| Objectives This study was designed to explore the inhibition effect of matrine on isoproterenol-induced rats myocardial hypertrophy, and research the cytokines and the proteins expression of Rho A/ROCK signal pathway in rats’ myocardial tissues with ISO-induced myocardial hypertrophy in order to clarify the molecular mechanisms of matrine inhibits acute decompensatory cardiac hypertrophy.The cytokines include NO, ADMA, IGF-1, b FGF, TGF-β1, col1a1, col3a1 and e NOS.Methods Rats were injected subcutaneously ISO(85mg·kg-1) establish rat model of acute decompensatory cardiac hypertrophy, the rats were randomly divided into the normal control group, the matrine treatment groups(50, 100 and 200 mg·kg-1), the isoproterenol(ISO) model group, the matrine-alone group and the propranolol group(10 mg·kg-1). 24 h after the two-day drug administration, blood was collected from the right carotid artery, serum samples were separated and stored at-20°C for assay. The ratios of left ventricular weight to heart weight(LVW/HW) were determined, and heart morphometry was assessed. Hemodynamic(Heart rate, LVSP, LVd P/dtmax, and LVd P/dtmin) and biochemical parameters were measured. The cardiac apexes from each rat were excised and fixed in 10% neutral buffered formalin solution and the tissues were embedded in paraffin, sectioned and stained with hematoxylin and eosin(H&E) to observe Pathology structure change. The enzyme-linked immunosorbent assay was performed for NO, ADMA, IGF-1, b FGF, TGF-β1, and Cn T-I levels in the serum. Western blotting taken advantage of determine p-e NOS/e NOS and Rho A/ ROCK expressions in the left ventricular tissues. Real-time fluorescent quantitative RT-PCR was utilized to determine IGF-1, b FGF, TGF-β1, col1a1, col3a1 m RNAs expressions.Key findings 1. LVW/HW and HemodynamicCompared with the normal group, the ISO group rat’s LVW/HW,LV d P/dtmax and LV d P/dtmin were significantly decreased, In contrast,the matrine(50,100 and 200mg·kg-1)groups and the propranolol group(10mg·kg-1) LVW/HW,LV d P/dtmax and LV d P/dtmin were significantly increased in ISO-induced rats. But there was no significant difference between the matrine-alone group and the normal group in those aspects. 2. Histopathology of the myocardium(HE)The histopathological study showed that matrine(50,100 and 200mg·kg-1)and propranolol group(10 mg·kg-1) could attenuated abnormalities of cardiomyocyte hypertrophy,myocardial fibrosis,infiltration of inflammatory cells, interstitial edema and other abnormalities in ISO-induced acute decompensatory cardiac hypertrophy rat. 3. Effects of matrine on serum ADMA, IGF-1, TGF-β1, c Tn-I, b FGF, NO levels(ELISA)In comparison with the normal group, the serum NO, IGF-1 and TGF-β1 levels in the ISO group were decreased.While the matrine( 50,100 and 200mg·kg-1)and propranolol group(10 mg·kg-1) were remarkably increased on the serum NO, IGF-1 and TGF-β1 levels in ISO-induced acute decompensatory cardiac hypertrophy rat. Compared with the ISO group, the serum ADMA and Cn T-I levels in the ISO group were increased. While the matrine(50,100 and 200mg·kg-1)and propranolol group(10 mg·kg-1) were markedly reduced on the serum ADMA and Cn T-I levels in ISO-induced acute decompensatory cardiac hypertrophy rat. In contrast, those between the matrine-alone group and the normal group showed no significant difference. There were no significant difference on the serum b FGF level among all groups. 4. Effects of matrine on left ventricular e NOS, p-e NOS, Rho A and ROCK1 proteins(Western blot)Western blot results demonstrated: compared with the normal group, the protein levels of p-e NOS(Ser1177) and e NOS significantly reduced, and the protein levels of Rho A and ROCK1 increased significantly in myocardial tissue of the ISO-induced acute cardiac hypertrophy.In contrast, the matrine(100 and 200mg·kg-1) were remarkably increased the protein levels of p-e NOS(Ser1177) and e NOS, and decreased the protein levels of Rho A and ROCK1 in the ISO-induced acute cardiac hypertrophy rat. The matrine(50 mg·kg-1)did not significantly influenced the expression of p-e NOS(Ser1177), e NOS, ROCK1 and Rho A proteins in the ISO-induced acute cardiac hypertrophy rat. In contrast,there were no significant difference in the e NOS, p-e NOS, Rho A and ROCK1 protein levels between the matrine-alone group and the normal group. The propranolol group(10 mg·kg-1) remarkably increased the protein levels of p-e NOS(Ser1177) and e NOS, and decreased the protein levels of Rho A and ROCK1 in the ISO-induced acute cardiac hypertrophy rat. 5. Effects of matrine on left ventricular IGF-1, TGF-β1, b FGF, col1a1, col3a1 m RNA(RT-PCR)Real-time fluorescent quantitative RT-PCR results showed: compared with the normal group, the expression of IGF-1, TGF-β1, b FGF, col1a1, col3a1 m RNAs significantly reduced in myocardial tissue of the ISO-induced acute cardiac hypertrophy. In contrast, the matrine(50, 100 and 200mg·kg-1) were remarkably increased the expression of IGF-1, TGF-β1, b FGF m RNAs and the matrine(100 mg·kg-1) were remarkably increased the expression of col1a1, col3a1 m RNAs in the ISO-induced acute cardiac hypertrophy rat. In contrast, The propranolol group(10 mg·kg-1) and the matrine(200 mg·kg-1)-alone group,the expression of IGF-1, TGF-β1, b FGF, col1a1, col3a1 m RNAs showed no significant difference in myocardial tissue of the ISO-induced acute cardiac hypertrophy.Conclusions Oral administration of matrine to rats attenuated ISO-induced myocardial hypertrophy. Matrine molecular mechanism related to the regulation of the upstream of IGF-1, TGF-β1and b FGF expressions, and the downstream of NO, ADMA and e NOS expressions via Rho A/ROCK1 signaling pathway. |
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