1β and iNOSgene expressions in diabetic rats were observed in comparison with negative control rats. This increase was declinedsignificantly due to the administration of Truffle and Balanites extracts. All of the studied parameters did not completelyreverse to the normal levels as compared with negative control rats. The obtained results concluded that the beneficial effectof Truffle and Balanites aegyptiaca on STZ-induced diabetes was at least partly due to the reduction of IL-1β and iNOS geneover the expression which can have a protective effect on β cell.

Advances of nitric oxide and inducible nitric oxide synthase in asthma / 国际儿科学杂志

Asthma is a chronic inflammatory respiratory disease characterized by variable airway obstruction, airway hyperresponsiveness, and airway remodeling.In the respiratory tract, nitric oxide(NO) is generated via oxidation of L-arginine that is catalyzed by inducible nitric oxide synthase(iNOS). In hypoxia-ischemia and inflammation processes, iNOS is induced to produce excessive NO, playing an important role in asthma, including the modulation of airway and vascular smooth muscle tone and the inflammation.This review focuses on recent research achievements regarding the relationship between NO and iNOS and asthma.

Expression of Krupple-like factor 6 and inducible nitric oxide synthase in apoptosis of RAW264.7 infected by pseudomonas aeruginosa supernatant / 中国药理学通报

Aim To detect the expression of Krupple-like factor 6 ( KLF6) and inducible nitric oxide syn-thase (iNOS) in mouse mononuclear macrophage leukemia cells infected with pseudomonas aeruginosa (PA) supernatant, and to investigate the role of KLF6 protein and iNOS protein in the apoptosis of RAW264. 7 infected with PA supernatant. Methods The effect of PA supernatant on the proliferation of RAW264. 7 was detected by MTT assay. The apoptotic rate was detected by flow cytometry. The morphological changes of RAW264.7 were detected by Hoechst 33342 staining. The effect of PA supernatant on KLF6 and iNOS protein expression was detected by Western blot. Results PA supernatant inhibited the proliferation of RAW264.7 in a concentration- A nd time-depend-ent manner. PA supernatant significantly increased the percentage of RAW264.7 apoptosis and promoted the expression of KLF6 and iNOS. Conclusions PA can inhibit the proliferation of RAW264. 7, which may induce apoptosis through KLF6 and iNOS protein expres-sion.

Inhibitory functions of SaaB on LPS-induced RAW264. 7 cells via NF-κB-iNOS-NO signaling pathway / 中国药理学通报

Aim: To investigate the functional influences of saponins from Anemarrhena asphdeloids Bge (SAaB) on lipopolysaccharide (LPS)-induced RAW264. 7 cells and the regulation of SAaB to NF-κB- iNOS-NO signaling pathway. Methods: The inflammatory cell model in vitro was established using LPS- induced RAW264. 7 cells, and the levels of NO, iN- OS, TNF-α and IL-6 in inflammatory RAW264. 7 cells effected by SAaB were analyzed using Griess and ELISA method respectively. The protein expression levels of NF-κB p65 were measured by Western blot. Results: Compared with control group, LPS (10 mg L-1) -induced RAW264. 7 cells expressed a significant increase in the levels of NO, iNOS, TNF-α, IL-6 and NF-κB p65 protein. SAaB (0.3, 3, 30 mg L-1) obviously reduced the contents of these inflammatory factors (P <0. 01) and the expression of NF-κB p65 protein (P < 0. 01) in LPS-induced RAW264. 7 cells. Conclusion: SAaB can inhibit the functions of LPS-induced RAW264. 7 cells by modulating NF-KB- iNOS-NO signaling pathway.

Advances of nitric oxide and inducible nitric oxide synthase in asthma / 国际儿科学杂志

Asthma is a chronic inflammatory respiratory disease characterized by variable airway ob-struction, airway hyperresponsiveness, and airway remodeling. In the respiratory tract, nitric oxide ( NO) is generated via oxidation of L-arginine that is catalyzed by inducible nitric oxide synthase ( iNOS) . In hypoxia-ischemia and inflammation processes, iNOS is induced to produce excessive NO, playing an important role in asthma, including the modulation of airway and vascular smooth muscle tone and the inflammation. This re-view focuses on recent research achievements regarding the relationship between NO and iNOS and asthma.

