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目的:研究艳山姜挥发油对氧化低密度脂蛋白(ox-LDL)诱导人主动脉内皮细胞(HAECs)损伤的保护作用,并分析可能的作用机制。方法:实验分为空白组(无血清内皮细胞培养基),模型组(200 mg·L-1ox-LDL),艳山姜挥发油高剂量组(200mg·L-1ox-LDL+100μg·L-1艳山姜挥发油)和低剂量组(200 mg·L-1ox-LDL+10μg·L-1艳山姜挥发油)共4组。艳山姜挥发油预保护1 h,继续加入ox-LDL共同作用24 h,采用噻唑盐(MTT)法分析细胞存活率,苏木精-伊红(HE)染色法进行形态学观察;采用酶联免疫吸附法(ELISA)检测培养上清液乳酸脱氢酶(LDH)外漏量,Western blot分析c-Jun氨基末端激酶(JNK)1/2/3,p-JNK1/2/3蛋白表达,实时荧光定量聚合酶链式反应分析JNK1/2/3 mRNA表达水平。结果:艳山姜挥发油能显著提高HAECs的存活率,改善细胞的病理损伤,减少LDH外漏量,抑制JNK1/2/3蛋白磷酸化,但对JNK1/2/3蛋白和mRNA水平无显著影响。结论:艳山姜挥发油改善ox-LDL诱导的HAECs损伤,其作用机制与抑制JNK1/2/3蛋白磷酸化相关。
Objective: To study the protective effect of volatile oil from Yammycin Ginkgo against oxidative low density lipoprotein (ox-LDL) -induced injury of human aortic endothelial cells (HAECs) and to analyze its possible mechanism. Methods: The experiment was divided into blank group (serum-free endothelial cell culture medium), model group (200 mg · L-1 ox-LDL), Yanshan Ginger volatile oil high dose group (200 mg · L -1 ox-LDL + 100 μg · L -1 Yam ginger volatile oil) and low-dose group (200 mg · L-1ox-LDL + 10μg · L-1 yam ginger volatile oil) a total of 4 groups. Yanshan ginger volatile oil pre-protection 1 h, ox-LDL continue to join 24 h, cell viability was analyzed by MTT method, the morphology was observed by hematoxylin-eosin (HE) staining; The amount of lactate dehydrogenase (LDH) leakage in the culture supernatant was detected by ELISA. The protein expression of c-Jun N-terminal kinase (JNK) 1/2 and p-JNK1 / 2 / Real-time fluorescence quantitative polymerase chain reaction analysis of JNK1 / 2/3 mRNA expression levels. Results: Volatile oil from Yam Ginger significantly enhanced the survival rate of HAECs, improved the pathological damage of cells, reduced the leakage of LDH and inhibited the phosphorylation of JNK1 / 2/3, but had no significant effect on the protein and mRNA level of JNK1 / 2/3 . CONCLUSION: Volatile oil from Yammycin increases the injury of HAECs induced by ox-LDL, and its mechanism of action is related to the inhibition of JNK1 / 2/3 protein phosphorylation.