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Objective:To investigate the protective effect of quercetin （Qu） on articular cartilage of knee osteoarthritis and its mechanism by inhibiting p38 mitogen activated protein kinase （MAPK） signaling pathway. Method:Through the network pharmacology technology，we scientifically predicted and analyzed the target factors and signal pathways of Qu in the protection of articular cartilage in patients with osteoarthritis. We selected a prediction pathway closely related to osteoarthritis and validated it by cell experiment <italic>in vitro</italic>. The best intervention concentration of the drug was selected by cell counting kit-8 （CCK-8） method. The osteoarthritis and its closely related inflammatory factors interleukin（IL）-1<italic>β</italic> and tumor necrosis factor（TNF）-<italic>α</italic> were detected by enzyme linked immunosorbent assay（ELISA）. The expression of related mRNA and protein in p38 signal pathway after Qu intervention were detected by quantitative real time polymerase chain reaction（Real-time PCR） and Western blot. Result:It was predicted that MAPK signal pathway was closely related to osteoarthritis by network pharmacology，and p38 MAPK pathway，which was most closely related to osteoarthritis，was selected for study. The results showed that 100 μmol·L<sup>-1</sup> Qu had the most obvious effect in decreasing the expression of related inflammatory factors，inhibited the expression of p38，phosphorylated（p）-p38，matrix metalloproteinase-13（MMP-13），A disintegrin-like and metalloproteinase with thrombospondin type-1 motifs-4（ADAMTS-4） in the pathway，and promoted the expression of CollagenⅡ. Conclusion:Qu could decrease the expression of cartilage inflammatory factors in the prevention and treatment of osteoarthritis，and the effect can be well developed by intervening and inhibiting p38 MAPK pathway related factor expression level. All the results show that Qu can decrease osteoarthritis inflammatory factors and protect articular cartilage in patients with osteoarthritis.
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Objective:To investigate the mechanism of joint cavity injection of Dioscoreae Rhizoma polysaccharides （DRP） in protecting against cartilage degeneration and inhibiting the expression of inflammatory factors in the rabbit model of knee osteoarthritis to provide relevant references for the development and further research on DRP. Method:Fifty-five New Zealand white rabbits were selected for the induction of knee osteoarthritis model by the modified Hulth's modeling method. The model rabbits were randomly divided into a model group， a sodium hyaluronate group （1.00 mg·kg<sup>-1</sup>）， and low- （0.7 mg·kg<sup>-1</sup>）， medium- （1.43 mg·kg<sup>-1</sup>）， and high-dose （2.15 mg·kg<sup>-1</sup>） DRP group according to a random number table. One week after modeling， the rabbits in the groups with drug intervention were treated correspondingly for five weeks， once per week， and no intervention was performed in the model group. Five weeks later， the joint specimens were observed by visual observation. The articular cartilage tissues were observed under the light microscope for pathological sections and scores by hematoxylin-eosin （HE） staining and toluidine blue （TB） staining. The expression levels of interleukin-6 （IL-6）， interleukin-1<italic>β</italic> （IL-1<italic>β</italic>）， and tumor necrosis factor-<italic>α </italic>（TNF-<italic>α</italic>） in the synovial fluid were measured by enzyme-linked immunosorbent assay （ELISA）， and the expression of matrix metalloproteinase-13 （MMP-13）， transforming growth factor-<italic>β</italic><sub>1</sub> （TGF-<italic>β</italic><sub>1</sub>）， and type Ⅱ collagen （Col-Ⅱ） in the articular cartilage were measured by immunohistochemistry. Result:After five weeks of DRP intervention， compared with the model group， the DRP groups exhibited lowered levels of IL-6， IL-1<italic>β</italic>， and TNF-<italic>α</italic> in the synovial fluid （<italic>P</italic><0.05）， reduced expression of MMP-13 in the articular cartilage （<italic>P</italic><0.05）， and increased levels of TGF-<italic>β</italic><sub>1 </sub>and Col-Ⅱ （<italic>P</italic><0.05）. Compared with the low-dose and high-dose DRP groups， the medium-dose DRP group showed reduced levels of IL-6， IL-1<italic>β</italic>， and TNF-<italic>α</italic> in the knee joint （<italic>P</italic><0.05）， increased levels of TGF-<italic>β</italic><sub>1</sub> and Col-Ⅱ in cartilage tissues （<italic>P</italic><0.05）， and dwindled level of MMP-13 （<italic>P</italic><0.05）. Compared with the sodium hyaluronate group， the medium-dose DRP group showed no significant differences in IL-6， IL-1<italic>β</italic>， and TNF-<italic>α</italic> in rabbit knee joints， and TGF-<italic>β</italic><sub>1</sub>， Col-Ⅱ， and MMP-13 in cartilage tissues. Conclusion:Joint cavity injection of DRP can significantly reduce the expression of IL-6， IL-1<italic>β</italic>， and TNF-<italic>α</italic> in rabbit synovial fluid， effectively inhibit the expression of MMP-13 in the articular cartilage to suppress the degradation of articular cartilage collagen and promote the synthesis of TGF-<italic>β</italic><sub>1</sub> and Col-Ⅱ. Therefore， DRP can protect and repair articular cartilage to delay the degeneration of articular cartilage.
