硫化氢对内皮素-1致心房肌细胞肥大的保护作用

The protective effect of hydrogen sulfide on atrial myocyte hypertrophy induced by endothelin-1

目的探讨硫化氢对内皮素-1(endothelin-1,ET-1)致心房肌细胞的保护作用,为未来房颤的药物治疗指导新思路。方法选取出生1~3 d的乳鼠,分离心房及心室肌细胞,通过化学酶消化法提取心房肌细胞,心房肌细胞培养3 d后经免疫细胞化学染色法鉴定,而后随机分为4组加入相应的药物:A组:对照组;B组:ET-1组(10-4~10-7mol/L);C组:ET-1+Na HS(ET-1:10-8mol/L,Na HS:10-4~10-7mol/L);D组:ET-1+ETRA(ET-1:10-8mol/L,ET-1受体拮抗剂:10-5~10-7mol/L)。PCR方法检测各组中CSE、Connexin40和Ca MKⅡmRNA的表达情况。结果经免疫细胞化学染色鉴定90%以上的心房肌细胞经α-肌动蛋白染色阳性。经PCR方法检测,与对照组比较,在B组中Connexin40和Ca MKⅡmRNA表达量显著增加(P<0.05),CSE mRNA表达量下降(P<0.05),尤其是ET-1浓度为10-8mol/L时(P<0.05);而在C组和D组中,Connexin40和Ca MKⅡmRNA表达量显著下降(P<0.05),而CSE mRNA表达量增加(P<0.05),尤其是浓度为10-6mol/L时(P<0.05)。结论ET-1可导致心房肌细胞肥大。H2S对ET-1所致的心房肌细胞肥大起到保护作用。

Objective To investigate the protective effect of H2 S in hypertrophy of atrial myocytes induced by endothelin-1(ET-1).Methods Atrial myocytes were isolated and cultured from 40 Sprague Dawley rats(1~3 days old).Atrial myocytes were identified with immunocytochemistry and drugs added to cell culture dishes after three days of cultivation and then randomly divided into four groups:①group A:control group;②group B:ET-1(10^-7~10^-9 mol/L);③group C:ET-1+NaHS(ET-1:10^-8 mol/L,NaHS:10^-4~10^-7 mol/L);④group D:ET-1+ETRA(ET-1:10^-8 mol/L,the ETRA,10^-5~10^-7 mol/L).Moreover,the expression of cystathionine-γ-lyase(CSE),Connexin 40 and CaMKⅡmRNA were examined with real-time polymerase chain reaction(PCR).Resultsα-actin antibody staining authenticated 90%of myocytes of atrium with immunocytochemistry.The expression of Connexin40 and CaMKⅡmRNA significantly increased in group B compared with the control group(P<0.05),especially in 10-8 mol/L;and significantly decreased in group C and D(P<0.05),particularly in 10-6 mol/L.Moreover,the expression of CSE mRNA significantly decreased in group B compared with the control group(P<0.05),especially in 10-8 mol/L;and significantly increased in group C and D(P<0.05),particularly in 10^-6 mol/L.Conclusion ET-1 could induce atrial myocyte in hypertrophic.H2 S could improve the hypertrophy of atrial myocytes induced by ET-1.