楮实子提取物对衣霉素诱导的细胞ERS的改善作用及其分子机制

Improvement Effect of Chushizi(Fructus Broussonetae) Extract on Endoplasmic Reticulum Stress Cell Model Induced by Tunicamycin and Its Molecular Mechanism

目的:探讨楮实子提取物对衣霉素诱导的细胞内质网应激(ERS)模型的影响及其分子机制。方法:楮实子含药血清干预衣霉素处理的PC12细胞制备内质网应激模型,造模后分为ERS模型组和楮实子高、中、低剂量组。运用CCK8法检测各组内质网应激模型PC12细胞存活率;流式细胞仪检测细胞的凋亡率及细胞周期;利用免疫印迹法检测GRP78及Caspase-12蛋白表达。结果:楮实子含药血清可提高经衣霉素处理后的PC12细胞存活率(P<0.05),楮实子使衣霉素(Tu)诱导的内质网应激细胞的细胞周期中的G2期明显增加(6.9%&19%),ERS模型组细胞凋亡率为8.36%,而楮实子高剂量组细胞凋亡率为4.51%,楮实子可下调凋亡相关蛋白GRP78、Caspase-12的表达,降低内质网应激反应。结论:楮实子可有效缓解衣霉素诱导的细胞毒性,增加细胞周期阻滞,其作用机制可能是通过细胞周期阻滞,在G2期对损伤的细胞进行修复,缓解内质网应激的进一步进展,进而抑制细胞凋亡。

Objective： To investigate the effects of Chushizi （Fructus Broussonetae） on the endoplasmie retic- ulum stress model.tel induced by tunieamycin and its molecular mechanism. Methods： The PCI2 cells treated with tunicamycin were Intervened by serum content of Chushizi （Fructus Broussonetae） to make endoplasmic reticulum stress model. The model was divided into 4 groups, including model group ,high dose group, middle dose group and low dose group. The survival rate of PC12 cells in the endoplasmic reticulum stress model was detected by CCK8 method. The apoptotic rate and cell cycle were detected by flow cytometry. The expression of GRP78 and Caspase-12 protein was detected by imnmnoblotting. Results： The serum content of Chushizi （Fructus Broussone- tae） could increase the survival rate of PCI2 cells treated with tunicamycin （P〈0.05）. The G7 phase of the cell line of endoplasmic retieulum induced by tunicamyein （Tu） increased significantly in Chushizi （Fructus Brous- sonetae） gruup（6.9%＆19%）. The apoptotic rate of the model group was 8.36%, and the Chushizi （Fructus Brous- sonetae） group was significantly reduced to 4.51%. The serum content of Chushizi （Fruetus Broussonetae）e down- regulated the expression of apoptosis-related protein GRP78 and Caspase-12, and decreased the eudoplasmic reticulum stress response. Conclusion： Chushizi （Fruetus Broussonetae） can effectively alleviate the eytotoxicity induced by tunicamycin and increase the cell cycle arrest. The mechanism may be that Chushizi （Fruetus Bruussonetae） can repair the damaged cells in G2 phase by cell cycle arrest, alleviate the further progress of en- doplasmie reticulum stress, and thus inhibit apoptosis.