ABSTRACT
Aim To study the role of marein mediated AMPK signaling pathway in delaying oxidative stress, inflammation and fibrotic protein expression in diabetic nephropathy ( DN ). Methods In vitro diabetic nephropathy model was established by HG + PA induced rat mesangial cells ( HBZY-1 ) , and the cultured HBZY-1 cells were divided into normal control group (NG), HG + PA( GlucoselOO mmol • L'1 + Palmitic acid 250 [imo\ • L'1, HG +PA) model group, HG + PA + marein with different doses of 25 p,mol • L"1, 50 p,mol • L"1, 100 jimol • L"1, and 200 junol • L"1 groups. MTS was used to detect the effect of marein on HBZY-1 cell proliferation, and the optimal concentration was selected. Western blot was used to test the protein expression of NOX4, TGF-fU, MCP-1, a-SMA, FN, Collagen VI. Adenosine monophosphate activated( AMPK) protein kinase family of AMPK7I, p-AMPK a expression were measured. Results Marein inhibited high glucose palmitate-induced proliferation of HBZY-1 cells, down-regulated NOX4, TGF-(31, MCP-1, cx-SMA, FN and Collagen VI expression in model cells. Meanwhile, marein up-regulated both AMPK 7I and p-AMPKa expression. Conclusions Marein may inhibit the HBZY-1 cell proliferation, oxidative stress, inflammation and fibrosis factors expression in HG + PA induced HBZY-1 cell by activating of both AMPK 7I and AMPK signaling pathway, thus delaying renal injury in diabetic nephropathy.
ABSTRACT
[ ABSTRACT] AIM:To explore the effects and mechanism of eleutheroside ( ETS) B or E on the proliferation of HBZY-1 cells treated with high glucose.METHODS:The HBZY-1 cells were cultured under high glucose condition.The 4th generation of HBZY-1 cells was used for determining the optimal cell density, which was consistent with the growth reg-ulation curve of the cells.The cells were divided into 6 groups:low glucose ( LG) group, high glucose ( HG) group, high glucose plus ETS-B/E ( low dose, medium dose and high dose) groups, and high glucose plus losartan ( LTG) group.Af-ter all cells were treated with the corresponding drugs at 24 h, 48 h and 72 h, the inhibitory rate of the proliferation was measured, and the expression of TGF-β1 and PPARγwas detected by immunocytochemistry and Western blotting.RE-SULTS:The best cell density was 2 000 cells/well, which was complied with the basic rules of the cell growth, and high glucose significantly promoted the HBZY-1 cell proliferation.At each time point, the inhibitory effects of ETS-B/E were significantly different between HG group and LTG group on the proliferation of the HBZY-1 cells ( P<0.05) .The expres-sion of TGF-β1 was significantly inhibited, and the expression of PPARγwas significantly promoted by ETS-B/E ( P<0.05).ETS-E showed stronger effect than ETS-B (P<0.05) in a concentration-and time-dependent manner.CONCLU-SION:ETS-B/E significantly inhibits the proliferation of HBZY-1 cells under high glucose condition by decreasing TGF-β1 expression and promoting PPARγexpression.