目的 研究丹参配方颗粒对乙醇损伤大鼠肾上腺嗜铬细胞瘤(PC12)细胞的保护作用及其机制。方法 用乙醇和不同质量浓度丹参配方颗粒溶液处理PC12细胞;MTT法检测细胞存活率, 乳酸脱氢酶(LDH)法检测丹参配方颗粒对乙醇致PC12细胞损伤的保护作用;应用原位末端转移酶标记法(TUNEL)和Annexin V/PI标记染色检测PC12细胞凋亡情况;相关试剂盒测定总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化氢酶(GSH-Px)的活性和丙二醛(MDA)的量;DCFH-DA染色法检测胞内活性氧(ROS)水平。结果 丹参配方颗粒能够抑制乙醇导致的PC12细胞的损伤和死亡, 显著降低乙醇诱导的PC12细胞的凋亡率(P < 0.05), 提高细胞的T-SOD、CAT和GSH-Px酶活性(P < 0.05), 降低MDA和ROS水平(P < 0.05)。结论 丹参配方颗粒能有效保护乙醇损伤的PC12细胞, 其作用机制与提高细胞抗氧化能力有关。

Objective To investigate the protective effects of Danshen Formula Granula (DFG) against the damage of pheochromocytoma (PC12) cells induced by alcohol. Methods PC12 cells were treated with alcohol and DFG at different concentration. The survival rates of PC12 cells were measured by MTT method and the protective effects of DFG against the damage of PC12 cells induced by alcohol were evaluated using the lactate dehydrogenase (LDH) leakage assays (P < 0.05). The apoptosis rates in PC12 cells were examined by TUNEL and Annexin V/PI label staining (P < 0.05). The antioxidant mechanism was assayed by detecting the activities of total superoxide dismutase (T-SOD), catalase (CAT), and glutathione peroxide enzyme (GSH-Px), and the levels of malondialdehyde (MDA) and intracellular reactive oxygen species (ROS) (P < 0.05) were detected by DCFH-DA staining method. Results DFG could reduce the damage of PC12 cells induced by alcohol and significantly decrease the apoptosis rate induced by alcoholic damage in PC12 cells (P < 0.05). Meanwhile, DFG could increase the activity of T-SOD, GSH-Px, while CAT, the amounts of MDA, and intracellular ROS were decreased. Conclusion DFG has the effective protection against the damaged PC12 cells induced by alcohol. The mechanism may be involved in increasing the cell anti-oxidative function.

国家自然科学基金面上项目(30670688);河南省卫生厅AIDS科技攻关项目(2006037);河南省卫生厅科技攻关项目(200803085);河南省教育厅自然科学研究项目(2009A330001)