Pim-2减轻H_2O_2诱导乳鼠心肌细胞损伤及三七皂苷R1的干预研究

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目的:研究Pim-2基因在过氧化氢(H_2O_2)诱导乳鼠心肌细胞损伤中的作用及三七皂苷R1的干预效应。方法:H_2O_2干预原代培养的乳鼠心肌细胞造成细胞损伤模型,分别以三七皂苷R1和Pim-2 RNAi干预细胞,MTT法检测细胞增殖,流式细胞术测细胞凋亡,实时定量PCR法测细胞内Pim-2 mRNA水平,Western blotting法测Pim-2、Bax及Bcl-2蛋白表达。结果:H_2O_2上调Pim-2 mRNA及蛋白水平,抑制细胞增殖与促进凋亡,与对照组相比差异有统计学意义(P<0.001)。Pim-2基因沉默后,细胞活性降低,凋亡增加,LDH与MDA浓度增加,SOD活性降低,Bax蛋白表达增加,Bcl-2蛋白水平降低,与对照组相比差异有统计学意义(P<0.001)。三七皂苷R1预处理能显著增加细胞活力,抑制细胞凋亡,上调Pim-2蛋白水平,与H_2O_2组相比差异有统计学意义(P<0.001)。结论:Pim-2能减轻H_2O_2致心肌细胞损伤,可能与抗凋亡,抗氧化,调控凋亡相关蛋白Bax及Bcl-2表达有关;三七皂苷R1抗心肌细胞损伤与Pim-2有关。 AIM: To investigate the role of Pim-2 in H 2 O 2 -induced cardiomyocyte injury in neonatal rat cardiomyocytes and the effect of notoginsenoside-R1 on it. Methods: Primary cultured neonatal rat cardiomyocytes were treated with H 2 O 2 to induce cell injury. The cells were exposed to notoginsenoside R 1 and Pim-2 RNAi respectively. Cell proliferation was measured by MTT assay. Apoptosis was measured by flow cytometry. The level of Pim-2 mRNA in cells was measured. The expressions of Pim-2, Bax and Bcl-2 were detected by Western blotting. Results: H 2 O 2 increased Pim-2 mRNA and protein levels, inhibited cell proliferation and promoted apoptosis. Compared with the control group, the difference was statistically significant (P <0.001). After silencing Pim-2, cell viability decreased, apoptosis increased, LDH and MDA concentrations increased, SOD activity decreased, Bax protein expression increased, and Bcl-2 protein level decreased. The difference was statistically significant compared with the control group (P < 0.001). Pretreatment with notoginsenoside R1 significantly increased cell viability, inhibited apoptosis and up-regulated Pim-2 protein levels compared with H 2 O 2 group (P <0.001). Conclusion: Pim-2 can reduce H 2 O 2 -induced injury of cardiomyocytes, which may be related to anti-apoptosis, anti-oxidation and regulation of apoptosis-related proteins Bax and Bcl-2. Pim-2 may play an important role in cardiomyocyte injury.
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