补肾活血方对高糖高脂诱导损伤的小鼠脑微血管内皮细胞内质网应激和RAGE/LRP-1蛋白表达的影响  

Effects of Bushen Huoxue Formula on the protein expression of endoplasmic reticulum stress and RAGE/LRP-1 in BEND3 cells damaged by high glucose and high palmitate

作　　者：闫敬来 张益铭 赵佳奇 田雅娟 李钦青 贺文彬 YAN Jing-lai;ZHANG Yi-ming;ZHAO Jia-qi;TIAN Ya-juan;LI Qin-qing;HE Wen-bin(Shanxi Key Laboratory of Chinese Medicine Encephalopathy,ShanxiUniversity of Chinese Medicine,Jinzhong 030619,China)

机构地区：[1]山西中医药大学中医脑病学山西省重点实验室,晋中030619

出　　处：《中华中医药杂志》2021年第1期397-401,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金重点项目(No.81530099),国家自然科学基金面上项目(No.81473375);山西省回国留学人员科研资助项目(No.2013-134)。

摘　　要：目的:在细胞水平探索补肾活血方对高糖高脂诱导的小鼠脑微血管内皮细胞(BEND3细胞)损伤的改善作用,并探究补肾活血方对高糖高脂导致的内质网应激和转运Aβ相关受体蛋白的作用。方法:采用高糖高脂(25mmol/L葡萄糖,200μmol/L棕榈酸)刺激BEND3细胞,用CCK-8法筛选合适的补肾方(BS)、活血方(HX)、补肾活血方(BSHX)的最佳用药浓度;应用免疫蛋白印迹法检测BS、HX、BSHX最佳用药浓度对葡萄糖调节蛋白78(GRP78)、磷酸化真核起始因子2α(p-eIF2α)、活性转录因子4(ATF4)、核因子κB(NF-κB)抗体以及脑内与转运β淀粉样蛋白相关的晚期糖基化终产物受体(RAGE)和低密度脂蛋白受体相关蛋白-1(LRP-1)的表达情况。结果:CCK-8检测细胞活力,最终选定补肾方100mg/L、活血方20mg/L和补肾活血方120mg/L;与正常对照组比较,高糖高脂刺激BEND3细胞后,GRP78、p-eIF2α、ATF4、NF-κB、RAGE表达显著增加(P<0.01),LRP-1表达下降(P<0.01);药物干预后,GRP78、p-eIF2α、ATF4、NF-κB、RAGE蛋白下调,BSHX组LRP-1表达显著上调(P<0.05,P<0.01);且以BSHX组最为明显。结论:补肾方、活血方、补肾活血方对高糖高脂诱导的BEND3细胞损伤均有改善作用,但以补肾活血方疗效最为突出,且对高糖高脂导致的内质网应激和转运β淀粉样蛋白受体的蛋白表达具有调节作用。Objective:To investigate the effects and mechanisms of Bushen Huoxue Formula against high glucoseinduced damage in mouse brain microvascular endothelial cells(BEND3 cells).Methods:In vitro experiments using high glucose(25 mmol/L,HG)and palmitic acid(200μmol/L,PA)intervention BEND3 cells with CCK-8 method to screen the best concentration of Bushen Formula,Huoxue Formula and Bushen Huoxue Formula.The expression of glucose-regulated protein 78(GRP78),phosphorylated eukaryotic initiation factor 2α(p-eIF2α),active transcription factor 4(ATF4),nuclear factor kappa-B(NF-κB),advanced glycosylated end-product receptor(RAGE)and low-density lipoprotein receptor-associated protein-1(LRP-1),was determined by Western Blot assay at the optimal concentration of Bushen Formula,Huoxue Formula and Bushen Huoxue Formula.Results:CCK-8 was used to detect cell proliferation,and the concentration of Bushen Formula,Huoxue Formula and Bushen Huoxue Formula was finally selected as 100,20,120 mg/L.Compared with normal control group,HG and PA stimulus after BEND3 cells,GRP78,p-eIF2α,ATF4,NF-κB and RAGE protein expression significantly increased(P<0.01)and the expression of LRP-1 protein significantly decreased(P<0.01).After the intervention of Bushen Formula(100 mg/L),Huoxue Formula(20 mg/L)and Bushen Huoxue Formula(120 mg/L),the GRP78,p-eIF2α,ATF4,NF-κB and RAGE protein were significantly decreased and the LRP-1 protein significantly increased(P<0.05,P<0.01).Conclusion:Bushen Formula,Huoxue Formula and Bushen Huoxue Formula can improve BEND3 cell damage induced by high glucose and high palmitate,and regulate endoplasmic reticulum stress and the protein expression of beta amyloid receptor.

关 键 词：补肾活血方 高糖高脂 小鼠脑微血管内皮细胞 内质网应激 晚期糖基化终产物受体 低密度脂蛋白受体相关蛋白-1 

分 类 号：R285.5[医药卫生—中药学]