Protective Effect of Dihuang Yinzi on Cerebral Ischemia-reperfusion Injury in Rats and Its Mechanism / 中国实验方剂学杂志

Objective: To explore the protective effect and the preliminary mechanism of Dihuang Yinzi on cerebral ischemia-reperfusion injury in rats. Method: The middle cerebral artery occlusion (MCAO) model was established. Totally 90 SD rats were randomly divided into 6 groups:sham operation group, model group, nimodipine group (0.01 g·kg-1) and high, medium, low-dose Dihuang Yinzi groups (38.80, 19.40, 9.70 g·kg-1), with 20 rats in each group.The modified neurological severity score (mNSS) was assayed at the 7th, 14th, 28th days after operation, and the volume of cerebral infarction, pathological changes of brain tissue, the BrdU positive cells and mRNA levels of Notch1, Jagged1 and Hes1 in subventricular zone(SVZ)were observed respectively by triphenyl tetrazolium chloride(TTC) stain, htorylin eastin(HE) stain, immunofluorescence technique and reverse transcriphase polymerase chain reaction(Real-time PCR) methods at the 28th day after the operation. Result: The mNSS on the 7th, 14th, 28th days of high, medium-dose Dihuang Yinzi groups and nimodipine group were significantly lower than that of model group(PPth day, the percentage of cerebral infarction volume in brain tissue volume of high, medium-dose Dihuang Yinzi groups and nimodipine group were smaller than that of model group(Pth day, the BrdU positive cells in SVZ of the above 3 groups were significantly higher than model group(PPPth day, the mRNA levels of Notch1, Jagged1 and Hes1 of high, medium-dose Dihuang Yinzi groups and nimodipine group were significantly higher than those of model group(PPPPConclusion: Dihuang Yinzi can improve the nerve function defect of MCAO rat model, and reduce the volume of cerebral infarction and the pathological changes of brain tissue, thus playing a protective role in cerebral ischemia-reperfusion injury rats. Its mechanism may be related to the activation of the Notch signaling pathway, and the up-regulation of expressions of Notch1, Jagged1 and Hes1 mRNA, thus promoting the proliferation of NSCs.

Effect of Panax Notoginseng Saponins on EMT of Rat Renal Proximal Tubular Epithelial Cells Induced by TGF-β1 / 中国实验方剂学杂志

Objective:To investigate the intervention effect of panax notoginseng saponins (PNS) on epithelial-mesenchymal transition (EMT) of rat renal tubular epithelial cells (NRK-52E) induced by transforming growth factor-β1(TGF-β1), and analyze the mechanism based on the silent information regulation 2 homolog 1(SIRT1)/TGF-β1/Smad signaling pathway. Method:NRK-52E were cultured in DMEM medium with 10% fetal bovine serum, and divided into normal control group, TGF-β1 group (5 μg·L-1), resveratrol (RSV) group (50 mg·L-1), EX527 group (10 μmol·L-1), Panax notoginseng saponins (PNS) group (100 mg·L-1), and EX527+ PNS group (10 μmol·L-1+100 mg·L-1). Then cells were collected after drug intervention for 48 h. The expressions of α-SMA,E-cadherin,SIRT1,TGF-β1,Smad3,Smad4 mRNA in each group were detected by Real-time PCR. The protein expressions of α-SMA, E-cadherin,SIRT1 and TGF-β1 were detected by Western blot. Result:Compared with normal group, mRNA and protein expressions of α-SMA increased obviously(PPβ1 group. Compared with TGF-β1 group, mRNA and protein expressions of α-SMA decreased significantly(PPPβ1,Smad3,and Smad4 decreased(PConclusion:PNS can prevent the occurrence of EMT of renal tubular epithelial cells induced by TGF-β1, and the mechanism may be related to active SIRT1 to inhibit TGF-β1/Smad pathway.

Influence of Tangshenfang on diabetic liver injury and the expression of SIRT1 and PGC-1αin the liver tissue / 中国病理生理杂志

