粒细胞集落刺激因子改善急性心肌梗死模型大鼠的心肌纤维化  

Granulocyte colony-stimulating factors improve myocardial fibrosis in rats with myocardial infarction

作　　者：殷婷婷 杜大勇[2] 蒋知新[2] 柳杨[2] 刘奇林 李运田 Yin Tingting;Du Dayong;Jiang Zhixin;Liu Yang;Liu Qilin;Li Yuntian(The Second School of Clinical Medicine,Southern Medical University,Guangzhou 510000,Guangdong Province,China;The 305th Hospital of Chinese PLA,Beijing 100000,China)

机构地区：[1]南方医科大学第二临床医学院,广东省广州市510000 [2]中国人民解放军第305医院心脏中心,北京市100000

出　　处：《中国组织工程研究》2022年第5期730-735,共6页Chinese Journal of Tissue Engineering Research

基　　金：全军冠心病诊疗中心建设资助项目(09GXB),项目负责人:李运田;国家自然科学基金资助项目(30971235),项目参与人:蒋知新。

摘　　要：背景:粒细胞集落刺激因子作为一种强效干细胞动员剂,在心肌梗死早期使用可以动员骨髓间充质干细胞至心肌梗死缺血区,调控细胞外基质代谢,抑制心肌纤维化。目的:观察粒细胞集落刺激因子对急性心肌梗死大鼠心肌纤维化的影响。方法:冠状动脉结扎法建立大鼠急性心肌梗死模型,建模成功的大鼠随机分为模型对照组及治疗组(n=7),另外取7只大鼠仅穿线,不结扎,作为空白对照组。治疗组于建模后24 h皮下注射粒细胞集落刺激因子[50μg/(kg·d)],模型对照组和空白对照组均于相同部位注射相应体积生理盐水,1次/d,共5 d。4周后,苏木精-伊红染色观察心肌组织形态学变化,Masson染色计算胶原容积分数,ELISA法检测血清Ⅰ型前胶原羧基端肽和Ⅲ型前胶原氨基端肽含量,免疫组织化学法和蛋白免疫印迹法(Western blot)检测大鼠心肌梗死边缘区基质金属蛋白酶诱导剂CD147及基质金属蛋白酶2、基质金属蛋白酶抑制剂2蛋白表达水平。结果与结论:(1)4周后,与空白对照组相比,模型对照组胶原容积分数明显增加(P <0.001),CD147、基质金属蛋白酶2蛋白水平明显升高(P <0.001),基质金属蛋白酶抑制剂2蛋白水平明显降低;(2)与模型对照组相比,治疗组胶原容积分数明显降低(P <0.001),CD147、基质金属蛋白酶2蛋白水平明显降低(P <0.001),基质金属蛋白酶抑制剂2蛋白水平明显增加(P <0.001);(3)与空白对照组相比,模型对照组的Ⅰ型前胶原羧基端肽、Ⅲ型前胶原氨基端肽表达水平明显增加(P <0.001);与模型对照组相比,治疗组的Ⅰ型前胶原羧基端肽、Ⅲ型前胶原氨基端肽表达水平明显降低(P <0.001);(4)提示粒细胞集落刺激因子可改善急性心肌梗死大鼠心肌纤维化,可能是通过下调CD147蛋白的表达水平,直接或间接调控基质金属蛋白酶2/基质金属蛋白酶抑制剂2比例平衡、改善细胞外基质代谢、抑制胶原沉积从BACKGROUND: As a powerful stem cell mobilization agent, granulocyte colony-stimulating factor can mobilize bone marrow mesenchymal stem cells to the ischemic area in the early stage of myocardial infarction, regulate extracellular matrix metabolism and inhibit myocardial fibrosis in the early stage of myocardial infarction.OBJECTIVE: To observe the effect of granulocyte colony-stimulating factor on myocardial fibrosis in rats with acute myocardial infarction.METHODS: Animal models of acute myocardial infarction were established in rats by coronary artery ligation. After successful modeling, the rats were randomly divided into two groups: model group(n=7) and treatment group(n=7). Another seven rats were randomly selected as blank control group without ligation. At 24 hours after modeling, the treatment group was subcutaneously injected with granulocyte colony-stimulating factor(50 μg/kg/d). Both the model group and the blank control group were injected with the corresponding volume of normal saline at the same site, once a day for 5 days. After 4 weeks, the morphological changes of myocardial tissue were observed by hematoxylin-eosin staining, and the collagen volume fraction was calculated by Masson staining. The levels of serum type I procollagen carboxyl terminal peptide and type III procollagen N-terminal peptide were detected by ELISA assay. The protein expressions of matrix metalloproteinase inducer(CD147), matrix metalloproteinase 2 and matrix metalloproteinase inhibitor 2 in the marginal zone of myocardial infarction were detected by immunohistochemistry and western blot. RESULTS AND CONCLUSION: After 4 weeks, the volume fraction of collagen was significantly higher in the model group than in the blank control group(P < 0.001), the protein levels of CD147 and matrix metalloproteinase 2 were significantly increased(P < 0.001), and the level of matrix metalloproteinase inhibitor 2 protein was significantly decreased. Compared with the model group, the collagen volume fraction in the treatment group decreas

关 键 词：心肌梗死 心肌纤维化 粒细胞集落刺激因子 CD147 基质金属蛋白酶2 基质金属蛋白酶抑制剂2 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]