| Objective To observe the treatment of Xin-Yi capsule on myocardial ischemia-reperfusion injured rats,and to explore the correlation of Rho/Rho kinase signaling pathway and myocardial ischemia reperfusion injury.Methods SD rats were randomized into six groups, such as the model group(the same volume of distilled water), the Xin-Yi Capsule high, medium and low dose groups(at the daily dose of 8.4 g /kg, 4.2 g /kg and 2.1 g /kg), the control groups(compound danshen dripping pills,at the daily dose of 30 mg/kg), sham group(the same volume of distilled water), Successive administration for 7 days.After the last administration, the rats were treated by closing the left anterior descending coronary artery for 30 min and then releasing it for 120 min under anesthesia.S-T segment change was recorded,Blood samples were collected from the abdominal vein of the animals, and the serum was collected for the measurement of CK-MB、LDH、MCP-1、IL-6、SOD、MDA by ELISA, myocardium tissue morphology was measured by HE staining. The Rho A,MLCP expression by immunohistochemical method in the myocardial tissue, Blodsampewrctdfomheabinlvoftheanimls,deruwascoltdfrhe The apoptosis of cells in myocardium tissue was measured by(terminal dexynucleotidyl transferase(Td T)-mediated d UTP nick end labeling)TUNEL staining,the Rho kinase level were measurd by RT-PCR and MYPT-1were measurd by Western-Blot.Results1.The magnitude of ST segment Compared with sham group, S-T segment elevation was elevated obviously in(myocardial ischemia) 0.5 min,(reperfusion) 0.5 min, 1 min, 2 min in Model group rats(p<0.01);Compared with model group, the experiment revealed that Xin-Yi Capsule attenuated S-T segment elevation(p<0.05, p<0.01).2.Serum biochemical indexes Compared with sham group, the levels of serum CK-MB、LDH、MCP-1、IL-6、MDA were significantly higher(P<0.01) and SOD was decreased in model group;Compared with model group, the the levels of serum CK-MB、LDH、MCP-1、IL-6、MDA decreased and SOD increased significantly in drug groups(p<0.05, p<0.01).nclear,iftonmaycels3.HE staining results From pathology testing,The sham group showed myocardial fiber integrity, cells arranged closely, nucleolus was clear, a clear cross striations, no necrosis and inflammatory cell infiltration. In the model group rats myocardial fiber fracture,myocardial cells arranged in disorder, nuclear pyknosis, cell morphological ambiguity,stripes is unclear, infiltration of inflammatory cells, intercellular space enlarged etc.Positive group, Xinyi capsule high dose group, middle dose group and low dose group compare with model group inflammatory cell infiltration was light, and myocardial cells arranged in neat rows, myocardial trend is clear.Inthemodlgroupatsmyocardilfiberacture,myocardilesarngedindisore,nuclarpyknosi,celmorphlgicambiguty,stripesunclear,infltraofinlamtorycels The results of Rho/Rho kinase signaling pathway related protein.4. The expression of related protein in Rho/Rho kinase signal pathway Immunohistochemical staining result,Compared with sham group,myocardial tissue Rho A and MLCP levels were higher significantly(p<0.01);Inthemodlgrupatsmyocrdilfberactu,myodilcesarngdisorde,nuclapyknosi,celmorphgicambuty,sripesunclear,iftoninflamtoryces Compared wihe model group,the levels of Rho A and MLCP dropped in drug groups(p<0.05).xpresionf Rhkadug The expression levels of Rho kinase m RNA in each group were detected by the ratio of Rho kinase to β-actin light density. Inthemodlgrupatsmyocrdilfbeatu,myocrdilesangidsore,nuclapyksielmorphgicabuty,sripeuncla,iftronlamtoryces,intrluapcenlargdt Compared with sham group,Rho kinase m RNA increased significantly Compared wihe model group,the expression of Rho kinase dropped in drug groups(p<0.01).Compared with sham group,The expression of p MYPT1 protein was increased in Model group by WB(P<0.01); Compared with model group, The expression of p MYPT1 protein was decreased in drug groups(p<0.05, p<0.01)).5.Apoptosis of cardiac muscle cells by TUNEL Compared with sham group,the model group increased the rates of cardiocytest apoptosis(p<0.01);Compared with model group,the apoptosis was decreased in each treatment group.Conclusion Xin-Yi Capsule have a protection function for myocardial ischemia reperfusion iniury,its mechanisms may be ralated to the Rho/Rho kinase signal pathway. |
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