摘要
目的:利用反义脱氧寡核苷酸技术探讨组蛋白脱乙酰基酶2(HDAC2)对心肌细胞肥大的促进作用,为探寻心肌细胞肥大的发病机理及其防治提供一新的思路。方法:常规方法培养原代心肌细胞,利用血管紧张素Ⅱ(AngⅡ)刺激心肌细胞制成心肌细胞肥大的体外模型,应用Fugene 6转染HDAC2-反义脱氧寡核苷酸进行干预。以逆转录聚合酶链反应(RT-PCR)观察HDAC2和β肌球蛋白重链(β-MHC)mRNA的表达;通过相差显微镜观察心肌细胞的形态和面积;利用免疫组织化学法检测心肌细胞HDAC2和c-fos蛋白的表达。结果:与对照组相比,肥大组心肌细胞面积,HDAC2 mRNA和β-MHC mRNA表达均增加(P均<0.01);HDAC2蛋白和c-fos蛋白表达增加(P<0.01)。利用Fugene 6将HDAC2-反义脱氧寡核苷酸转染入心肌细胞,HDAC2-反义脱氧寡核苷酸组心肌细胞面积较肥大组减小,HDAC2 mRNA和β-MHC mRNA表达减少,HDAC2蛋白表达亦减少(P均<0.05)。Fugene 6组,错义组与肥大组变化相似。结论:AngⅡ致心肌细胞肥大过程中伴有HDAC2的mRNA和蛋白表达增加,应用HDAC2反义脱氧寡核苷酸后可使之下降,说明HDAC2参与了心肌细胞的肥大机制。
Objective： To observe the inhibitory effect of histone deacetylase 2--antisense oligonucleotide on cardiocyte hypertrophy induced by angiotensin Ⅱ. Methods： The cultured cardiocytes divided into control group, Fugene group （treated with Ang Ⅱ and Fugene）, cardiocyte hypertrophy group （treated with Ang Ⅱ to induce hypertrophy）, HDAC2--ASODN group [given with histone deacetylase 2 （HDAC2） --antisense oligonucleotide （ASODN） and Ang Ⅱ] and HDAC2--missence oligonucleotide （MSODN） group （given with HDAC2, MSODN and Ang Ⅱ）. The morphologic changes of cardiocytes were observed under the contrast phase microscope. The mRNA levels of HDAC2 and β-MHC were examined by real-time PCR method. The protein expressions of HDAC2 and c-fos, was examined by immunohistochemistry method. Results： Compared with control group, the cardiocytes enlarged （P〈0.01） under the contrast phase microscope; the mRNA levels of HDAC2 and β-MHC and the protein expressions of HDAC2 and C-los increased （P〈0.01） in hypertrophic cardiocytes; but given by HDAC2--ASODN, these indexes decreased （P〈0.05）. There was no significant difference among Fugene 6 group and cardiocyte hypertrophy group and HDAC2-- MSODN group. Conclusion： The results means that HDAC2 may play a role in genesis of cardiocyte hypertrophy. HDAC2 ASODN may be a new drug to treat cardiocyte hypertrophy.
出处
《心血管康复医学杂志》
CAS
2009年第1期13-16,22,共5页
Chinese Journal of Cardiovascular Rehabilitation Medicine
基金
黑龙江省教育厅课题(11511208)