Protective effect of luteolin on acute lung injury in sepsis mice / 中华急诊医学杂志

Objective To investigate the potential therapeutic effect of luteolin on sepsis-induced ALI and the underlying mechanisms.Methods Total of 50 mice were randomly(random number) divided into five groups:a sham control group,a sepsis-induced ALI group,and three sepsis groups pre-treated with 20,40,and 80 mg/kg body weight luteolin.Mice in the treatment groups were pre-treated with luteolin at the respective oral dose two days before ALI induction.The lungs were isolated for histopathological examinations,and the bronchoalveolar lavage fluid (BALF) was collected for biochemical analyses.Results Luteolin significantly attenuated sepsis-induced ALI.Additionally,luteolin treatment decreased protein and inflammatory cytokine concentration and the number of infiltrated inflammatory cells in BALF compared with that in the non-treated sepsis mice.Pulmonary myeloperoxidase (MPO) activity was lower in the luteolin-pre-treated sepsis groups than in the sepsis group.The mechanism underlying the protective effect of luteolin on sepsis is related to the up-regulation of certain antioxidation genes,including inducible nitric oxide synthase (iNOS),cyclooxygenase-2 (COX-2),superoxide dismutases (SODs),and heme oxygenase 1 (HO-1),and the reduction of inflammatory responses through blockage of the activation of the nuclear factor (NF)-κB pathway.Conclusions Luteolin pre-treatment inhibits sepsis-induced ALI through its anti-inflammatory and antioxidative activity,suggesting that luteolin may be a potential therapeutic agent for sepsis-induced ALI.

Thai pigmented rice bran extracts inhibit production of superoxide, nitric oxide radicals and inducible nitric oxide synthase in cellular models

Objective: To study the inhibitory effect of rice bran extracts of Thai black Kam Muang and red Hawm Dawk Mali Deang on oxidative stress factors including superoxide (O2?-), nitric oxide (NO?), and inducible nitric oxide synthase (iNOS). Methods: Bran extracts (40% ethanol) of Kam Muang and Hawm Dawk Mali Deang were obtained and evaluated for in vitro 2-2′-azino-di-(3-ethylbenzthiazoline sulfonate) (ABTS) and NO? scavenging activity. Their inhibitory effects on cellular O2?- and NO? were measured in phorbol 12-myristate 13-acetate-stimulated neutrophil-like HL-60 cells and lipopolysaccharide-stimulated RAW264.7 macrophages, respectively, and their viability was monitored using the MTT assay. The effect on iNOS expression was also assessed by the Western blotting assay. Total contents of phenolics, flavonoids, and subtypes were also determined. Results: Hawm Dawk Mali Deang exhibited about 3.5-fold greater cellular O2?- inhibitory activity than Kam Muang [EC50 values of (23.57±4.54) and (81.98±1.45) μg/mL, respectively] in phorbol 12-myristate 13-acetate-stimulated HL-60 cells. Hawm Dawk Mali Deang exhibited about 2-fold higher in vitro ABTS?+ and NO? scavenging activity than Kam Muang, but it exerted cellular NO? inhibitory activity of only about 26% (undetermined EC50 value) in lipopolysaccharide-stimulated RAW264.7 cells. Conversely, Kam Muang exerted potent cellular NO? inhibitory activity [EC50 value: (281.13±59.18) μg/mL] and dose-dependently decreased iNOS levels. No cytotoxicity of both extracts was detected in both cell types. As for corresponding contents, Hawm Dawk Mali Deang contained higher contents of phenolics and flavonoids than Kam Muang. Moreover, Kam Muang and Hawm Dawk Mali Deang had a high content of total anthocyanins [(14.73±0.52) mg C3GE/g of extract] and total proanthocyanidins [(115.13±1.47) mg CE/g of extract], respectively. Conclusions: Based on these data, bran extracts of Thai black Kam Muang and red rice Hawm Dawk Mali Deang can help lower oxidative stress and inflammation attributed partly to O2?-and NO?.

Thai pigmented rice bran extracts inhibit production of superoxide, nitric oxide radicals and inducible nitric oxide synthase in cellular models

Objective: To study the inhibitory effect of rice bran extracts of Thai black Kam Muang and red Hawm Dawk Mali Deang on oxidative stress factors including superoxide (O

Two new ursane-type nortriterpenes from Lonicera macranthoides and their iNOS-inhibitory activities / 中国天然药物

The flower buds of Lonicera macranthoides (Shan Yin-Hua), represent an important traditional Chinese medicine and food ingredient. A phytochemical investigation of the 70% EtOH extract of the flower buds of L. macranthoides resulted in the isolation of 12 triterpenoids (1-12), including two new ursane-type nortriterpenes, 2α, 24-dihydroxy-23-nor-ursolic acid (1) and 2α, 4α-dihydroxy-23-nor-ursolic acid (2). Their structures were established by multiple spectroscopic methods and comparison with literature data. All isolated compounds were evaluated for their anti-inflammatory effects in LPS-activated RAW264.7 cells. Compounds 1 and 2 exhibited inhibitory effects on iNOS at the concentration of 30 μmol·L.