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Objective To expore the impact of teniary butyl hydroquinone (tBHQ) on Müller cells in SD rats retina under high glucose condition,and discuss the mechanism of tBHQ.Methods The Müller cells of SD rats were cultured in vitro and the experiment was divided into normal control group,high glucose group and tBHQ intervention group.Western blot and quantitative real time PCR were used to determine the expression of nuclear factor erythroid 2-related factor 2 (Nrf2),hemeoxygenase-1 (HO-1),hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) protein and mRNA in each group.Results Müller cells cultured in vitro were flat with irregularly sharp.The nucleus was oval,while the cytoplasm was abundant.Adjacent cells were interwoven to a network.Western blot assay showed the overall expression of Nrf2,HO-1,HIF-1α,and VEGF in Müller cells of normal control group,high glucose group,and tBHQ intervention group were significantly different (F =73.831,148.618,152.269,91.217,all at P＜0.001);Among them,the relative expressions of Nrf2,HO-1,HIF-1α and VEGF proteins in the high glucose group were 0.17±0.02,0.47±0.02,0.67±0.07,and 0.6±0.05,which were increased in comparion with 0.06±0.01,0.19±0.03,0.06±0.00 and 0.07±0.02 in the normal control group,with statistically significant differences (t =4.114,9.275,16.479,13.353,all at P ＜ 0.001);the relative expressions of Nrf2 and HO-1 proteins in the tBHQ intervention group were 0.40±0.06 and 0.72±0.05,which were increased by comparion with those in the higher glucose group,with statistically significant differences (t =7.847,7.947,both at P＜0.001);the relative expressions of HIF-1α and VEGF proteins in the tBHQ intervention group were 0.18±0.04,0.26±0.07,which were decreased in comparion with those in the higher glucose group,but were increased in comparion with those in normal control group,with statistically significant differences (t =13.215,8.444,both at P＜ 0.001).Quantitative real time PCR showed that the relative mRNA expressions of Nrf2,HO-1,HIF-1α,and VEGF in Müller cells of normal control group,high glucose group,and tBHQ intervention group were significantly different (F =340.317,1 582.911,488.852,185.699,all at P＜0.001);the relative mRNA expressions of Nrf2,HO-1,HIF-1α,and VEGF proteins in the high-glucose group were 1.53 ± 0.06,1.50 ± 0.04,2.56 ± 0.09,and 3.04 ± 0.19,which were increased in comparion with 1.07±0.07,0.95±0.05,0.99±0.02,and 1.09±0.08 in the normal control group,with statistically significant differences (t =7.292,15.014,30.550,18.573,all at P ＜ 0.001);The relative mRNA expressions of Nrf2 and HO-1 proteins in the tBHQ intervention group were 2.68±0.09 and 2.94±0.05,which were increased in comparion with those in the high-glucose group,with statistically significant differences (t =18.046,39.458,both at P＜0.001);The relative mRNA expression of HIF-1α and VEGF protein in the tBHQ intervention group were 1.48±0.05 and 1.6±0.08,which were decreased by comparion with those in the higher glucose group were increased in comparion with those in normal control group,with statistically significant differences (t =21.036,13.739,both at P＜0.001).Conclusions tBHQ protects Müller cells from damage in high glucose condition by activating anti-oxidative stress signaling pathway of Nrf2/ARE.