AIM:To observe the effect of Tangshenfang(TS)on the liver protection and the levels of silent in-formation regulator 1(SIRT1)and peroxisom proliferator-activated receptor γcoactivator-1α(PGC-1α)in the liver tissue. METHODS:The rat model of diabetes mellitus(DM)was established by intravenous injection of streptozotocin(STZ;30 mg/kg)after having the high fat/high glucose diets for 1 month.The diabetic rats were randomly divided into DM group,DM with high-dose TS(TSHi)group, medium-dose TS(TSMed)group and low-dose TS(TSLow)group.The normal rats were served as control group.There were 8 rats in each group.After treatment with TS for 12 weeks,the serum biochemical indi-ces including fasting blood glucose(FBG), triglyceride(TG), alanine aminotransferase(ALT)and aspartate aminotrans-ferase(AST)were tested.Fasting insulin(FINS)was also detected by radioimmunoassay,and homeostatic model assess-ment for insulin resistance(HOMA-IR)was calculated.The serum levels of tumor necrosis factor-α(TNF-α)and interleu-kin-1(IL-1)were measured by ELISA.The activity of SOD and content of MDA in the liver tissues were measured by the methods of hydroxylamine and thiobarbituric acid.The liver pathological changes were observed under light microscope with HE and Masson staining.The protein expression of SIRT1and PGC-1αin the liver tissues was determined by Western blot. RESULTS:In DM group,serum FBG,TG,ALT,AST,FINS,HOMA-IR,TNF-αand IL-1 were obviously increased com-pared with the control group(P<0.01).The fatty changes,local necrosis,inflammation and fibrosis in the liver tissues were observed.The content of MDA in liver increased,while the activity of SOD decreased markedly.The protein expression of SIRT1 and PGC-1αwas decreased(P<0.05).In TS treatment groups,all these changes in DM rats were markedly reversed by TS,and the protein expression of SIRT1 and PGC-1αin the liver tissues was markedly increased.CONCLUSION:TS may protect the rats from diabetic liver injury by increasing the expression of SIRT 1 and PGC-1α,and thereby improving in-sulin resistance and oxidative stress.

Panax notoginseng saponins protect kidney from diabetes by up-regulating silent information regulator 1 and activating antioxidant proteins in rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To explore the mechanism of the protective effects of Panax notoginseng saponins (PNS) on kidney in diabetic rats.</p><p><b>METHODS</b>Diabetic rat model was obtained by intravenous injection of alloxan, and the rats were divided into model, PNS-100 mg/(kg day) and PNS-200 mg/(kg day) groups, 10 each. Another 10 rats injected with saline were served as control. Periodic acid-Schiff staining and immunological histological chemistry were used to observe histomorphology and tissue expression of bone morphogenetic protein-7 (BMP-7). Silent information regulator 1 (SIRT1) was silenced in rat mesangial cells by RNA interference. The mRNA expressions of SIRT-1, monocyte chemoattractant protein-1 (MCP-1), transforming growth factor β1 (TGF-β1) and plasminogen activator inhibitor-1 (PAI-1) were analyzed by reverse transcription polymerase chain reaction. The protein expressions of SIRT1 and the acetylation of nuclear factor κB (NF-κB) P65 were determined by western blotting. The concentration of MCP-1, TGF-β1 and malondialdehyde (MDA) in culture supernatant were detected by enzyme-linked immuno sorbent assay. The activity of superoxide dismutase (SOD) was detected by the classical method of nitrogen and blue four.</p><p><b>RESULTS</b>In diabetic model rats, PNS could not only reduce blood glucose and lipid (P<0.01), but also increase protein level of BMP-7 and inhibit PAI-1 expression for suppressing fibrosis of the kidney. In rat mesangial cells, PNS could up-regulate the expression of SIRT1 (P<0.01) and in turn suppress the transcription of TGF-β1 (P<0.05) and MCP-1 (P<0.05). PNS could also reverse the increased acetylation of NF-κB p65 by high glucose. In addition, redox regulation factor MDA was down-regulated (P<0.05) and SOD was up-regulated (P<0.01), which were both induced by SIRT1 up-regulation.</p><p><b>CONCLUSIONS</b>PNS could protect kidney from diabetes with the possible mechanism of up-regulating SIRT1, therefore inhibiting inflammation through decreasing the induction of inflammatory cytokines and TGF-β1, as well as activating antioxidant proteins.</p>

Effect of SIRT1 on high glucose-induced NF-κB p65 subunit acetylation and MCP-1 expression in rat mesangial cells / 第二军医大学学报

