Inhibitory Effects Of Hedyotis Diffusa And Scutellaria Barbata On The Proliferation Of High Glucose-induced HBZY-1 Cells And Its Mechanism

Objective: To study the effects of Hedyotis diffusa(HD)and Scutellaria barbata(SB)alone or in combination with each other on the proliferation of HBZY-1cells induced by high glucose and its mechanism.Methods: Sulforhodamine B(SRB)assay was used to detect the proliferation of HBZY-1 cells.The activities of SOD,CAT and GSH-Px enzyme and the content of MDA and NO in HBZY-1 cells were detected by their kits.The level of ROS in HBZY-1 cells was determined by DCFH-DA fluorescence probe.The expression levels of matrix metalloproteinases MMP-2 and MMP-9 were detected by Gelatin zymogram assay.Western blotting method was used to detect N-cadherin,Vimentin,Erk1/2,p-Erk1/2,P38,p-P38,Akt,p-Akt,IKK and p-IKK prot-ein expression in HBZY-1 cells.Results: The inhibitory effects of HD and SB on the growth of HBZY-1 cells was weak,and the inhibitory rates of the highest concentration of 400 μg/ml for 72 h were29.57% and 43.80%,respectively.30 mM glucose(HG)significantly promoted the proliferation of HBZY-1 cells for 24 h,and the proliferation rate was 127.39%.The activities of intracellular antioxidant enzyme SOD,CAT and GSH-Px were decreased,but the content of MDA,NO and ROS were increased.HD and SB alone significantly inhibited the growth of HBZY-1 cells stimulated by HG,and 400 μg/ml of HD and SB completely inhibited the growth of HBZY-1 cells stimulated by HG.The combination of HD and SB at 100 μg/ml significantly increased the activities of SOD,CAT and GSH-Px and decreased the production of MDA,NO and ROS induced by HG,which was significantly stronger than that of HD and SB 200 μg/ml alone.HG significantly decreased the expression of MMP-2 and MMP-9 and increasedthe expression of N-cadherin and Vimentin.The combination of HD and SB at 100μg/ml significantly increased the expression of MMP-2 and MMP-9 and decreased the expression of N-cadherin and Vimentin induced by HG.The effect was significantly stronger than that of HD and SB 200 μg/ml alone.HG also increased the protein expressions of p-Erk,p-P38,p-Akt and p-IKK in HBZY-1 cells.The combination of HD and SB at 100 μg/ml significantly inhibited the expression of p-Erk,p-P38 and p-IKK,and decreased the ratios of p-Erk/Erk,p-P38/P38,and p-IKK/IKK.The effects were significantly stronger than those of HD and SB 200 μg/ml alone.Conclusion: the combination of HD and SB could significantly inhibit the proliferation of HBZY-1 cells induced by high glucose,and the mechanism is related to the enhancement of antioxidant capacity and inhibition of MAPK and NF-kappa B signaling pathway in HBZY-1 cells.