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Objective@#To expore the impact of teniary butyl hydroquinone (tBHQ) on Müller cells in SD rats retina under high glucose condition, and discuss the mechanism of tBHQ.@*Methods@#The Müller cells of SD rats were cultured in vitro and the experiment was divided into normal control group, high glucose group and tBHQ intervention group.Western blot and quantitative real time PCR were used to determine the expression of nuclear factor erythroid 2-related factor 2 (Nrf2), hemeoxygenase-1 (HO-1), hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) protein and mRNA in each group.@*Results@#Müller cells cultured in vitro were flat with irregularly sharp.The nucleus was oval, while the cytoplasm was abundant.Adjacent cells were interwoven to a network.Western blot assay showed the overall expression of Nrf2, HO-1, HIF-1α, and VEGF in Müller cells of normal control group, high glucose group, and tBHQ intervention group were significantly different (F=73.831, 148.618, 152.269, 91.217, all at P<0.001); Among them, the relative expressions of Nrf2, HO-1, HIF-1α and VEGF proteins in the high glucose group were 0.17±0.02, 0.47±0.02, 0.67±0.07, and 0.6±0.05, which were increased in comparion with 0.06±0.01, 0.19±0.03, 0.06±0.00 and 0.07±0.02 in the normal control group, with statistically significant differences (t=4.114, 9.275, 16.479, 13.353, all at P<0.001); the relative expressions of Nrf2 and HO-1 proteins in the tBHQ intervention group were 0.40±0.06 and 0.72±0.05, which were increased by comparion with those in the higher glucose group, with statistically significant differences (t=7.847, 7.947, both at P<0.001); the relative expressions of HIF-1α and VEGF proteins in the tBHQ intervention group were 0.18±0.04, 0.26±0.07, which were decreased in comparion with those in the higher glucose group, but were increased in comparion with those in normal control group, with statistically significant differences (t=13.215, 8.444, both at P<0.001). Quantitative real time PCR showed that the relative mRNA expressions of Nrf2, HO-1, HIF-1α, and VEGF in Müller cells of normal control group, high glucose group, and tBHQ intervention group were significantly different (F=340.317, 1 582.911, 488.852, 185.699, all at P<0.001); the relative mRNA expressions of Nrf2, HO-1, HIF-1α, and VEGF proteins in the high-glucose group were 1.53±0.06, 1.50±0.04, 2.56±0.09, and 3.04±0.19, which were increased in comparion with 1.07±0.07, 0.95±0.05, 0.99±0.02, and 1.09±0.08 in the normal control group, with statistically significant differences (t=7.292, 15.014, 30.550, 18.573, all at P<0.001); The relative mRNA expressions of Nrf2 and HO-1 proteins in the tBHQ intervention group were 2.68±0.09 and 2.94±0.05, which were increased in comparion with those in the high-glucose group, with statistically significant differences (t=18.046, 39.458, both at P<0.001); The relative mRNA expression of HIF-1α and VEGF protein in the tBHQ intervention group were 1.48±0.05 and 1.6±0.08, which were decreased by comparion with those in the higher glucose group were increased in comparion with those in normal control group, with statistically significant differences (t=21.036, 13.739, both at P<0.001).@*Conclusions@#tBHQ protects Müller cells from damage in high glucose condition by activating anti-oxidative stress signaling pathway of Nrf2/ARE.
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OBJECTIVE: To investigate the protective effect and possible mechanism of Rhizoma Coptis(RC) on lipopolysaccharide(LPS)-induced inflammatory injury in rat hepatocytes(BRL). METHODS: LPS-induced BRL cells injury model was established in vitro, then the damaged cells were given different interventions and treatment with 0.175, 0.1 mg• mL-1 RC aqueous extract as the test drug, and dexamethasone(Dex) as positive control drug. The optimal test doses of LPS and RC aqueous extract were selected and determined by cell counting kit-8(CCK-8), the cellular apoptosis rate was determined by flow cytometry, TLR4/NF-κB and TLR4/IRF3 signaling pathways and the mRNA level of related inflammatory mediators(TNF-α, IL-1β, IL-6) were detected by RT-PCR, the NF-κB p65 protein expression was analysed by Western blot and immunofluorescence techniques. RESULTS: ①Compared with normal control group, 0.1 mg•mL-1 LPS affected on BRL cells for 24 h, the cell survival rate was decreased significantly(P<0.01), the apoptotic rate increased significantly(P<0.01), the mRNA level of TLR4, NF-κB, IRF3, TNF-α, IL-1β, IL-6 were significantly increased(P<0.01), and the NF-κB p65 protein expression was increased. ②Compared with the model group, 0.1 and 0.175 mg•mL-1 RC affected on LPS-induced BRL cells for 24 h, the survival rate of BRL cells was increased significantly(P<0.05), the apoptotic rate decreased significantly(P<0.01), the mRNA level of TLR4, NF-κB, IRF3, TNF-α, IL-1β, IL-6 and the NF-κB p65 protein expression were decreased significantly(P<0.01). CONCLUSION: Rhizoma Coptis has obviously protective effect on LPS-induced inflammatory injury in rat hepatocytes(BRL), the mechanism of which may be related with inhibiting apoptosis, reducing the release of inflammatory factors such as TNF-α, IL-1β and IL-6, blocking NF-κB p65 protein nuclear translocation, interfering the R4/NF-κB and TLR4/IRF3 signaling pathway.
