Effect Of Luo Fengning No. 1 On Coronary Vascular Endothelial Function In Rats With Myocardial Infarction. Chinese Journal Of Integrative Medicine On Cardio / Cerebrovascular Disease]

BackgroundCoronary heart disease(CHD), which severely endanger human health, is a type of cardiovascular disease with high incidence. Traditional Chinese Medicine (TCM) stress syndrome differentiation and overall adjustment the function of the body, which has a unique advantage in the treatment of CHD. According to the characteristics of CHD, professor X put forward the theory that:endogenous collateral wind is another important pathogenesis of CHD, and create a herbal compound which is called Luofengning-1 for the treatment of CHD.ObjectivesAccording to systematic review, we investigate the clinical effects of calm the wind and activating collaterals herbal compound on CHD.Using the methods of molecular biology to detect the content of related molecular of coronary vascular endothelial injury, we explore the mechanism of Luofengning-1 formula on CHD, and investigate the details in regulating CHD.MethodsWe used the RevMan 5.1.0 statistical software to analysis of data of theraperutic effect calm the wind and activating collaterals drugs on CHD.We establish myocardial infarction rat model by ligaturing the left anterior desceding coronary artery. According to ECG evaluation all successfully modeled rats were randomly divided into the sham-operation group, the model group, Luofengning-1 group, drug combination group, captopril group. The rats in each group were given corresponding drugs by gavage twice a day after surgery, they all treated for 4 weeks. After 4 weeks, we observe the changes of experimental lesions of myocardial infarction and related indexes of cardiac function in rats to evaluate the success rate of myocardial infarction model making and efficacy of Luofengning-1 on treatment of rats with myocardial infarction. To detect the changes of mRNA expression and protein of EPCR, TM of myocardial in infarct border zone, serum sEPCR and sTM content, plasma NO and PGI2 content, we explore the regulatory mechanism of Luofengning-1 formula on anticoagulation substances of vascular endothelium; To detect the changes of NF-kB, VCAM-1 and ICAM-1 expression of myocardial in infarct border zone, we explore the regulatory mechanism of Luofengning-1 formula on anti-inflammatory substances of vascular injury.ResultsThe systematic review identified 7 eligible trials, and contained 478 participants, the methodological quality of all the trials was low. The results of this systematic review shows the efficacy and safety of calm the wind and activating collaterals herbal compound on the treatment of CHD. Calm the wind and activating collaterals herbal compound showed a statistically significant effect on angina relief (RR=1.31,95%CI=1.06-1.64), and improvement in electrocardiogram(ECG) (RR=1.57,95%CI=1.18-2.11) compared with compound salvia pellet, one RCT showed that calm the wind and activating collaterals herbal compound can significantly reduced TXB2/6-PGF1α(RR=-0.89,95%CI=-1.25--0.53), but there were no statistically significant differences between experimental and control groups in TCM syndromes, blood rheology, blood lipid, C-reactive protein. Calm the wind and activating collaterals herbal compound were found to markedly improved angina relief(RR=1.44, 95%CI=1.04-2.00), ECG(RR=1.53,95%CI=1.02-2.31), and TCM syndromes(RR=1.44, 95%CI=1.04-2.00) compared with ginkgo leaf tablet, but there were no statistically significant differences between experimental and control groups in blood lipid. Three RCT reported calm the wind and activating collaterals herbal compound plus routine western medicine compared with routine western medicine, and two trials showed significant effect on angina relief (RR=1.31,95%CI= 1.07-1.60), two trials were found to markedly improved ECG (RR=1.44, 95%CI=1.16-1.79), but there were no statistically significant differences between experimental and control groups in blood lipid, inflammatory factors, calm the wind and activating collaterals herbal compound plus routine western medicine were found to markedly reduced high blood viscosity(MD=0.27,95%CI=0.05-0.49) compared with Tongxinluo capsule plus routine western medicine, but there were no statistically significant differences between experimental and control groups in angina relief, ECG, TCM syndromes. Adverse effects were mentioned in 3 trials, one studies reported no adverse effects, the remaining two trials reported adverse events including gastric discomfort, diarrhea, there were no statistically significant differences between experimental and control groups in these studies.The experimental research consists of three parts.In experimental study 1, we successfully established rats myocardial infarction model. Compared with preoperative ECG, postoperation rats in sham group had no change, while postoperation rats in myocardial farction model had significant changes:ECG had persistent ST-segment elevation and Q waves in I, aVL, Ⅴ1-Ⅴ6, but there was no statistically differences in myocardial farction model group. Luofengning-1 group can improved the TCM symptom of heart-blood stasis, and Luofengning-1 and Captopril showed good synergistic interaction in improving the TCM symptom of heart-blood stasis. Compared with sham group, the echocardiogram parameters including LVPWTd, LVPWTs, EF%, FS% in model group had obvious decrease, and LVDs, LVDd, organ body weight ratio had increase. Compared with model group, LVPWTd, LVPWTs, EF%, FS%, LVDd in Luofengning-1 group had obvious increase but all these has no statistically significant difference, and LVDs, organ body weight ratio had decrease, and organ body weight ratio has statistically significant difference(P< 0.05)Compared with Captopril