灯盏花素保护大鼠免受糖尿病肾病侵害的作用机制研究  

Mechanism of Breviscapine Against Diabetic Nephropathy in Rats

作　　者：王静[1] 李敏[1] 何立明[1] 张志佳 檀增桓[1] WANG Jing;LI Min;HE Liming;ZHANG Zhijia;TAN Zenghuan(Department of Gastroenterology,Handan Central Hospital,Handan 056000,China)

机构地区：[1]邯郸市中心医院内分泌科,邯郸056000

出　　处：《世界中医药》2022年第22期3162-3167,共6页World Chinese Medicine

基　　金：河北省卫生和计划生育委员会医学科学研究课题计划项目(20181668)。

摘　　要：目的:探究灯盏花素改善大鼠糖尿病肾脏侵害的作用机制。方法:将24只Wistar大鼠分为对照组(n=6)、模型组(n=9)和灯盏花素干预组(n=9)。通过链脲佐菌素诱导构建1型糖尿病Wistar大鼠模型。通过免疫比浊法检测血液糖化血红蛋白水平,结合尿液样本中白蛋白/肌酐比值,超氧化物歧化酶和丙二醛酶联免疫吸附试验检测结果,评估大鼠肾脏损伤情况。采用苏木精-伊红(HE)染色和透射电子显微镜观察大鼠肾皮质组织肾小球病理变化。使用TUNEL测定法检测肾脏细胞凋亡情况;通过蛋白质印迹法分析p-AMP活化蛋白激酶、p-糖原合酶激酶-3β(GSK-3β)、BCL-2-相关X蛋白(BAX)和裂解胱天蛋白酶-3(Cleaved Caspase-3)蛋白表达水平。结果:与模型组比较,灯盏花素干预组大鼠的血糖水平及糖化血红蛋白含量均明显降低(P<0.05);肾小球损伤和肾脏组织病变显著减轻(P<0.05);大鼠尿液中超氧化物歧化酶(SOD)水平显著升高,丙二醛水平显著降低(P<0.05);核因子E2相关因子2(Nrf2)/Keap1蛋白比值和血红素加氧酶-1(HO-1)蛋白水平均明显升高(P<0.05)。与模型组比较,灯盏花素干预组大鼠肾皮质组织中AMP活化蛋白激酶和乙酰辅酶A羧化酶(ACC)蛋白磷酸化水平显著上调,固醇调节元件结合蛋白-1(SREBP-1)、SREBP-2和脂肪分化相关蛋白(ADRP)表达水平均显著下调(P<0.05)。与模型组比较,灯盏花素干预组大鼠肾脏组织细胞凋亡减少,p-GSK-3β、BAX和Cleaved Caspase-3蛋白表达水平显著降低(P<0.05)。结论:灯盏花素通过降低大鼠血糖水平,激活AMPK和Nrf2信号通路抑制肾脏脂质积累和氧化应激,激活AKT信号通路抑制高血糖刺激诱导的细胞凋亡,从而发挥肾脏保护作用。Objective:To explore the mechanism of breviscapine in improving diabetic nephropathy in rats.Methods:Twenty-four Wistar rats were divided into a control group(n=6),a model group(n=9),and a breviscapine intervention group(n=9).Streptozocin was used to induce the type 1 diabetes model in Wistar rats.The level of glycosylated hemoglobin in the blood was detected by immunoturbidimetry.The renal injury in rats was evaluated by the albumin/creatinine ratio,superoxide dismutase(SOD),and malondialdehyde(MDA)detected by enzyme-linked immunosorbent assay(ELISA).H&E staining and transmission electron microscopy(TEM)were used to examine the pathological changes of glomeruli in the renal cortex of rats.Renal cell apoptosis was detected using TUNEL assay.The protein expression levels of phosphorylated adenosine 5′-monophosphate-activated protein kinase(p-AMPK),phosphorylated glycogen synthase kinase-3β(p-GSK-3β),Bcl-2-associated X protein(BAX),and cleaved caspase-3 were detected by Western blot.Results:Compared with the model group,the breviscapine intervention group showed reduced blood glucose level and glycosylated hemoglobin content(P<0.05),alleviated glomerular injury and renal tissue lesions(P<0.05),increased SOD level in the urine,reduced MDA level(P<0.05),and elevated nuclear factor E2-related factor 2(Nrf2)/Keap1 protein ratio and heme oxygenase-1(HO-1)protein level(P<0.05).Furthermore,the breviscapine intervention group also showed up-regulated p-AMPK and phosphorylated acetyl-CoA carboxylase(ACC)protein expression in the renal cortical tissues,down-regulated protein expression levels of sterol regulatory element binding protein(SREBP-1),SREBP-2,and adipose differentiation-related protein(ADRP)(P<0.05),reduced apoptosis of renal tissues,and decreased protein expression levels of p-GSK-3β,BAX,and cleaved caspase-3.Conclusion:Breviscapine can reduce the renal blood glucose level,activate AMPK and Nrf2 signaling pathways to inhibit renal lipid accumulation and oxidative stress,and activate the AKT signaling pathway

关 键 词：灯盏花素 链脲佐菌素 糖尿病肾病 氧化应激 细胞凋亡 AMPK信号通路 Nrf2信号通路 AKT信号通路 

分 类 号：R587.1[医药卫生—内分泌]