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目的研究葛根素注射液对人近曲小管上皮细胞HK-2细胞株增殖的影响,以及对高糖诱导后细胞内78KD-糖调节蛋白(GRP78)表达的影响。方法设置葛根素质量浓度梯度,依次为5、10、20、40、80、160、320、640 g/L,采用MTT方法,检测HK-2细胞的增殖情况。HK-2细胞传代贴壁后,无血清培养24 h后,分为正常组、模型组和葛根素治疗组,继续培养48 h后,提取各组细胞总RNA,采用Real Time PCR的方法,检测GRP78 mRNA的表达情况。结果随着浓度的增大,葛根素注射液对HK-2细胞增殖的抑制作用增强。经葛根素注射液作用48 h后,与正常组相比,模型组GRP78 mRNA的表达量增多,差异有显著性(P<0.01);与模型组比较,治疗组GRP78 mRNA的表达量增加,差异有显著性(P<0.01)。结论葛根素注射液对HK-2细胞增殖具有抑制作用;高糖刺激HK-2细胞48 h后,可导致其发生内质网应激反应,葛根素注射液可通过增加内质网伴侣蛋白GRP78的表达量,以促进蛋白质正确折叠,来恢复细胞的稳态。
Objective To study the effects of puerarin on the proliferation of human proximal tubule epithelial cells HK-2 and its effect on the expression of 78 kD-glycoprotein (GRP78) after induced by high glucose. Methods The concentration gradient of puerarin was set as 5, 10, 20, 40, 80, 160, 320 and 640 g / L respectively. The proliferation of HK-2 cells was detected by MTT assay. After subculture, the cells were divided into normal group, model group and puerarin-treated group. After cultured for 48 h, the total RNA of each group was extracted. Real Time PCR was used to detect the expression of HK-2. GRP78 mRNA expression. Results With the increase of concentration, the inhibitory effect of puerarin on HK-2 cell proliferation increased. Compared with the normal group, the expression of GRP78 mRNA in the model group increased significantly after 48 h of puerarin injection (P <0.01). Compared with the model group, the GRP78 mRNA expression in the treatment group increased There was significant (P <0.01). CONCLUSION Puerarin injection can inhibit the proliferation of HK-2 cells. After high glucose stimulation of HK-2 cells for 48 h, ER stress can induce endoplasmic reticulum stress. Puerarin injection can increase the endoplasmic reticulum protein GRP78 The amount of expression, to promote the correct protein folding, to restore cell homeostasis.