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Diabetic kidney disease is a common complication in patients with diabetes mellitus and so far,lacking effective therapy aiming at it.Glucagon like peptide-1 (GLP-1) receptor agonist is a novel hypoglycemic agent associated with incretin.Studies have shown that GLP-1 have additional renal protection independent of its hypoglycemic effect,mainly through the mechanisms such as antihypertension,reducing glomerular hyperfiltration,lowering proteinuria,anti-inflammatory,antioxidative stress and anti-RASS.In this review,we will summary their renal effects and mechanisms from the basic and clinical evidences.
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OBJECTIVE:To investigate the protective effect of Compound ginkgo leaf granules on liver function damage of acute pancreatitis. METHODS:Totally 82 patients with severe acute pancreatitis(SAP)complicated with liver function damage se-lected from our hospital during Sept. 2013-Sept. 2015 were randomized into control group and observation group according to ran-dom number table,with 41 cases in each group. Control group was given SAP related routine therapy. On the basis of the control group,observation group was additionally given Compound ginkgo leaf granules 5 g added into warm boiled water 80 mL via naso-gastric tube,tid,one week as a treatment course,for 2 courses. The remission time of abdominal pain,serum amylase recovery time,length of hospital stay and adverse drug reactions were observed in 2 groups. The levels of serum amylase,ALT,TBIL, AST,γ-GT,ALB,IL-6,TNF-α,creotoxin concentration,MDA,SOD,GSH-Px,total antioxidant capacity(TAC)and peripher-al blood PMN NF-κB levels were determined in 2 groups before and after treatment. RESULTS:In control group,3 patients with-drew from the study and 38 patients completed the study;in observation group,6 patients withdrew from the study and 35 patients completed the study. The remission time of abdominal pain,recovery time of serum amylase and the length of hospital stay in obser-vation group were significantly shorter than control group,with statistical significance (P0.05). After treatment,the levels of serum amylase,ALT,TBIL, AST,γ-GT,IL-6,TNF-α,creotoxin concentration,NF-κB and MDA in 2 groups were decreased significantly,while the levels of ALB,SOD,GSH-Px and TAC were increased significantly;the observation group was significantly lower or higher than the con-trol group,with statistically significant (P0.05). CONCLUSIONS:Compound ginkgo leaf granules can significantly decrease serum amylase,protect liver function, with good safety. The mechanismmay be inhibiting the release of inflammatory factors,improving oxidative stress reactions,reduce the concentration of creotoxin and inhibiting the activity of NF-κB.
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Panax notoginsenosidum(PNS), which was proved to have the efficacy of increasing cerebral blood flow,improving microcirculation and anti-platelet aggregation, has been widely used for the treatment of cerebral ischemia-reperfusion injury(CIRI). A line of current studies have shown that the protective mechanism of PNS on CIRI is associated with multiple ways, including the roles of PNS on free radicals and lipid peroxides, apoptosis, calcium channels, neuronal damage, and the protein expression of signaling pathways etc. The above protective mechanisms underlying the role of PNS on CIRI provide well-founded scientific evidences for the exploitation and application of PNS. In this paper, the progress in protective mechanism of PNS on CIRI by summarizing and analyzing data collected from literatures published in recent years is reviewed.
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lschemic postconditioning(IPo)is a way that after long ischemia on liver graft,animals are given one or several brief reperfusion-ischemia before persistent reperfusion to improve the hepatic tolerance and relieve the ischemie reperfusion injury.It has been proved an effective and controlled method to attenuate the ischemic reperfusion injury.Protective mechanism of ischemic postconditioning on hepatic graft is related with protecting sinus hepaticus endotheliocyte and hepatic microcirculation,relieving hepatic cells injury and inflammatory reaction induced by oxygen free radicals,relieving calcium ovedoad in hepatic cells and mitochondria,regulating apoptosis genes,transforming ion channels condition in mitochondria.This article will makes a brief review on protective mechanism ofhepatic ischemic posteanditioning.