Objective: To explore the effect of silent information regulator 1(SIRT1) on high glucose-induced nuclear factor-κB (NF-κB) p65 subunit acetylation and monocyte chemoattractant protein 1 (MCP-1) expression in rat mesangial cells (RMCs). Methods: The lentiviral shRNA plasmid pTRC-shSIRT1 was constructed for interference of SIRT1 gene and was identified. The RMCs were divided into high glucose group (treated with high glucose culture medium), resveratrol (SIRT1 activator)+high glucose group (treated with low glucose culture medium containing 1 μmol/L resveratrol for 24 h, and then with high glucose culture medium), SIRT1 RNAi group (4 h after viral pTRC-shSIRT1 infection, and then treated with low-glucose culture medium), SIRT1 RNAi + high glucose group (4 h after viral pTRC-shSIRT1 infection, and then treated with high glucose culture medium); we also established normal control group and hypertonic mannitol control group. The mRNA expression of SIRT1 and MCP-1 gene was analyzed by real-time quantitative PCR; the protein expression of SIRT1 and the acetylation of NF-κB p65 subunit were observed by Western blotting analysis. The protein level of MCP-1 in the supernatants was detected by ELISA. Results: DNA sequencing confirmed the successful construction of the plasmid pTRC-shSIRT1, which could knocked down SIRT1 mRNA expression(P<0.01). High glucose decreased SIRT1 expression and promoted acetylation of NF-κB p65 subunit, and increased MCP-1 mRNA and protein expression. Resveratrol, an activator of SIRT1, could reverse the above changes induced by high glucose. Conversely, silencing SIRT1 gene significantly accelerated the high glucose-induced acetylation of NF-κB p65 subunit and MCP-1 expression at both mRNA and protein levels(P<0.01 or P<0.05). Conclusion: SIRT1 can inhibit high glucose-induced MCP-1 mRNA and protein expression in RMCs, which may involve NF-κB p65 deacetylation.

Oxidized low density lipoprotein-induced expression of transforming growth factor beta 1 and production of extracellular matrix in rat glomerular mesangial cells is mediated by lectin-like oxidized low density lipoprotein receptor-1 / 第二军医大学学报

Objective: To investigate the effect of oxidized low-density lipoprotein (ox-LDL) on expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), transforming growth factor beta 1 (TGF-β 1), and secretion of extracelluar matrix (ECM) in cultured rat glomerular mesangial cells (GMCs), and to investigate the influence of LOX-1 inhibitor polyinosinic acid (PIA) on the effect of ox-LDL. Methods: Rat glomerular masengial cells were cultured in vitro. RT-PCR was employed to determine the LOX-1 mRNA expression in GMCs incubated with different concentrations of ox-LDL (0, 25, 50, 100 μg/ ml). The expression of LOX-1 and TGF-β1, mRNA was also determined by RT-PCR in the blank control group, ox-LDL (50 μg/ml) group and PIA(50 μg/ml ox-LDL+250 μg/ml PIA) group. The contents of TGF-β1 fibronectin (FN), and collagen IV (Col IV) in the supernatants of the above 3 groups were determined by ELISA. Results: RT-PCR showed that LOX-1 mRNA expression in 25, 50 and 100 μg/ml ox - LDL groups was significantly higher than that of blank control group(P<0.05), with the highest expression found in the 50 μg/ml ox-LDL group; the expression of LOX-I and TGF-β 1 mRNA was significantly higher in 50 μg/ml ox-LDL group than that in the other 2 groups (P< 0.01). ELISA results demonstrated that the supernatant contents of TGF-β1, FN and Col IV were significantly higher in 50 μg/ml ox-LDL group than those in the other 2 groups (P< 0.05). Conclusion: Ox-LDL can upregulate the expression of LOX-1 and TGF-β1, mRNA and the secretion of extracelluar matrix in GMCs. Polyinosinic acid can antagonize the above effect of ox-LDL, suggesting that LOX-1 may participate in ox-LDL-induced GMCs damage and is involved in the development and progression of glomerulosclerosis.

Experiment study of tongfu huoxue decoction in the treatment of intracelebral hemorrhage / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effects of Tongfu Huoxue decoction on experimental intracelebral hemorrhage and the associated machenisms.</p><p><b>METHOD</b>The cerebral hemorrhage model in rats was induced by local injection of type VII collagenase and they were randomly divided into four groups. The treated groups were treated with Naoxuekang and Tongfu Huoxue decoction. The control groups were only treated with water. The changes of neurological defect were observed. The content of brain water, MDA, NO and the activity of SOD were measured.</p><p><b>RESULT</b>The cerebral hemorrhage rats showed hemiplegia, and the hemorrhage brains showed celebral edema, higher quotient of brain and content of brain water, suggesting the hemorrhage model was established successfully. After the treatment of Tongfu Huoxue decoction, the hemorrhage rats showed smaller hemorrhage volume, the brain tissue from the hemorrhage rats had lower MDA content and the quotient of brain, and also had higher activity of SOD and content of NO.</p><p><b>CONCLUSION</b>Tongfu Huoxue decoction has treatment effects on cerebral hemorrhage.</p>