CAJ | 학술논문

目的探讨DNA甲基转移酶3A(DNMT3A)和羟脯氨酸( Hyp)在盐酸异丙肾上腺素( ISO)诱导的大鼠心肌纤维化中的表达及相关性研究。方法将40只雄性SD大鼠随机等分为正常对照组和模型组。模型组给予ISO [5 mg/kg ·d)],正常对照组皮下注射给予等量生理盐水,连续7 d后处死大鼠获取血液标本,并取心肌组织。同时,测定心重指数( HW/BW)、左心室质量指数( LVW/BW)；采用ELISA法检测血清中Ⅰ型胶原和Ⅲ型胶原的含量；HE染色和Masson染色法观察心肌纤维化程度；Western blot法测定 DNMT3A和α平滑肌肌动蛋白(α-SMA)的表达；紫外分光光度法测定心肌组织中Hyp的含量。结果与正常对照组比较,模型组HW/BW、LVW/BW明显增加；模型组血清标本中Ⅰ型胶原和Ⅲ型胶原的含量较正常对照组明显增加；HE染色和Masson染色显示模型组心肌组织出现明显的胶原纤维增生；Western blot法检测显示模型组DNMT3A和α-SMA的表达明显高于正常对照组(P<0.05,P<0.01)；同时,紫外分光光度法检测显示模型组心肌组织中Hyp含量明显增高( P<0.05)；DNMT3A和Hyp在大鼠心肌纤维化组织中的表达呈正相关(r=0.675,P<0.05)。结论DNMT3A表达上调在心肌纤维化的形成过程中起重要促进作用,可能与 Hyp升高有一定的相关性。
목적탐토DNA갑기전이매3A(DNMT3A)화간포안산( Hyp)재염산이병신상선소( ISO)유도적대서심기섬유화중적표체급상관성연구。방법장40지웅성SD대서수궤등분위정상대조조화모형조。모형조급여ISO [5 mg/kg ·d)],정상대조조피하주사급여등량생리염수,련속7 d후처사대서획취혈액표본,병취심기조직。동시,측정심중지수( HW/BW)、좌심실질량지수( LVW/BW)；채용ELISA법검측혈청중Ⅰ형효원화Ⅲ형효원적함량；HE염색화Masson염색법관찰심기섬유화정도；Western blot법측정 DNMT3A화α평활기기동단백(α-SMA)적표체；자외분광광도법측정심기조직중Hyp적함량。결과여정상대조조비교,모형조HW/BW、LVW/BW명현증가；모형조혈청표본중Ⅰ형효원화Ⅲ형효원적함량교정상대조조명현증가；HE염색화Masson염색현시모형조심기조직출현명현적효원섬유증생；Western blot법검측현시모형조DNMT3A화α-SMA적표체명현고우정상대조조(P<0.05,P<0.01)；동시,자외분광광도법검측현시모형조심기조직중Hyp함량명현증고( P<0.05)；DNMT3A화Hyp재대서심기섬유화조직중적표체정정상관(r=0.675,P<0.05)。결론DNMT3A표체상조재심기섬유화적형성과정중기중요촉진작용,가능여 Hyp승고유일정적상관성。
Objective To investigate the expression and correlation of DNA methyltransferase 3A( DNMT3A) and Hyp of isoprenaline( ISO) induced myocardial fibrosis in rats. Methods 40 male SD rats were randomly divided into normal control group and model group. The model group was treated subcutaneously with 5 mg/( kg·d) iso-prenaline daily for 7 days, but the normal control group was given the same amount of normal saline at the same time. Rats were executed to obtain blood samples and myocardial tissue. And HW/BW, LVW/BW were tested. ELISA was used to detect the content of type Ⅰ collagen and type Ⅲ collagen in serum. The degree of myocardial fibrosis was observed with HE staining and Masson staining method. The expression of DNMT3A and α-SMA was tested by Western blot. The content of hydroxproline in myocardial tissue was measured by UV spectrophotometry. Results HW/BW and LVW/BW increased significantly in the model group compared with the normal control group. The content of type Ⅰ collagen and type Ⅲ collagen in serum increased significantly in the model group compared with the normal control group. HE and Masson staining showed myocardial tissue in model group ap-peared obviously collagen fiber hyperplasia. The expression of DNMT3A andα-SMA in the model group was higher than the normal control group. UV spectrophotometric detection showed the content of Hyp in myocardial tissue in the model group increased obviously. The expression of DNMT3A andα-SMA in myocardial fibrosis tissue was pos-itively correlated(r=0.675, P<0.05). Conclusion DNMT3A upregulation plays an important role in the myo-cardial fibrosis, and Hyp may increase a certain correlation.