| ObjectiveWe observed the effects of Tongyangyiqi Method on angiogene sis by setting up myocardial infarction models, then do our res earch From the morphology(HE staining,CD34 staining),inflamm atory cytokines(IL-6),growth factors(TGF-β,VEGF,b FGF),signalin g pathway(MEK/ERK pathway). MethodsWe chose 80 healthy SD male rats,feeding in the environment of SPF.After one week,the model was copied with the ligation of left anterior descending of coronary artery(LAD).50 successful model rats were randomly divided into 5 groups(10 in each group):the Tongyangyiqi group(T),the Congbai group(C),the Huangqi group(H),the Shexiangbaoxinwan group(S),the Model group(M).At the same time,sham operated group(J) was also prepared.They were treated with different therapies for 4 weeks,then draw blood and clipping myocardial.Then we detected the expression of CD34 in the marginal zone of the infarction by immune histochemical method, studied the amount and density of capillaries in the models of every group.The pathological changes around the infarction area were observed by HE staining. Results1.Effects on myocardial ischemia angiogenesis tissue morphology of rats.High magnification showed that some myocardial cells bulged, interstitial cells swelled.And myocardial fiber dissolved or proliferated and inflammatory cell infiltration occurred in M group.Myocardial cell necrosis,inflammatory cell infiltration and fibrous proliferation were mild in C group and H group.While most of the myocardial cells kept normal and only a few of the myocardial cells necrosis, inflammatory cells infiltration occurred,and the fibrous proliferation was scanty in T group and S group.The myocardial cells and fibers kept normal in J group.The expression of CD34 in the marginal zone of the infarction by immune histochemical method showed that compared with the J group,MVD of the M group was significantly higher(p<0.01).MVD of all treatment groups were significantly higher than the M group(p<0.01).MVD of the T group had no significant difference from the S group.While the T group was significantly better than C group and H group(P<0.01).2.The content of serum IL-6 and TGF-β of rats with myocardial ischemia.Compared with the M group,IL-6 of the T group,C group and S group were significantly lower(p<0.01), which of the H group was also lower(p<0.05);IL-6 of the T group and C group were significantly lower than S group(p<0.05); while IL-6 of the T group had no significant difference from the C group(p>0.05).Compared with the M group,TGF- β of the C group was significantly lower(p<0.01),which of the T group and S group were also lower(p<0.05);while TGF-β of the T group had no significant difference from the C group and S group(p>0.05).3.The effects of VEGF and b FGF of rats with myocardial ischemia.Compared with the M group,VEGF/actin of all treatment groups were significantly higher(p<0.01),the expression of VEGF increased.The Amplification of VEGF m RNA was significantly higher(T group and S group were p<0.01,H group and C group were p<0.05);VEGF/actin of other treatment groups were lower than S group(p<0.01),the amplification of VEGF m RNA were also lower(p<0.01),the expression of VEGF reduced;Compared with the T group,VEGF/actin of both C group and H group were lower(C group was p<0.01,H group was p<0.05),the expression of VEGF reduced,the amplification of VEGF m RNA of H group was also lower(p<0.01),while there was no significant difference of the C group from the T group(p>0.05).Compared with the M group,b FGF/actin of T group and S group were significantly higher(p<0.01),the expression of b FGF increased,b FGF/actin of C group and H group had no significant difference(p>0.05),the amplification of all treatment groups were significantly higher(p<0.01).Compared with the S group,b FGF/actin of T group had no significant difference(p>0.05),but the amplification of b FGF m RNA was significantly lower(p<0.01);the amplification of b FGF m RNA of C group and H group were both lower than T group(C group was p<0.01,H group was p<0.05).4.The effects of p-MEK and p-ERK of rats with myocardial ischemia.Compared with the M group,p-MEK/actin of C group T group and S group were significantly higher(p<0.01),the expression of p-MEK increased,the amplification of the three groups were also significantly higher(p<0.01).Compared with the S group,p- MEK/actin of T group and C group had no significant difference(p>0.05),the amplification of p-MEK m RNA had no significant difference either(p>0.05) while which of C group was significantly lower(p<0.01).Compared with the T group,p-MEK/actin of H group was significantly lower(p<0.01),the expression of p-MEK reduced,which of C group had no significant difference(p>0.05),but the amplification of p-MEK m RNA of C group significantly reduced(p<0.01).Compared with the M group,p-ERK/actin of all treatment groups were significantly higher(p<0.01),the amplification of p-ERK m RNA were also significantly higher(p<0.01),the expression of p-ERK increased.Compared with the S group,p-ERK/actin of other treatment groups were significantly reduced(H group and C group were p<0.01,T group was p<0.05),the expression of p-ERK reduced,the amplification of p-MEK m RNA significantly reduced(H group and T group were p<0.01,C group was p<0.05);Compared with the T group, p-ERK/actin of H group was significantly lower(p<0.01),the expression of p-ERK reduced,but the amplification of p-MEK m RNA had no significant difference(p>0.05),there was no significant difference between the C group and the T group(p>0.05). ConclusionTongyangyiqi Method can increase MVC/MVD in the marginal zone of the myocardial infarction, and can promote the angiogenesis, the effect was better than H group or C group.Tongyangyiqi Method can reduce the level of IL-6,restrain inflammatory reaction,protect the endothelial cells which has been injuried,it can also increase the level of TGF-β.The effect of improving myocardial ischemia and promoting angiogenesis may be related to regulate the level of IL-6 and TGF-β.Tongyangyiqi Method can accelerate the expression of VEGF and b FGF,the effect was better than H group or C group.It means Tongyangyiqi Method has it’s advantage,the effect of promoting angiogenesis may be related to regulate the expression of VEGF and b FGF.Tongyangyiqi Method can accelerate the expression of p-MEK and p-ERK,the effect of promoting angiogenesis may be related to regulate the expression of beneficial growth factors and activate the MEK/ERK signal pathway.According to Tongyangyiqi Method,it has a good effect of promoting angiogenesis,which is better than Congbai group or Huangqi group,it can restrain inflammatory reaction, accelerate the proliferation of endothelial cells,which provided experimental basis for the clinical use of Tongyangyiqi Method.But the effct of some indicators is a little worse than Shexiangbaoxinwan.Maybe it was related to the error in experiments,after all the research of Shexiang baoxinwan has been quite mature.Our research still need further study. |
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