移植脂肪干细胞抑制大鼠颈动脉损伤后新生内膜的异常增殖  

Transplanting adipose tissue-derived mesenchymal stem cells inhibits neointimal hyperplasia after carotid balloon injury in rats

作　　者：翁剑锐 许昌声 王华军 林志鸿[1,2] WENG Jian-rui;XU Chang-sheng;WANG Hua-jun;LIN Zhi-hong(Department of Emergency,the First Affiliated Hospital of Fujian Medical University;Fujian Hypertension Research Institute,Fuzhou,Fujian 350005,China)

机构地区：[1]福建医科大学附属第一医院急诊科 [2]福建省高血压研究所,福建福州350005

出　　处：《中华高血压杂志》2021年第12期1215-1221,共7页Chinese Journal of Hypertension

基　　金：福建省自然科学基金项目(2015J01456)。

摘　　要：目的探讨移植脂肪干细胞(ADMSC)对球囊损伤后大鼠颈动脉新生内膜异常增殖的影响及机制。方法45只雄性SD大鼠,随机分为3组:对照组(假手术,n=15),球囊损伤组(n=15),ADMSC移植组(n=15)。对照组仅予颈部切开缝合,球囊损伤组进行左侧颈总动脉球囊损伤,ADMSC组在血管损伤同时予以ADMSC移植治疗。在术后的第1、2、4周时处死大鼠,随机取第2、3、5、7天观察左侧颈总动脉,观察绿荧光蛋白标记的ADMSC在颈总动脉的定植情况;苏木精-伊红染色观察颈总动脉内膜增生;酶联免疫吸附试验法测定血清中一氧化氮的水平,逆转录聚合酶链反应检测颈总动脉内皮型一氧化氮合酶(eNOS)mRNA的表达;免疫印迹法测定颈总动脉细胞周期素依赖性激酶抑制蛋白p27^(kip1)蛋白、p21^(Cip1)蛋白以及增殖细胞核抗原(PCNA)蛋白的水平。结果球囊损伤4周后,SD大鼠颈总动脉内膜增生明显,血管壁腔比值(W/L)高于对照组(1.24±0.08比0.38±0.09,t=13.84,P<0.01);ADMSC移植后可定植于血管内膜,且显著抑制球囊损伤后新生内膜的异常增殖,血管W/L低于损伤组(0.63±0.05比1.24±0.08,t=-9.75,P<0.01)。球囊损伤后,与对照组比较,颈总动脉p27^(kip1)、p21^(Cip1)蛋白的表达水平呈时间依赖性增加,于术后4周达高峰;而球囊损伤后PCNA的表达水平在1周后达高峰,2周后降至正常。移植ADMSC增加了球囊损伤4周时颈总动脉p21^(Cip1)蛋白及p27^(kip1)蛋白的表达;同时降低了损伤后1周PCNA的表达。球囊损伤后4周,损伤血管eNOS mRNA表达及血清一氧化氮水平[(56.37±9.91)比(112.27±9.05)μmol/L,t=-7.66,P<0.05]降低;ADMSC移植后,损伤血管eNOS mRNA表达及血清一氧化氮水平[(75.83±3.67)比(56.37±9.91)μmol/L,t=2.96,P<0.05]增加。结论移植的ADMSC可以定植于损伤血管,抑制球囊损伤后颈动脉新生内膜的异常增殖,机制可能与上调p21^(Cip1),p27^(kip1),减少PCNA的表达,同时增加损伤血管eNOS mRNA的表达,Objective To investigate the effects and mechanism of transplanting adipose tissue-derived mesenchymal stem cells(ADMSC)on neointimal hyperplasia after carotid balloon injury in rats.Methods Forty-five male Sprague Dawley rats were randomly divided into three groups:sham group(control group,n=15),injured group(balloon injured,n=15),and ADMSC group(balloon injury and treatment with ADMSC,n=15).The balloon catheter was inserted into the left carotid to produce intimal injury in injured group and ADMSC group,while neck midline incision alone was performed in control group.After 1,2and 4weeks,rats were euthanized,and the injured carotid artery and normal carotid artery were used for HE-stained pathological study.Serum concentration of nitric oxide(NO)was detected by enzyme-linked immune sorbent assay(ELISA),mRNA expression of endothelial nitric oxide synthase(eNOS)was determined by reverse transcription-polymerase chain reaction(RT-PCR),and the protein expressions of p21^(Cip1),p27^(kip1) and proliferating cell nuclear antigen(PCNA)were determined respectively by Western blot.Results Enhanced green fluorescent protein positive cells were detected in the traumatized arterial segment within 1week after transplantation.Compared with control group,the wall-to-lumen area ratios(W/L)in injured group were increased significantly(1.24±0.08 vs 0.38±0.09,t=13.84,P<0.01).The W/L ratios in ADMSC group were lower than in injured group(0.63±0.05 vs 1.24±0.08,t=-9.75,P<0.01).The protein expressions of p21^(Cip1) and p27^(kip1) increased significantly in a time-dependent manner and reached peak level at 4thweek after balloon injury.Meanwhile,the protein expression of PCNA significantly increased and reached peak level at 1st-week after balloon injury.In comparison with injured group,ADMSC treatment significantly increased the protein expressions of p27^(kip1),p21^(Cip1)(P<0.05),and the expression of PCNA was significantly decreased after 1-week treatment with ADMSC(P<0.05).Compared with control group,the serum level of NO[(56.37

关 键 词：血管球囊损伤 脂肪干细胞 移植 新生内膜增殖 

分 类 号：R543.4[医药卫生—心血管疾病]