补阳还五汤对局灶性脑缺血大鼠海马神经可塑性蛋白表达的影响

目的采用多模态磁共振成像观察补阳还五汤对局灶性脑缺血大鼠海马组织结构、血流灌注的影响,结合神经可塑性蛋白及糖代谢相关转运蛋白表达变化分析其影响海马神经可塑性的作用机制。方法制备右侧大脑中动脉栓塞致局灶性脑缺血大鼠模型,将大鼠随机分为假手术组、假手术+补阳还五汤组、模型组、模型+补阳还五汤组,假手术组和模型组予生理盐水灌胃30 d,假手术+补阳还五汤组和模型+补阳还五汤组予补阳还五汤灌胃30 d。T2 mapping成像观察海马组织结构,动脉自旋标记成像检测海马血流灌注情况,Western blot检测海马组织神经可塑性相关蛋白突触素(SYN)、生长相关蛋白-43(GAP-43)、葡萄糖转运蛋白(GLUT)和单羧酸转运蛋白(MCT)表达。结果与假手术组比较,模型组大鼠右侧和左侧海马T2弛豫时间均显著增加(P<0.01,P<0.001),右侧海马血流量显著降低(P<0.05),右侧海马组织SYN、GAP-43、MCT4、MCT2蛋白表达显著降低(P<0.01,P<0.001),双侧海马组织GLUT1、GLUT3蛋白表达显著降低(P<0.05,P<0.01,P<0.001);与模型组比较,模型+补阳还五汤组大鼠右侧海马T2弛豫时间显著减少(P<0.05),右侧海马血流量显著升高(P<0.05),双侧海马组织SYN、GAP-43、GLUT1、GLUT3蛋白表达均显著升高(P<0.01,P<0.001),右侧海马组织MCT4、MCT2蛋白表达显著升高(P<0.01,P<0.001)。结论补阳还五汤减轻局灶性脑缺血大鼠海马损伤可能与改善其血流灌注、上调神经可塑性相关蛋白、促进海马轴突再生和突触重塑、调节神经细胞能量代谢有关。

Effect of Yiqihuoxue prescription on myocardial energy metabolism after myocardial infarction via cross talk of liver kinase B1-depen-dent Notch1 and adenosine 5'-monophosphate-activated protein kinase

OBJECTIVE: To investigate the effect of Yiqihuoxue prescription(YQHX) from Traditional Chinese Medicine(TCM) on myocardial glucose and lipid metabolism after myocardial infarction via the cross talk between the liver kinase B1(LKB1)-dependent Notch1 and adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK). YQHX was prepared with substances with properties that benefit, to activate blood circulation based on the TCM theory.METHODS: Animal models of myocardial infarction were established by ligating Sprague Dawley rats' left anterior descending coronary arteries. The animals were randomly divided into a myocardial infarction(MI) group, a YQHX group, a perindopril group, a γ-secretase inhibitor, Notch signal inhibitor(DAPT) group, a DAPT+YQHX group and a sham group. The related drugs were administered on the second day after operation, and changes in the relevant indexes were examined on weeks 1 and 4.Changes in cardiac structure and function were examined by echocardiography. The glucose and free fatty acids(FFA) were examined by ELISA. The expression of Notch, LKB1 and AMPK m RNA was examined by a real-time fluorescence quantitative method. The expression of glucose transporter 4(GLUT4), and the expression of total acetyl-Co A carboxylase(ACC) and its phosphorylation were examined by western blotting.RESULTS: Compared with the sham group, the expression of Notch, LKB1 and AMPK m RNA in the MI group was lower. Compared with the MI group, the expression of these m RNAs in the YQHX and perindopril groups was higher, and their expression in the DAPT group was lower. At all time points, the protein expression of GLUT4 and p ACC decreased in the MI group. On week 1, the expression of p ACC protein was higher. In the DAPT group, the expression of p ACC protein decreased. Compared with the YQHX group, the expression of p ACC protein in the DAPT + YQHX group was lower. On week 4,compared with the MI group, the expression of GLUT4 protein in the YQHX group and the perindo-pril group was higher. The expression of GLUT4 protein in the DAPT group decreased. Compared with the YQHX group, the expression of GLUT4 protein in the DAPT+YQHX group was lower. There was no significant difference in the expression of ACC protein between the groups.CONCLUSION: YQHX promoted cross talk between the LKB1-dependent Notch1 and AMPK in myocardial tissue after myocardial infarction. Furthermore,it regulated the glucose and lipid metabolism of cardiomyocytes at different time points, thereby ameliorating the cardiac energy metabolism via different mechanisms and protecting the heart.