| Coronary heart disease can be described as "obstruction", "heartache" in TCM. Acute myocardial infarction (AMI) belongs to the severe symptom of "obstruction" or "heartache". The basic mechanism of "obstruction" and "heartache" is deficiency of "BEN" and sufficiency of "SHI". The deficiency of "BEN" means the shortage function of heart, kidney, QI and blood. The sufficiency of "SHI" means blood stasis, QI stagnant and cold stagnation. Controlling left ventricular remodeling after AMI is fundamental to prevent the occurrence of heart failure and improving the prognosis of AMI patients. There are no specific anti-ventricular remodeling drugs in home and abroad. HuoXueAnXinFang is constituted by effective components of Chinese herbs which have specific targets and were prescribed according to the treatment principles:benefiting vital energy and activating blood circulation, nourishing heart and calm the nerves, eliminating sputum and removing stasis. The clinical curative effects of HuoXueAnXinFang is distinct. One object of this study is to examine the anti-ventricular remodeling effects of five different compatibility of HuoXueAnXinFang on Hyperlipidemia AMI rats and chose the most effective one. The other object is to identify the anti-ventricular remodeling mechanism of the optimized compatibility of HuoXueAnXinFang.Object ive1 The first aim was to study the anti-ventricular remodeling and hyperlipidemia regulating effects of HuoXueAnXinFang. HuoXueAnXinFang was divided to five different effective components groups according to the monarch, minister, assistant and guide principle and was treated to hyperlipidemia AMI rats for 7d and 21d respectively.2 The other object was to further study the anti-ventricular remodeling mechanism of the most effective components group of HuoXueAnXinFang selected by pharmacological study. We studied the mechanism from its anti-inflammatory, anti-oxidative damage, matrix metalloproteinases inhibiting and angiogenesis promotion.Methods1 pharmacological part Wistar rats were fed with high fat food for 15d, then the hyperlipidemia rats were induced to hyperlipidemia AMI rats by occluding the left descending coronary artery. HuoXueAnXinFang effective components treated to hyperlipidemia AMI rats for 7d or 21d respectively from the day of surgery. The rats were divided to 9 groups: normal, sham, hyperlipidemia AMI model, positive Chinese medicine control and five different effective components groups of HuoXueAnXinFang. Each group has 15 rats. The drugs were gavaged to rats and the same volume of drinking water also gavaged to normal, sham and the AMI model rats. The five different effective components of HuoXueAnXinFang groups are the following:group one, the monarch part of salvia miltiorrhiza total phenolic acids; group two, monarch part of salvia miltiorrhiza total phenolic acids, minister part of panax notoginseng and assistant part of total nucleoside adjuvant zi extracts; group three,monarch part of salvia miltiorrhiza total phenolic acids, minister part of panax notoginseng and minister part of Melon trichosanthis and allium macrostemon extracts; group four, monarch part of salvia miltiorrhiza total phenolic acids, assistant part of ginkgo biloba extract and assistant part of total nucleoside adjuvant zi extracts and guide part of astragalus extract; group five, monarch part of salvia miltiorrhiza total phenolic acids, minister part of panax notoginseng and minister part of Melon trichosanthis, assistant part of ginkgo biloba extract and assistant part of total nucleoside adjuvant zi extracts and guide part of astragalus extract. Index related with ventricular remodeling were examined including cardiac index, myocardial infarction area, noninvasive heart function testing of ventricular inner diameter and ventricular capacity, and left ventricular ejection fraction fractional shortening and myocardial morphological structure study by paraffin imbed and H & E dyeing and electron microscope. The anti-ventricular remodeling effects of HuoXueAnXinFang in different components groups were thoroughly examined.2 The anti-ventricular re modeling mechanism part The hyperlipidemia AMI rats model were established as described in part 1, HuoXueAnXinFang treated for 7d and 21 d respectively. Each treating time were 4 groups: sham control group, the hyperlipidemia AMI model group, the positive control of compound danshen drop pill and effective components of HuoXueAnXinFang group. RT-PCR method was used to detect left ventricular myocardial infarction area, the border area and the noninfarction area of TLR4 and NFκB mRNA expression, WB method was used to detect left ventricular myocardial infarction area, the border area and the