参附益心方对缺氧条件下大鼠原代心肌细胞线粒体DNA相关基因表达的影响＊

目的 研究参附益心方（SF）对缺氧条件下大鼠原代心肌细胞线粒体DNA相关基因表达的影响。方法 取1~3 d SD新生大鼠,分离、培养、鉴定原代心肌细胞。CCK-8检测SF、辅酶Q10（Coenzyme Q10）最适浓度。实验分为Normal（正常）组、Model（缺氧）组、SFL（参附益心方低剂量）组、SFH（参附益心方高剂量）组、Coenzyme Q10组,后三组在缺氧模型基础上分别给予SF 0.25 mg/ml、SF 0.5 mg/ml、Coenzyme Q10 10 1 × 10^-4 mol/L对细胞进行干预,收集细胞并用相关试剂盒检测ATP含量及乳酸脱氢酶（LDH）含量,RT-PCR检测ND-1、ND-2、COX、tfam基因的表达。结果 与Normal组相比,Model组心肌细胞ATP含量显著下降,LDH含量显著升高,ND-1、ND-2、COX、tfam基因表达显著降低,差异具有统计学意义(P ＜ 0.05)。与Model相比,SFL组LDH含量下降,ND-2、COX基因表达升高,差异具有统计学意义(P ＜ 0.05);SFH组ATP含量升高,LDH含量下降,ND-1、ND-2、COX基因表达升高,差异具有统计学意义(P ＜ 0.05)。结论 参附益心方可上调原代心肌细胞线粒体DNA相关基因COX、ND1、ND2的表达,提高心肌ATP的含量,降低LDH含量,并呈剂量依赖性。

Study on the Effects of Shenfu Yixin Decoction on Mitochondrial DNA Gene Expression in Primary Myocardial Cells under Hypoxia

To explore the effect of Shenfu Yixin Decoction on mitochondrial DNA gene expression in primary cardiomyocytes under hypoxic conditions.1~3 d SD newborn rats were taken, and primary cardiomyocytes were isolated, cultured and identified. CCK-8 was used to detect the optimal concentration of Shenxie Yixin and Coenzyme Q10. The experiment was divided into Normal group, Model (hypoxia) group, SFL (Shenyi Yixin side low dose) group, SFH (Shenyi Yixin side high dose) group, Coenzyme Q10 group, and the latter three groups in hypoxia. On the basis of the model, the cells were treated with Shenfu Yixin Recipe 0.25 mg/ml, Shenfu Yixin Recipe 0.5 mg/ml, Coenzyme Q10 1 × 10^-4 mol/L, and the cells were collected and tested for ATP content and lactic acid. Dehydrogenase (LDH) content, RT-PCR detection of ND-1, ND-2, COX, tfam gene expression. Under hypoxic conditions, compared with the normal group, the ATP of the myocardial cells in the model group was significantly decreased, the LDH was significantly increased, and the expressions of ND-1, ND-2, COX, and tfam genes were significantly decreased (P < 0.05, P < 0.01). Compared with the model group, the low dose group of Shenfu Yixin Decoction could reduce the LDH content and up-regulate the expression of ND-2 and COX genes, the difference was statistically significant (P < 0.05, P < 0.01). The high dose group of Shenfu Yixin Decoction could increase the ATP level of cardiomyocytes, decrease the content of LDH, and up-regulate the expression of ND-1, ND-2 and COX genes, the difference was statistically significant (P < 0.05, P < 0.01).Shenfu Yixin can up-regulate the expression of mitochondrial DNA-related genes COX, ND1 and ND2 in primary cardiomyocytes, increase myocardial ATP content and decrease LDH content in a dose-dependent manner.; increase myocardial ATP content; reduce LDH content, thereby protecting cardiomyocytes.