| Objective: Prescription Yiqi Huatan Quyu (It conposed of gualou, xiebai, banxia, chuanxiong, chishao, sanqi, huangqi, guizhi medicine.)has efficacy of yiqiwenyang,huatanquyu,tongmaizhitong.And it has been proved significantly effective in treating coronary heart disease (CHD) through years of clinical observation.In order to futher the probe its action mechanism, the model of CHD rats was established by feeding high fat forage and injecting pituitrin in the abdominal cavity according to reference documents. At the same time,the animal was interfered with medicine.We observed it's effect on electrocardiogram ST-segment,the contents of blood-fat,ET,NO, 6-keto-PGF1α, TXB2.The change of PAI-1 and t-PA expression in the myocardial and aortic tissue were observed by immunohistochemistry method.Futhermore,we made observation with HE dyeing methods on effect of this formula on pathomorphology of myocardial and aortic tissue.Methods:Select male SD rats 60,weight from 160 to 180g.Rats drunk water and took food freely,and were raised in sanitary experiment lab that temperature was 18℃～22℃,bright and dim alternate every 12 hours.After 1 week,they were divided into 5 groups randomly,for each group 12 rats:Nomal control group,Model control group,Low-dose of Prescription Yiqi Huatan Quyu group, High-dose of Prescription Yiqi Huatan Quyu group,Compound danshen drop pill control group.Reference to relevant literature,in addition to the normal group, all groups were given high-fat forage for 6 weeks,at the end of 72,48,24hours before stomach-perfusion, the intraperitoneal injection of pituitrin (30u/kg) for 3 times,each time interval of 24 hours,the formation of coronary heart disease myocardial ischemia model.Rats except for normal control group are treated as well as set up models. Meanwhile control drug or treatment drug was given in each treatment group and the appropriate dose of distilled water was given to normal and model group. Rats in each group were given stomach-perfusion once a day and every time the drug volume was calculated by 1ml/100g, total dose for 6 weeks. At the six weekend of experiment,the rats were injected the pituitrin for the third time intraperitoneally after 24h,with 20% urethane (4ml/kg) intraperitoneal injection of anesthesia. Then the ECG ST segment changes were recorded by the connection of biological function of BL-420E+ experimental system.Then the rats were anesthetized and their blood were drawn out,the blood sample was made contrifugation with low temperature and the blood serum or blood plasma was isolated.The contents change of TC,TG,HDL-C,LDL-C, NO,ET, 6-keto-PGF1α,TXB2 were detected. Rat blood taken soon after the open-chest profile control to take the heart and thoracic aorta, in the apical take the right amount of myocardial tissue, the myocardial and aortic tissu with the pre-cooling was 0.9% Nacl injection flush, filter paper moisture, the into 4% paraformaldehyde solution in a fixed,regular production of tissue biopsies.The changes of PAI-1,t-PA expression and organization morphology in myocardial and aortic were detected by immunohistochemistry and HE staining.All the data were made statistics analysis with SPSS13.0 software.Result:1 Effect of Prescription Yiqi Huatan Quyu on electrocardiogram ST-segment of cornoary heart disease ratsThe electrocardiogram ST-segment of model group (0.28±0.04) were significantly higher than normal group (0.14±0.05) (P<0.01),which showed that CHD rats had obvious myocardial ischemia.After the intervention of medicine, the electrocardiogram ST-segment changes of each treatment group (0.19±0.03,0.20±0.04,0.20±0.05) were significantly lower than the model group (0.28±0.04) (P<0.01).There was no marked difference among the low-dose group,high-dose group and control group (P>0.05).2 Effect of Prescription Yiqi Huatan Quyu on contents of TC,TG, HDL-C,LDL-C in serum of cornoary heart disease rats The contents of TC,TG,LDL-C in serum of model group (3.30±0.42, 1.14±0.11,1.42±0.16) were significantly higher than normal group (1.68±0.21,0.53±0.06,0.86±0.08) (P<0.01),while the contents of HDL-C in serum of model group (0.44±0.10) were significantly lower than normal group (0.73±0.12) (P<0.01), which showed that CHD rats had obvious blood-fat metabolic disorder.After the intervention of medicine,the contents of TC,TG,LDL-C in serum of each treatment group were significantly reduced (P<0.01), while the contents of HDL-C in serum of each treatment group were significantly increased (P<0.01). Among them,high-dose group (1.77±0.14) and low-dose group (1.80±0.18) in reducing the role of serum TC levels than the control group (2.39±0.34) (P<0.01), while the high-and low-dose group between the was no statistical significance (P>0.05);high-dose group (0.57±0.08) in reducing the role of serum TG levels than low-dose group (0.84±0.11) and the control group (0.82±0.10) (P<0.01) , while the low-dose group and the control group,no statistical significance between the comparison (P>0.05);high-dose group (1.05±0.16,0.65±0.13),low-dose group (1.14±0.11,0.59±0.09) and the control group ( 1.11±0.16,0.62±0.09) between the reduction of serum LDL-C levels and increased HDL-C content in the role was no statistical significance (P>0.05).3 Effect of Prescription Yiqi Huatan Quyu on contents of NO, ET of cornoary heart disease ratsThe contents of NO in serum of model group (29.14±3.70) were significantly lower than normal group (44.02±4.49) (P<0.01), while the contents of ET in plasma of model group (91.06±8.44) were significantly higher than normal group (66.30±4.98) (P<0.01). After the intervention of medicine,the contents of NO in serum of each treatment group were significantly increased (P<0.01),while the contents of ET in plasma of each treatment group were significantly reduced(P<0.01). Among them, high-dose group (41.33±5.23,66.11±7.58), low-dose group (40.59±3.79,68.00±6.62) in the elevated serum NO levels and reduced the role of plasma ET levels were better than the control group (36.13±4.31,75.03±7.56) (P<0.05 or P<0.01), while the high-and low-dose was no statistical significance between groups (P>0.05).4 Effect of Prescription Yiqi Huatan Quyu on contents of 6-keto-PGF1α,TXB2 in plasma of cornoary heart disease ratsThe contents of 6-keto-PGF1αin plasma of model group (281.26±36.67) were significantly lower than normal group (411.96±45.81) (P<0.01),while the contents of TXB2 in plasma of model group (554.47±57.31) were significantly higher than normal group (294.17±48.92) (P<0.01).After the intervention of medicine,the contents of 6-keto-PGF1αin plasma of each treatment group were significantly increased (P<0.05 or P<0.01),while the contents of TXB2 in plasma of each treatment group were significantly reduced(P < 0.01).Among them,high-dose group (398.17±37.61,304.98±36.77), low-dose group (383.74±48.23,308.73±29.15) in the elevated plasma 6-Keto-PGF1αand reducing the role of plasma TXB2 levels were better than the control group ( 332.44±67.71,350.76±45.34) (P<0.05 or P<0.01),while the high-and low-dose was no statistical significance between groups (P>0.05).5 Effect of Prescription Yiqi Huatan Quyu on pathomorphology of myocardial and aortic tissue of cornoary heart disease ratsMyocardial tissue:normal rat myocardial cells arranged in neat rows cytoplasm uniform,elongated oval-shaped nucleus,located in the central myocardial fibers,myocardial fibers were bundle distribution,arranged in order, no inflammatory cell infiltration.Model group,cardiac hypertrophy, disarrangement,heterogeneous cytoplasm,nucleus swelling and deformation, there inflammatory cell infiltration, myocardial cell gap widened,connective tissue increased,myocardial fibers is incomplete,and part of the myocardial fiber atrophy or degeneration,vacuole-like changes in local shows. Myocardial tissue of rats treatment groups have different degrees of improvement.Among them,high-dose group than the neat arrangement of myocardial cells,the nucleus changes in smaller,more complete myocardial fiber structure,only see part of the gap widened between the myocardial cells;the control group and the low dose group there are different degrees of myocardial cell gap widened connective tissue and myocardial fiber atrophy.Aortic Organization: normal group, integrity of rat aortic endothelial cells, aortic intimal smooth and non-proliferation, no inflammatory cell infiltration. Model group,rat aortic intimal rough, thickened,with fibrous tissue hyperplasia,aortic endothelial cell swelling,inflammatory cell infiltration occurred.The treatment group showed steady improvements in aortic pathological damage,including high-dose group,only see the aortic intimal rough; the control group and the low dose group visible aortic intimal rough, thick, mild proliferation of fibrous tissue.6 Effect of Prescription Yiqi Huatan Quyu on PAI-1,t-PA expression of myocardial and aortic tissue of cornoary heart disease ratsModel group myocardium and aortic PAI-1 expression (0.269±0.023, 0.251±0.026) compared with the normal group (0.121±0.027,0.126±0.024) increased (P<0.01), t-PA expression (0.096±0.021,0.150±0.039) compared with the normal group (0.219±0.040,0.283±0.036) decreased (P<0.01).After drug intervention,the drug group myocardium and aortic expression of PAI-1 decreased (P<0.01), t-PA expression increased (P<0.01).Among them, high-dose group (0.125±0.024,0.145±0.020,0.205±0.026,0.261±0.038), low-dose group (0.133±0.022,0.159±0.028 ,0.196±0.028,0.255±0.046) in the weakened heart and aorta PAI-1 expression and increased myocardial and aortic t-PA expression in the role of the superior to the control group (0.166±0.023,0.184±0.026,0.167±0.024,0.211±0.023) (P<0.05 or P <0.01),The high-and low-dose group had no statistical significance between the comparison (P> 0.05).Conclusions:1 Prescription Yiqi Huatan Quyu could correct the change of ECG ST segment ,reduce the contents of TC,TG,LDL-C,increase the contents of HDL-C of cornoary heart disease rats.It had marked function in improving myocardial ischemia and regulating blood-fat.2 Prescription Yiqi Huatan Quyu could significant improve the myocardial and aortic tissue lesion degree,it manifesed like this:after the drug was given, myocardial cells arranged in neat rows, nuclei and myocardial fiber structure of basic integrity, vacuole-like change and aortic wall membrane proliferation significantly reduced ,and inflammatory cell infiltration almost disappeared.It indicated that Prescription Yiqi Huatan Quyu had protective effect obviously to myocardial and aortic tissue.3 Prescription Yiqi Huatan Quyu could significant elevat serum NO and plasma 6-Keto-PGF1αcontent,reduc levels of plasma ET and TXB2,balance NO, ET and 6-Keto-PGF1α,TXB2 secretion and release,with Protection of vascular endothelial function.4 Prescription Yiqi Huatan Quyu could inhibit the PAI-1 expression of myocardial and aortic tissue , increase myocardial and aortic t-PA expression , restore the equilibrium of fibrinolytic system,prevent atherosclerosis and thrombosis, in order to achieve The purpose of treatment of CHD. |
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