noninfarction area of TLR4 and NFκB protein expression, ELISA test was used to detect the left ventricular myocardial infarction area, the border area and the noninfarction area of TNF alpha and IL1βlevel in serum. discusses the effective components at party blood compatibility of inflammatory damage in the whole process of ventricular remodeling the role of mechanism; UV spectrophotometer method was used to detect left ventricular myocardial infarction area, the border area and the noninfarction area of iNOS and eNOS vitality, endogenous antioxidant enzymes tSOD vitality and the peroxide product content of MDA. The RT-PCR method was used to detect left ventricular myocardial infarction area, the border area and the noninfarction area of HIF1 alpha and VEGF mRNA expression, WB method was used to detect left ventricular myocardial infarction area, the border area and the noninfarction area of HIF1 alpha and VEGF protein expression, ELISA test was used to detect left ventricular myocardial infarction area, the border area and the noninfarction area of matrix metalloproteinases MMP9 and TXB2 levels in serum.Results1 The ventricular remodeling progress of hyperlipidemia AMI rats 7d after occluding the left descending coronary the left ventricular pathological changes were:heart index and the left heart index increased significantly, left ventricular end-diastolic and end-systolic diameter and capacity increased significantly, heart ejection fraction and left fractional shortening decreased significantly; 21 d after occluding the left descending coronary the left ventricular pathological changes were:the whole heart index and left heart index was significantly higher than the sham group but without further increase compared to 7d group, myocardial infarction area is further expanded compared to 7d group (56.43±11.32 VS 44.26±8.18) and myocardial collagen deposition and fibrosis is obviously severer than 7d group, left ventricular diastolic inside diameter and left ventricular systolic inside diameter further significantly increased than 7d group, especially left ventricular end-diastolic diameter expanding more obvious (LVIDd:0.71±0.06 VS 0.64±0.05, LVIDs:0.50±0.05 VS 0.48±0.05), left ventricular end-dias tol ic capacity and end-systolic capacity is further increased significantly than 7d group(EDV:0.78±0.14 VS 0.50±0.06, ESV:0.42±0.12 VS 0.17±0.05), stroke volume lower than 7d group (SV:0.34±0.12 VS 0.30±0.08).2 Five effective components of HuoXueAnXinFang on hyperlipemia AMI rats about anti-ventricular remodeling effects Treating for 7d:group three and five of HuoXueAnXinFang effective decreased the infarction area, narrowed left ventricular end-diastolic inside diameter and left ventricular end-systolic diameter and had advantage compared to group one. Group five treating for 7d end-systolic capacity had significantly decreased and heart left ventricular ejection fraction and fractional shortening significantly increased. Group three and group five treating for 7d can see profile of outside, muscularis and inner layer and some remaining myocardial cells in infarction area, collagen deposition and inflammatory cells infiltrating significantly reduced compared to model group; Treating for 21d:the group five of effective components of HuoXueAnXinFang reduced the whole heart index and left heart index significantly, decreased left ventricular infarction area, decreased left ventricular end-diastolic inside diameter and left ventricular end-systolic diameter, reduced left ventricular end-diastolic volume and end-systolic volume, elevated left ventricular ejection fraction and fractional shortening.3 five effective components of HuoXueAnXinFang on hyperlipemia AMI rats about adjusting the blood lipid Treating for 7d:the five groups all effectively reduced serum CHO, TG and VLDL level. Group two can reduce serum LDL level. Treating for 21d:group four and group five reduced all indexes of blood lipid in serum effectively. Group three reduced TG, LDL and VLDL effectively. Group four and Group five had advantage of regulating blood lipid than Group one and Group two.4 The molecular mechanism of ventricular remodeling on hyperlipidemia AMI rats Inflammatory damage:7d after surgery, left ventricular myocardial infarction area and border area TLR4 and NFκB mRNA and protein expression significantly increased, inflammatory cytokine TNF alpha and ILlbeta increased significantly in serum, myocardial infarction area and border area.21d after surgery,left ventricular myocardial border area and the noninfarction area TLR4 and NFκB mRNA and protein expression significantly increased, inflammatory cytokine TNF alpha and ILlbeta significantly increased in myocardial border area