The Pirmary Discussion Of The Effect And Mechanism On Rutin Reserving Myocardial Fibrosis In Rats

Background: Myocardial fibrosis is not only a result caused by all kinds of heartdiseases, but also a precipitating factor for these diseases．Avoiding the occurrence ofmyocardial fibrosis or delaying its development has a profound meaning for a variety ofcardiovascular disease. So we need to know the mechanism of the occurrence anddevelopment of myocardial fibrosis exactly. Rutin can protect cardiomyocyte, so theexperiment had a preliminary discussion about the effect and mechanism of Rutin onrebersing myocardial fibrosis.Objective: To discusses the mechanism of myocardial fibrosis as well as theintervention effect of Rutin on myocardial fibrosis by analyzing the expression of ColⅠ,Col Ⅲ, TGF-β1, CTGF, PI3K and Akt, p-Akt in normal and abnormal myocardial tissueof rats.Methods:1. In vivo experiment, myocardial fibroblasts induced by Ang II,6dosegroups were divided, including C group, M group, Val group(10-6mg·L-1), R groups (1,5,10mg·L-1). The proliferation of cardiac fibroblast and the expression of ColⅠ, Col Ⅲ,TGF-β1, CTGF, PI3K and Akt were detected through MTT, Elisa, RT-PCR and WesternBlotting. At the same time, the correlations were analyzed.2. In vivo,48male Wistar rats were selected and divided into6groups randomly,including C group, M group, Val group (10mg·kg-1·d-1), R groups(10,25,50mg·kg-1·d-1).The M group was established by multipoint injecting Iso subcutaneously. The rats weregiven a dose each day through gavaging.3weeks later, the samples of rats’ hearts wereexamined by HE staining, and detected the expression of ColⅠ, Col Ⅲ, TGF-β1, CTGF,PI3K and Akt, p-Akt via immunohistochemistry, RT-PCR, and Western Blotting. At thesame time, the correlations were analyzed.Result:1. In vivo experiment:①The result of MTT, in the AngII induced cardiacfibroblast after treating24h, the over proliferation of cardiac fibroblast was significant(P<0.01). The proliferation of cardiac fibroblast could be inhibited by different density ofRutin significantly (P<0.01).②The result of Elisa, in the Ang II-induced cardiac fibroblast after treating24h, the content of ColⅠ and Col Ⅲ increased significantly (P<0.01). The content of ColⅠand ColⅢ could be inhibited by different density of Rutin observably (P<0.01).③RT-PCR result showed that, PI3K mRNA expression of M group increasedsignificantly (P <0.01), but PI3K mRNA expression of RHgroup were significantly lowerthan M group (P <0.01).④Acording to the results of Western Blotting, in the Ang II-induced cardiacfibroblast after treating24h, the expression level of TGF-β1, CTGF, PI3K, and Akt in Mgroup increased significantly (P<0.01). In R groups the expression of TGF-β1, CTGF, andAkt lowered observably (P <0.01); PI3K protein also has inhibited (P <0.01or P <0.05),but in RHthe effect was significantly (P <0.01).2. In vivo experience:①The result of HE, in the Iso induced cardiac fibroblast anddifferent densities of Rutin after treating24h, the degree of fibrosis was improved.②Immunohistochemical results show that the Col I and Col Ⅲ expression of Mgroup was higher than C group significantly (P <0.01), and that of Val and RM, RHwasobviously lower than M group (P <0.01) excepted the Col Ⅲ expression of RL.③According to the result of RT-PCR, the PI3K mRNA expression of M group raisedsignificantly (P <0.01), and that of RLand RM was significantly lower than M group (P <0.01).④Through the measure of TGF-β1, CTGF, PI3K and p-Akt/Akt in the Iso-inducedrats by Western Blotting, we found that these proteins expression were up-regulatedsignificantly (P<0.01). TGF-β1, CTGF and PI3K expression of RMand RH group weresignificantly lower than M group, and RHgroup had obviously inhibitory effect onactivation of Akt (P<0.01).Conclusion:1. Rutin could inhibit deposition of ColⅠ and Col Ⅲ by down-regulating the expression of TGF-β1and CTGF. So it could improve the lesion extent ofmyocardial fibrosis in rats induced by AngⅡ and Iso and delay the development ofmyocardial fibrosis. The effect had dose-dependence.2. Rutin could reserve the myocardial fibrosis via reducing the mRNA and proteinexpression of relative moleculars in PI3K/Akt pathway.