活血潜阳方对“血瘀-阳亢-痰浊”高血压模型大鼠胰岛素信号通路基因表达及心肌重构的影响

Effects of Huoxue Qiangyang Fang on Insulin Signaling Pathway Gene Expression and Myocardial Remodeling in Xueyu-Yangkang-Tanzhuo Syndrome Model of Spontaneous Hypertensive Rat

目的:探讨活血潜阳方对"血瘀-阳亢-痰浊"高血压模型大鼠胰岛素信号通路基因表达及心肌重构的影响。方法:以36只5周龄雄性自发性高血压大鼠(spontaneous hypertensive rat,SHR)为对象,按照随机数字表法按2∶1随机分为两组:SHR空白对照组(SHR-C组)、SHR模型组;SHR模型组通过附子汤灌胃+高脂饮食4周后随机分为SHR模型对照组(SHR-M组)及SHR模型治疗组(SHR-H组),9只同龄京都大鼠(WistarKyoto,WKY)为正常对照组(WKY-C组),9周龄时处死并取其心脏进行组织切片,提取心肌组织RNA用胰岛素功能基因芯片筛选效应基因,从中选择部分主要的关键基因,用荧光定量PCR法检测葡萄糖激酶(GCK)mRNA、6-磷酸葡萄糖酶催化亚基(G6PC)mRNA及丙酮酸脱氢酶激酶4(Pdk4)mRNA水平,Western Blot检测GCK、G6PC及Pdk4蛋白表达,在光镜及电镜下大鼠心肌结构。结果:SHR-H组大鼠心肌GCK、G6PC表达明显上调(P<0.01),Pdk4表达明显下调(P<0.05),光镜及电镜下观察发现SHR-H组大鼠心肌结构较SHR-M组明显改善。结论:活血潜阳方具有改善SHR模型大鼠心肌重构的作用,其机制可能与上调心肌组织GCK、G6PC蛋白表达、下调心肌组织Pdk4蛋白表达有关。

Objective：To discuss the effect of Huoxue Qianyang Formula （HXQYF） on insulin - signaling pathway gene -expression and myocardial remodeling in Xueyu Yangkang Tanzhuo Syndrome model of spontaneous hypertensive rat model. Methods：36 male SHRs, five -week - old for the object, according to the random number table 2 ： 1, were ran- domized into two groups： SHR control （SHR -C） group, SHR model group,which were fed with monkshood and high fat diet at the first four weeks, which were randomly divided into SHR - M group and SHR model treatment （ SHR - H） group,9 sex and age matched Wistar- Kyoto rats（WKY） were served as another control group（WKY- C ）. They were sacrificed and their hearts were tissue sections in 9 weeks old, extracted RNA with insulin function in myocardial tissue microarray gene - screening - effect, choosing the part of key genes. Realtime fluorescence quantitative PCR （ RT - PCR） was applied for the detection of expression levels of subunit mRNA of the enzyme glucokinase （GCK） , glucose -6 -phos- phate （G6PC） and pyruvate dehydrogenase kinase 4 （Pdk4）. Western Blot was applied to detect its protein expression, u- sing the light and electron microscopic to observe the myocardial structure. Results ： Compared with SHR - M group, the ex- pression of GCK and G6PC of SHR - H group was significantly up - regulated and the expression of Pdk4 was significantly reduced. Using light microscopy and electron microscopy, we found that the myocardial structure was significantly im- proved in SHR - H group compared with SHR - M group. Conclusion ： HXQYF can improve myocardial remodeling of SHR rat model,which may be related by increasing the myocardial remodeling protein expression of GCK and G6PC and reduc- ing the protein expression of Pdk4.