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目的观察白藜芦醇对db/db小鼠胰岛β细胞的作用并探讨其可能机制。方法取12只db/db小鼠,随机分为糖尿病(DM)组和白藜芦醇干预组,每组各6只,另设对照(Con)组小鼠6只。干预4周后,比较各组OGTT后血糖水平,免疫组织化学分析胰岛中3-磷酸肌醇依赖性蛋白激酶1(PDK-1)蛋白表达量的改变。体外培养小鼠胰岛细胞株MIN6细胞并分为正常葡萄糖(NG)组、高糖(HG)组、高糖+白藜芦醇(HG+Res)组、高糖+白藜芦醇+PI3K特异性抑制剂Wortmannin(HG+Res+Wo)组。Western blot测定4组细胞PDK-1、磷酸化蛋白激酶B(p-AKt)水平,CCK-8法和流式细胞仪检测MIN6细胞增殖、凋亡率;ELISA测定上清液中胰岛素水平。结果与DM组比较,白藜芦醇干预组OGTT血糖曲线下面积下降(P<0.05)、胰岛细胞PDK-1蛋白表达增加[阳性率(0.57±0.32)%vs(1.74±0.47)%,P<0.01];与HG组比较,白藜芦醇干预后MIN6细胞中PDK-1、p-Akt蛋白增加、上清液中胰岛素水平增加[(124.95±5.37)vs(181.19±30.77)pmol/L]、细胞增殖(D值)增加[(0.71±0.10)vs(1.02±0.09)]、凋亡率下降[(19.3±2.1)%vs(11.2±1.4)%](P<0.01);与HG+Res组比较,HG+Res+Wo组MIN6细胞PDK-1表达、Akt磷酸化水平减低,上清液中胰岛素水平及细胞增殖下降、凋亡率增加(P<0.05)。结论白藜芦醇可能通过激活胰岛β细胞PDK-1/Akt通路、改善胰岛β细胞功能。
Objective To observe the effect of resveratrol on islet β-cells of db / db mice and to explore its possible mechanism. Methods Twelve db / db mice were randomly divided into diabetes group and resveratrol intervention group, with 6 mice in each group and 6 mice in control group. After intervention for 4 weeks, the level of blood glucose was compared after each OGTT, and the expression of 3-phosphoinositide-dependent protein kinase 1 (PDK-1) protein in islets was detected by immunohistochemistry. The mouse islet cell line MIN6 cells were cultured in vitro and divided into normal glucose (HG) group, high glucose + HG + Res group, high glucose + resveratrol + PI3K Sex inhibitor Wortmannin (HG + Res + Wo) group. The levels of PDK-1 and phosphorylated protein kinase B (p-AKt) were measured by Western blot. The proliferation and apoptosis of MIN6 cells were detected by CCK-8 assay and flow cytometry. The levels of insulin in supernatant were determined by ELISA. Results Compared with DM group, the area under the curve of OGTT blood glucose decreased (P <0.05) and the expression of PDK-1 protein increased (P <0.05) <0.01]. Compared with HG group, the protein of PDK-1 and p-Akt increased in MIN6 cells after resveratrol treatment and the level of insulin in supernatant increased ([124.95 ± 5.37] vs (181.19 ± 30.77) pmol / L (P <0.01). Compared with HG (P <0.01), the apoptotic rate decreased ([(19.3 ± 2.1) vs (11.2 ± 1.4)%] + Res group, PDK-1 expression and Akt phosphorylation in MIN6 cells in HG + Res + Wo group were decreased, but insulin level and cell proliferation in supernatant decreased and apoptosis rate increased (P <0.05). Conclusion Resveratrol may improve pancreatic β-cell function by activating the PDK-1 / Akt pathway of pancreatic β cells.