银杏叶提取物各组分对高糖培养下系膜细胞增殖及细胞外基质积聚的抑制作用

目的: 探讨银杏叶提取物(GBE)不同组分对高糖培养下系膜细胞增殖及细胞外基质(ECM)积聚的作用,阐明其作用机制。方法: 高糖培养的系膜细胞分别给予GBE、总黄酮、总内酯、总黄酮水解物、槲皮素、山奈酚、异鼠李素、银杏内脂A、银杏内脂B和银杏内脂C,药物浓度分别为0(高糖对照组)、3.125、6.250、12.500、25.000和50.000 mg·L^-1,同时设低糖对照组,MTT法检测各组系膜细胞增殖情况。高糖培养的系膜细胞分别给予GBE、总黄酮、总内酯、银杏内脂A、银杏内脂B和银杏内脂C,药物浓度分别为0(高糖对照组)、0.05、0.50、5.00和50.00 mg·L^-1,同时设低糖对照组,ELISA法检测条件培养基中Ⅳ型胶原(Col Ⅳ)和纤维连接蛋白(FN)水平。结果: MTT检测,与高糖对照组比较, 50.000 mg·L^-1GBE组细胞增殖活性明显降低(P<0.01),25.000 和50.000 mg·L^-1总黄酮组细胞增殖活性明显降低(P<0.05或P<0.01),总黄酮水解物、槲皮素、山奈酚和异鼠李素各浓度组细胞增殖活性均明显降低(P<0.01),50.000 mg·L^-1内脂C组细胞增殖活性明显降低(P<0.05);其他各组细胞增殖活性与高糖对照组比较差异均无统计学意义(P>0.05)。ELISA检测,与高糖对照组比较,5.00和50.00 mg·L^-1GBE 组Col Ⅳ和FN水平均明显降低(P<0.01), 0.50 mg·L^-1GBE组FN水平明显降低(P<0.05),0.50、5.00和50.00 mg·L^-1总黄酮组Col Ⅳ水平降低(P<0.05或P<0.01),50.00 mg·L^-1总内脂组Col Ⅳ和FN水平降低(P<0.05或P<0.01),50.00 mg·L^-1内脂 C 组FN水平降低(P<0.05)。结论: GBE在高浓度时(50.000 mg·L^-1)抑制高糖诱导的肾小球系膜细胞增殖而低浓度(自0.500 mg·L^-1起)时抑制系膜细胞ECM积聚;GBE中总黄酮在抑制细胞增殖和ECM积聚方面起主要作用,总内脂无明显效果;苷元抑制系膜细胞增殖的作用远远强于糖苷。

Inhibitory effects of different Ginkgo biloba leaf extract ingredients on proliferation and ECM accumulation of high glucose-treated mesangial cells

Objective To investigate the inhibitory effects of different Ginkgo biloba leaf extract (GBE) ingredients on the proliferation and extracellular matrix (ECM) accumulation of high glucose-treated mesangial cells,and to clarify their mechanisms. Methods The high glucose-treated mesangial cells (HBZY-1) were given GBE,total flavonoids,total ginkgolide,total flavonoids hydrolyzate,quercetin,kaempferol,isorhamnetin,ginkgolide A,B,and ginkgolide C; the concentrations were 0(high glucose control group),3.125,6.250,12.500,25.000 and 50.000 mg·L^-1,meanwhile low glucose control group was set up. The cell proliferation was detected by MTT assay. The high glucose-treated mesangial cells were given GBE,total flavonoids,total ginkgolide,ginkgolide A,ginkgolide B,and ginkgolide C;the concentrations were 0(high glucose control group),0.05,0.50,5.00,50.00 mg·L^-1,meanwhile low glucose control group was set up. The levels of fibronectin(FN) and type Ⅳ collagen (Col Ⅳ) were detected by ELISA method. Results The MTT results showed that compared with high glucose control group,the proliferation activity of mesangial cells in 50.00 mg·L^-1 GBE group was decreased significantly (P<0.01);the proliferation activities of mesangial cells in 25.00 and 50.00 mg·L^-1 total flavonoids group were decreased significantly(P<0.05 or P<0.01);the proliferation activities of mesangial cells in all quercetin,kaempferol,isorhamnetin and total flavonoids hydrolyzate groups were decreased significantly (P<0.01);the proliferation activity of mesganial cells in 50.00 mg·L^-1 ginkgolide C group was decreased significantly (P<0.01). The ELISA results showed that compared with high glucose control group,the levels of Col Ⅳand FN in 5.00 and 50.00 mg·L^-1 GBE groups were decreased significantly (P<0.01),and the FN level in 0.50 mg·L^-1 GBE group was decreased significantly (P<0.05);the Col Ⅳ levels in 0.50,5.00,and 50.00 mg·L^-1 total flavonoids groups were decreased significantly(P<0.05 or P<0.01);the Col Ⅳand FN levels in all total ginkgolide groups had no obviously differences except 50.00 mg·L^-1ginkgolide C group (P<0.05 or P<0.01);the FN level in 50 mg·L^-1 ginkgolide C group was decreased significantly(P<0.05). Conclusion GBE can inhibit the high glucose-induced proliferation only in the high concentration (50 mg·L^-1) and inhibit the high glucose-induced mesangial cell ECM accumulation in the low concentration (0.50-25.00 mg·L-1);the total flavonoids plays a major role in the inhibition of cell proliferation and ECM accumulation,and the total ginkgolide has no significant effect;the inhibitory effect of aglycones on mesangial cell proliferation is much stronger than that of glucoside.