吴门调脂颗粒对内皮细胞自噬的干预研究

Intervention of tiaozhi granules on autophagy of HUVECs

目的研究调脂颗粒对人脐静脉内皮细胞(HUVECs)在细胞、蛋白质、分子水平自噬的影响。方法运用血清药理学方法,以调脂颗粒含药血清、6-氨基-3-甲基嘌呤(3-MA)、雷帕霉素(Rapamycin)诱导处理HUVECs,通过MTT比色法检测调脂颗粒含药血清对细胞生长的影响,Western blot检测自噬相关蛋白LC3-Ⅱ的相对表达量变化,免疫荧光法检测细胞内自噬小体的数目变化。结果调脂颗粒含药血清处理HUVECs后,细胞活力无明显改变,自噬相关蛋白LC3-Ⅱ的相对表达上调,自噬小体数量显著增加(P<0.05)。调脂颗粒含药血清联合雷帕霉素处理HUVECs后,细胞活力无明显改变,较雷帕霉素单独处理自噬相关蛋白LC3-Ⅱ的相对表达显著下降,自噬小体数量显著减少(P<0.05或P<0.01)。结论调脂颗粒能抑制内皮细胞的过度自噬,促进内皮细胞适度自噬。

Objective To investigate the effect of tiaozhi granules on the autophagy of HUVECs in the cell,protein and molecular levels.Methods HUVECs were induced by tiaozhi granule medicated serum,6-amino-3-methyl purine（3-MA） and rapamycin.MTT was used to detect the cell viability after 48 hours with tiaozhi granule medicated serum.Western blot was used to detect the expressive level of LC3-Ⅱ.Immunofluorescence was used to detect change in the numbers of autophagosomes.Results After HUVECs was processed by tiaozhi granule medicated serum,the cell viability had not change,the relative expression of autophagy related protein LC3-Ⅱ increased,and the number of autophagosomes increased（P〈0.05）.After HUVECs was processed by tiaozhi granule medicated serum and rapamycin,cell viability had no change,while the relative expression of autophagy related protein LC3-Ⅱ and the number of autophagosomes decreased significantly when compared with the results of intervention with rapamycin only（P〈0.05 or P〈0.01）.Conclusion Tiaozhi granules can inhibit excessive autophagy of endothelial cells and promote moderate autophagy in endothelial cells.