and the noninfarct ion area and serum TNFa also increased significantly; The peroxidation damage:7d after surgery, the infarction area, border area and the noninfarct ion area the vitality of the i NOS were significantly increased, and in the same area of the vitality of eNOS decreased significantly, the antioxidant enzymes tSOD vitality decreased significantly in left ventricular myocardial infarction area and border area, peroxide product MDA significantly increased in myocardial infarction area, border area, the noninfarction area and in the serum.21d after surgery, the vitality of the iNOS were significantly increased in myocardial border area and the noninfarction area, Serum iNOS and eNOS vitality changes were not apparent, the antioxidant enzymes tSOD vitality decreased significantly in left ventricular myocardial infarction area, border area and noninfarction area, peroxide product MDA significantly increased in myocardial infarction area, the noninfarct ion area and in the serum. Angiogenesis:left ventricular myocardial infarction area HIF-1 alpha and VEGF mRNA and protein expression levels are increased 7d and 21d after surgery, the border area and the noninfarct ion area of HIF-1 alpha and VEGF mRNA and protein expression level did not rise. MMP9 protein level increased obviously in myocardial infarction area and border area 7d and 21d after surgery. The TXB2 level did not show any difference among groups.5 The anti-ventricular remodeling mechanism of HuoXueAnXinFang Effective components Anti-inflammatory damage mechanism: HuoXueAnXinFang treating for 7d significantly decreased TLR4 and NFκB mRNA and protein expression levels in myocardial infarction and border area, significantly decreased TNF alpha and IL1βexpression level in myocardial infarction area and border area. HuoXueAnXinFang treat ing for 21d significantly decreased TLR4 and NFκB mRNA and protein expression level in myocardial border area and the noninfarction area, significantly decreased the TNF alpha and IL1βexpression level in myocardial infarction area and the border area, TNF alpha level in serum also decreased. HuoXueAnXinFang treating for 21d had advantage on TLR4 and NFκB mRNA and protein regulating compared to positive control group; The anti-peroxidation damage mechanism:HuoXueAnXinFang treating for 7d significantly decreased iNOS vitality in myocardial infarction area and border area, raised eNOS vitality in infarction area, border area and the noninfarction area, significantly reduced serum and myocardial regional of MDA level, increased the vitality of tSOD in myocardial border area; HuoXueAnXinFang treating for 21d significantly decreased iNOS vitality in myocardial border area and noninfarction area, increased eNOS vitality in myocardial infarction area, border area and noninfarction area, significantly decreased MDA content in myocardial border area, noninfarction area and serum, significantly increased the tSOD vitality in myocardial border area and noninfarction area. HuoXueAnXinFang treating for 21 d had advantage on iNOS and tSOD vitality regulating compared to positive control group; promoting Angiogenesis mechanism:substrate biodegradation and blood clots state: HuoXueAnXinFang treating for both 7d and 21d increased the HIF-1 alpha and VEGF mRNA and protein expression level in left ventricular myocardial infarction area compared to model or positive control group, significantly reduced MMP9 level in left ventricular myocardial infarction area and border area. Both HuoXueAnXinFang and positive control medicine had no regulating effects on TXB2 level.Conclus ion1 From 7d to 21d after surgery the ventricular remodeling progress increased gradually on hyperlipidemia AMI rats. The five group of HuoXueAnXinFang effective components had the best anti-ventricular remodeling and regulating blood lipid effects among the five different groups. The effects had advantages compared to positive control Chinese med ic ine.2 Inflammatory damage plays an important role in the whole process of ventricular remodeling, oxidative stress induced apoptos is promoted the developing of further development of ventricular remodeling, the degradation of ECM and angiogenesis play roles in the worsen stage of ventricular remodeling. HuoXueAnXinFang effective components had effects of anti-inflammatory, antioxidant and promote angiogenesis, its anti-inflammatory and antioxidant effect is better than the positive control Chinese medicine.3 The protection effects of HuoXueAnXinFang effective components on ventricular remodeling were fulfilled by its anti-inflammatory, antioxidant, inhibiting ECM degradation and promoting angiogenesis.
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