五味子乙素对高糖诱导的PC12细胞损伤的保护作用

The protective effect of Schisandrin B on PC12 cells damaged by high glucose

摘要

目的 探讨五味子乙素（Sch B）对高糖（HG）引起的大鼠肾上腺嗜铬细胞瘤（PC12）细胞损伤的保护作用并探讨其可能机制。方法 采用MTT法筛选可诱导PC12细胞损伤的HG适宜浓度与处理时长;将不同浓度Sch B作用于PC12 细胞,筛选出适宜的药物浓度。利用HG构建糖尿病认知功能障碍体外模型,加入Sch B后再次检测细胞活力、细胞凋亡情况和细胞活性氧（ROS）水平。试剂盒检测培养基中的乳酸脱氢酶（LDH）的释放量,以评价细胞的损伤程度。试剂盒检测细胞中超氧化物歧化酶（SOD）活力和丙二醛（MDA）含量,以评价氧化应激程度。结果 本研究筛选确定125 mmol·L^-1葡萄糖干预PC12细胞48 h可构建稳定HG损伤细胞模型。与正常对照组相比,HG处理48 h后PC12细胞活力显著降低（P<0.01）,细胞凋亡率、LDH释放量、胞内氧化应激水平均明显升高;1~20 μmol·L^-1 Sch B无细胞毒性,且可呈剂量依赖性拮抗HG诱导的细胞损伤,20 μmol·L^-1 Sch B可显著抑制细胞凋亡、减少LDH释放量、下调ROS蓄积并缓解氧化应激紊乱（P<0.01）。结论 Sch B可以改善HG对PC12细胞的损伤作用,调节氧化应激,降低细胞凋亡,从而改善糖尿病认知功能障碍。

Abstract

Objective To investigate the protective effect of Schisandrin B (Sch B) against high glucose (HG)-induced cytotoxicity in rat adrenal pheochromocytoma (PC12) cells and elucidate its underlying mechanisms. Methods MTT assay was adopted to screen the optimal concentration and treatment duration of HG for inducing PC12 cell injury. PC12 cells were treated with different concentrations of Sch B to screen non-cytotoxic drug concentrations. An in vitro model of diabetic cognitive dysfunction was established by HG stimulation. After Sch B intervention, cell viability, cell apoptosis and intracellular reactive oxygen species (ROS) levels were detected. The release of lactate dehydrogenase (LDH) in culture medium was determined by commercial kits to evaluate the degree of cell injury. The activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in cells were measured to assess the level of oxidative stress. Results This study confirmed that treatment of PC12 cells with 125 mmol·L^-1 glucose for 48 h could establish a stable HG-induced cell injury model. Compared with the normal control group, 48 h of HG exposure markedly reduced PC12 cell viability (P<0.01), while the apoptosis rate, LDH release and intracellular oxidative stress level were significantly elevated. Sch B at concentrations ranging from 1 to 20 μmol·L^-1 exhibited no cytotoxicity and alleviated HG-triggered cell injury in a dose-dependent manner. Specifically, 20 μmol·L^-1 Sch B significantly suppressed cell apoptosis, decreased LDH leakage, reduced ROS accumulation and restored disturbed oxidative stress balance (P<0.01). Conclusion Sch B can ameliorate the damage of HG to PC12 cells, regulate oxidative stress, reduce cell apoptosis, and thus improve diabetic cognitive impairment.

关键词

五味子乙素 / 糖尿病认知功能障碍 / PC12细胞 / 氧化应激 / 细胞凋亡 / 神经保护

Key words

Schisandrin B / diabetic cognitive dysfunction / PC12 cell / oxidative stress / cell apoptosis / neuroprotection

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