全反式维甲酸调控Kruppel因子5在预防移植静脉术后狭窄的作用及其机制  

All trans retinoic acid mediated kruppel like factor 5 during the prevention of post transplant vein stenosis

作　　者：余咏潮 朱培培 王国坤 徐志云 唐昊 Yu Yongchao;Zhu Peipei;Wang Guokun;Xu Zhiyun;Tang Hao(Department of Cardiothoracic Surgery,Shanghai Changhai Hospital,Shanghai 200082,China;Shanghai Institute of Electrical Engineering,Shanghai 201306,China)

机构地区：[1]上海长海医院胸心外科,200082 [2]上海电机学院,201306

出　　处：《中华实验外科杂志》2022年第5期834-838,共5页Chinese Journal of Experimental Surgery

基　　金：2021年海军军医大学基础医学项目(2021MS04)。

摘　　要：目的探讨全反式维甲酸(ATRA)调控Kruppel因子5(Klf5)在预防移植静脉术后狭窄的作用及机制。方法建立新西兰大白兔(海军军医大学动物实验中心提供)静脉动脉化动物模型,按完全随机分为造模组(A、B、C组分别为饲养2、4、8周)和ATRA组(D、E、F组分别为饲养2、4、8周,鼻饲ATRA10 mg/d)、空白对照组(G组),各6只。分别获取移植静脉标本,行苏木精-伊红染色及免疫组化检验。建立人脐静脉平滑肌细胞(SMC)KLF5过表达细胞模型,以ATRA干预,划痕实验、细胞增殖实验(CCK-8)明确SMC增殖及迁移情况;免疫共沉淀明确ATRA对Klf5-RARa结合的阻断作用。两组间计量资料采用两独立样本t检验,多组间计量资料采用单因素方差分析,组间两两比较采用LSD-t检验方法。结果各组管径:A组(107.58±18.11)μm,B组(144.65±26.10)μm,C组(160.28±28.06)μm,D组(78.42±9.00)μm,E组(102.75±16.47)μm,F组(117.47±38.06)μm,G组(35.73±6.04)μm。组间比较,建模组各组(A、B、C各组)明显高于空白对照组(G组)(t=5.81、8.81、10.08,P<0.01),同期建模组各组明显高于ATRA组(t=2.06、2.96、3.03,P<0.05)。以染色指数法计算免疫组化结果,建模各组细胞核增殖抗原(Ki-67,2周3.07±0.64,4周3.67±0.81,8周1.93±0.41)表达明显高于对照组(t=6.93、9.37、4.16,P<0.01),也高于同期ATRA组(2周2.87±0.52,4周3.60±1.21,8周2.10±0.24,t=4.91、6.66、2.14,P<0.05)。实验组Klf5表达(2周4.43±0.70,4周5.67±1.18,8周3.03±0.98)明显高于对照组(t=7.27、9.09、3.83,P<0.01),建模组与ATRA组间KLF5(2周4.43±0.70,4周5.67±1.18,8周3.03±0.98)表达差异无统计学意义(t=0.49、0.17、0.42,P>0.05)。CCK-8实验:72 h Klf5组吸光度(A)值(1.54±0.20)高于其余3组(t=5.62、5.84、8.31,P<0.01),Klf5+ATRA组(1.02±0.14)高于ARTA组(0.80±0.07,t=2.47,P<0.05),对照组(1.04±0.13)高于ATRA组(t=2.70,P<0.05)。划痕实验:以迁移前后划痕距离的比值,klf5组0.098±0.006,对照组0.404±0.009,ATRA组0.597±0.014,Klf5Objective To study the role and mechanism of all trans retinoic acid(ATRA)regulating Kruppel factor 5(KLF5)in the prevention of stenosis after vein transplantation.Methods New Zealand white rabbits(provided by the Animal Experimental Center of Naval Military Medical University)were randomly divided into two groups:model group(groups A,B,C for 2,4 and 8 weeks,respectively),ATRA group(groups D,E,F for 2,4 and 8 weeks respectively,nasal ATRA 10 mg/day)and blank control group(group G).The transplanted vein samples were obtained and tested by hematoxylin eosin staining and immunohistochemistry.The KLF5 overexpression cell model of human umbilical vein smooth muscle cells(SMCs)was established.The proliferation and migration of SMCs were determined by ATRA intervention,scratch test and cell proliferation test(CCK-8).Immunocoprecipitation confirmed the blocking effect of ATRA on KLF5 rara binding.The measurement data between the two groups were analyzed by two independent sample t-test,the measurement data between multiple groups were processed by one-way ANOVA,and the comparison between the two groups was done by LSD-t test.Results The diameter in groups A,B,C,D,E,F and G was(107.58±18.11),(144.65±26.1),(160.28±28.06),(78.42±9.00),(102.75±16.47),(117.47±38.06)and(35.73±6.04)μm respectively.The diameter in the modeling groups was significantly greater than in the blank control group(t=5.81,8.81,10.08,P<0.01),and that in the modeling groups was significantly greater than in the ATRA groups(t=2.06,2.96,3.03,P<0.05).The expression of Ki67(3.07±0.64 at 2nd week,3.67±0.81 at 4th week and 1.93±0.41 at 8th week)in the modeling groups was significantly higher than that in the blank control group(reference index 1)(t=6.93,9.37 and 4.16,P<0.01),and also higher than that in the ATRA group(2.87±0.52 at 2nd week,3.60±1.21 at 4th week and 2.1±0.24 at 8th week,t=4.91,6.66 and 2.14,P<0.05).The expression of KLF5 in the experimental group(4.43±0.70 at 2nd week,5.67±1.18 at 4th week and 3.03±0.98 at 8th week)was significa

关 键 词：冠状动脉旁路移植术 全反式维甲酸 Kruppel因子5 预防 移植静脉再狭窄 

分 类 号：R654.3[医药卫生—外科学]