组织因子途径抑制物2对SD乳鼠心房成纤维细胞和心房肌细胞功能的影响  

Impact of tissue factor pathway inhibitor-2 on atrial fibroblasts and atrial cardiomyocytes

作　　者：洪晋 黄玉起 白中乐[1] 朱记法[1] 张振宇[2] 赵文[3] 刘献志[2] 董建增[1,4] HONG Jin;HUANG Yuqi;BAI Zhongle;ZHU Jifa;ZHANG Zhenyu;ZHAO Wen;LIU Xianzhi;DONG Jianzeng(Department of Cardiology,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Department of Neurosurgery,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Pharmacy School,Zhengzhou University,Zhengzhou 450001;Department of Cardiology,Beijing Anzhen Hospital,Capital Medical University,Beijing 100029)

机构地区：[1]郑州大学第一附属医院心血管内科,郑州450052 [2]郑州大学第一附属医院神经外科,郑州450052 [3]郑州大学药学院,郑州450001 [4]首都医科大学附属北京安贞医院心内科,北京100029

出　　处：《郑州大学学报（医学版）》2022年第2期176-181,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金项目(81702465,U1804172);中国博士后科学基金项目(2019M652591);河南省自然科学基金项目(212300410257)。

摘　　要：目的:探讨组织因子途径抑制物2(TFPI-2)对心房成纤维细胞和心房肌细胞功能的影响及相关分子机制。方法:采用ELISA法检测15例心房颤动(AF)患者和15名正常对照血清TFPI-2水平。分离、培养并鉴定SD乳鼠心房成纤维细胞和心房肌细胞。通过CCK-8实验检测0、50、100、200μg/L重组TFPI-2蛋白(rTFPI-2)处理24、48 h对心房成纤维细胞增殖能力的影响。采用Transwell小室实验检测100μg/L rTFPI-2处理24 h对心房成纤维细胞迁移能力的影响,采用Annexin V-FITC/PI双染法检测rTFPI-2处理48 h对心房成纤维细胞和心房肌细胞凋亡的影响,采用Western blot法检测细胞中相关蛋白的表达。结果:AF患者血清TFPI-2水平低于正常对照(P<0.05)。成功分离、培养SD乳鼠心房成纤维细胞和心房肌细胞并鉴定。rTFPI-2可抑制心房成纤维细胞增殖、迁移,并促进其凋亡(P<0.05),但rTFPI-2不增加心房肌细胞凋亡(P>0.05)。rTFPI-2下调心房成纤维细胞p-Erk1/2及MMP-9和MMP-2表达,上调PARP表达(P<0.05),但对心房肌细胞PARP表达无显著影响(P>0.05);rTFPI-2抑制两种细胞Ⅲ型、Ⅰ型胶原表达(P<0.05)。结论:TFPI-2可抑制心房成纤维细胞增殖和迁移,促进心房成纤维细胞凋亡,但不增加心房肌细胞凋亡,TFPI-2能抑制两种细胞胶原合成;TFPI-2具有抑制心房纤维化的潜在价值。Aim:To explore the impact of tissue factor pathway inhibitor-2(TFPI-2)on atrial fibroblasts and atrial cardiomyocytes and the molecular mechanism.Methods:Serum TFPI-2 level in atrial fibrillation(AF)patients(n=15)and controls(n=15)was determined by ELISA.Atrial fibroblasts and atrial cardiomyocytes from neonatal SD rats were isolated,cultured and identified.The influence of 0,50,100,200μg/L recombinant TFPI-2 protein(rTFPI-2)on cell proliferation of atrial fibroblasts was detected via CCK-8 assay after 24,48 hours culture.The effects of 100μg/L rTFPI-2 on atrial fibroblasts migration after 24 hours culture were inspected by Transwell chamber,the effects on atrial fibroblasts′and atrial cardiomyocytes′apoptosis after 48 hours culture were detected by Annexin V-FITC/PI double staining,and associated proteins in cells were tested by Western blot.Results:Serum TFPI-2 level was lower in AF patients than that in controls.The atrial fibroblasts and atrial cardiomyocytes from neonatal SD rats were successfully isolated and identified.rTFPI-2 could suppress cell proliferation,migration,promote apoptosis of atrial fibroblasts(P<0.05),but did not promote apoptosis of atrial cardiomyocytes(P>0.05).Although rTFPI-2 down-regulated p-Erk1/2,MMP-9 and MMP-2 expression and up-regulated PARP expression in atrial fibroblasts(P<0.05),it did not change PARP expression in atrial cardiomyocytes(P>0.05).CollagenⅢand collagenⅠin both cells were suppressed by rTFPI-2(P<0.05).Conclusion:TFPI-2 could suppress proliferation and migration of atrial fibroblasts,and promote apoptosis,but would not increase atrial cardiomyocytes apoptosis;TFPI-2 inhibits collagen expression in both cells.TFPI-2 may be effective in the process of anti-atrial fibrosis.

关 键 词：组织因子途径抑制物2 心房成纤维细胞 心房肌细胞 心房纤维化 

分 类 号：R541.75[医药卫生—心血管疾病] R542.23[医药卫生—内科学]