Effect Of Hydrogen Sulfide On Myocardial Fibrosis In The Mice With Alcoholic Cardiomyopathy And The Primary Study Of Its Underlying Mechanism

Objective: This study takes the mice with alcoholic cardiomyopathy(ACM) that induced by chronic alcohol consumption as the research object. Then sodium hydrosulfide(Na HS), as hydrogen sulfide(H2S) honor, and DL-propargylglycine(PAG), an inhibitor of cystathionine-?-synthase(CSE) were respectively used to treat these mice, aiming to observe the effect of H2 S on myocardial fibrosis in the mice with alcoholic cardiomyopathy and explore its possible mechanism.Methods: Forty-four male Kunming mice were randomly divided into 4 groups: control group, model group, Na HS group, PAG group. Except the control group, other groups were established alcoholic cardiomyopathy model by feed with 4% alcohol for 12 weeks. At the beginning of starting the modeling, the mice in Na HS group were treated with Na HS(50 ?mol/kg per day) and the mice in PAG group were treated with PAG(40mg/kg per day) via intraperitonea(i.p.) injection. An equal volume of normal saline was injected in the control and model groups.12 weeks later, the body weight(BW)of all mice were measured before they were killed, then the hearts were taken out and the hearts weight(HW) were measured, and the ratios of heart weight to body weight(HW/BW) were calculated. Masson and VG staining were used to evaluate the cardiac collagen deposition. Immunohistochemistry was used to determining the expression of collagen ?. Transmission electron microscopy was used to observed autophagosomes. Western Blotting was used to determining the expression of autophagy-associated proteins Beclin 1, Atg3 and Atg7. Real-time quantitative PCR analysis was used to detect the level of mi R-21 and mi R-211.Results: 1.There was no statistical difference in BW, HW and HW /BW among each group(P>0.05). 2. The results of Masson and VG staining showed that, the collagen deposition in model group was more evident than control group. However, compared with model group, the collagen deposition was obviously alleviated in Na HS group. In contrast, the collagen deposition was further aggravated in PAG group than model group. 3. Immunohistochemistry result indicated that the expression of collagen?in model group was significantly higher than control group. However, compared with model group, the expression of collagen?was significantly reduced in Na HS group. In contrast, the expression of collagen?was further significantly increased in PAG group than model group. 4. Typical autophagosomes were observed in model and PAG group with transmission electron microscopy, while not observed in control and Na HS group. 5. The results of western blotting revealed that the expressions of Beclin 1, Atg3 and Atg7 in model group were significantly increased than control group(p?0.05). However, compared with model group, the expressions of Beclin 1, Atg3 and Atg7 were significantly down-regulated in Na HS group(p?0.05). In contrast, the expressions of Beclin 1 and Atg3 were further markedly increased in PAG group than model group(p?0.05). The expression of Atg7 was further increased in PAG group compared with model group although the difference was not significant(P>0.05). 6. The results of real-time quantitative PCR analysis showed that the expression level of mi R-21 and mi R-211 in model group were obviously elevated than control group(p?0.05). However, compared with model group, the expression level of mi R-21 and mi R-211 were obviously reduced(p?0.05)in Na HS group. In contrast, the level of mi R-211 was further significantly elevated in PAG group than model group(p?0.05). The expression of mi R-21 was further increased in PAG group compared with model group although the difference was not significant(P>0.05).Conclusion: H2 S can attenuate myocardial fibrosis in the mice with alcoholic cardiomyopathy, and its mechanism may involve inhibition of excessive activation of autophagy and down-regulating the expression of mi R-21 and mi R-211.