group, LVDd, LVDs, organ body weight ratio in Luofengning-1 group had increase slightly, and LVPWTd, LVPWTs EF%, FS%, had decrease, except for organ body weight ratio has statistically significant difference(P<0.05), the others has no statistically significant difference. There was no difference between Luofengning-1 group and model group in pathological changes, but the lesion range in Luofengning-1 group had markedly reduce compared with model group, the lesion range in Luofengning-1 group had increase compared with Captopril group. Compared with model group, LVPWTd, LVPWTs, EF%, FS% in drug combination group had obvious increase, and LVDs, LVDd, organ body weight ratio had decrease, among them LVPWTs has statistically significant difference(P< 0.05), organ body weight ratio has markedly statistically significant difference(P<0.01). Compared with Captopril group. LVDs, LVDd LVPWTd, LVPWTs in drug combination group had increase, and EF%, FS% had decrease, but all these has no statistically significant difference. There was no difference between drug combination group and model group in pathological changes, but the lesion range in drug combination group had markedly reduce compared with model group, and had reduce compared with Captopril group.In experimental study 2, we detect the mRNA and protein of TM and EPCR expression in the myocardial infarct border zone measured by real-time PCR and western blot respectively, and test the serum sEPCR, sTM content measured by enzyme-linked immunosorbent assay(Elisa). Compared with the sham group, the experimental results showed that the mRNA expression of EPCR and TM in the model group increased significantly (P<0.01), protein content expression of EPCR and TM decreased obviously(P <0.01, P<0.05), the serum levels of sEPCR and sTM increased significantly(P<0.01). Compared with the model group, the results showed that the mRNA expression of EPCR and TM in the Luofengning-1 group decreased significantly (P<0.01), protein content expression of EPCR obviously increased (P<0.01), protein content expression of TM increased(P> 0.05), the serum levels of sEPCR and sTM decreased (P<0.01,P<0.05). Compared with the Captopril group, the results showed that the mRNA expression of EPCR and TM in the Luofengning-1 group increased significantly (P<0.01), protein content expression of EPCR and TM decreased (P>0.05), the serum levels of sEPCR and sTM increased obviously (P< 0.01). Compared with the model group, the results showed that the mRNA expression of EPCR and TM in the drug combination group decreased significantly (P<0.01), protein content expression of EPCR and TM obviously increased (P<0.01,P>0.05), the serum levels of sEPCR and sTM decreased (P<0.01). Compared with the Captopril group, the results showed that the mRNA expression of EPCR and TM in the drug combination group decreased(P>0.05), protein content expression of EPCR and TM increased (P>0.05), the serum levels of sEPCR and sTM decreased (P>0.05). The experimental results showed that the plasma level of NO and PGI2 in the model group decreased significantly compared with the sham group (P<0.01). The plasma level of NO and PGI2 in the Luofengning-1 group increased compared with the moded group (P>0.05), and also increased compared with the Captopril group (P>0.05). The plasma level of NO and PGI2 in the drug combination group increased significantly compared with the sham group (P<0.01), and also increased compared with the Captopril group (P>0.05).In experimental study 3, we detect the positive expression of NF-kB and integral optical density of VCAM-1 and ICAM-lby immunohistochemical method. The result showed that the positive expression of NF-kB and integral optical density of VCAM-1 increased compared with sham group(P<0.01). The positive expression of NF-kB and integral optical density of VCAM-1 in Luofengning-1 group decreased compared with model group(P<0.01,P<0.05), and integral optical density of ICAM-1 in Luofengning-1 group decreased compared with model group(P>0.05). The positive expression of NF-kB and integral optical density of VCAM-1 and ICAM-1 in Luofengning-1 group increased compared with Captopril group(P >0.05). The positive expression of NF-kB and integral optical density of VCAM-1 in drug combination group decreased compared with model group(P<0.01), and integral optical density of ICAM-1 in drug combination group decreased compared with model group(P> 0.05). The positive expression of NF-kB and integral optical density of VCAM-1 and ICAM-1 in drug combination group decreased compared with Captopril group(P>0.05).ConclusionCalm the wind and activating collaterals herbal compound may have beneficial effects in improving angina relief, ECG, and TCM syndromes, but the low quality of existing trials makes it difficult to draw a satisfactory conclusion.Luofengning-1 herbal compound can improve the TCM symptom in rats, Luofengning-1 herbal compound played a minor role in improving the cardiac structure and function, slightly reduced infarct size, and can not show good synergistic interaction in improving the cardiac function and reducing infarct size with Captopril in this experimental study.Luofengning-1 herbal compound can improve the anticoagulant effect in endothelial cells, reduce injuries of EPCR and TM, inhibit compensatory increase of mRNA expression of EPCR and TM, increase the protein expression of EPCR and TM, decreased plasma content of sEPCR and sTM, and Luofengning-1 herbal compound and Captopril showed good synergistic interaction.Luofengning-1 herbal compound can reduced NF-kB、VCAM-1 expression in the myocardial infarct border zone, and Luofengning-1 herbal compound and Captopril show good synergistic interaction, but Luofengning-1 herbal compound can slightly reduced ICAM-1 expression in the myocardial infarct border zone.