摘要
目的:观察并分析大鼠心脏移植急性排斥反应与心肌细胞凋亡及心脏组织Fas及FasL表达的相关性。方法:实验于2006-01/07在南方医科大学南方医院动物实验中心完成。选用SPF级健康雄性SD,Wistar大鼠各36只,制作Wistar至SD大鼠Ono心脏移植模型,将36只SD受体大鼠按随机数字表法分为4组,即术后1,3,5,7d组,每组9只。①用QRT-PCR法检测大鼠术后1,3,5,7天心脏组织Fas,FasLmRNA的表达量,其中Fas蛋白表达于心脏组织中,FasL蛋白表达在活化的T细胞上。②应用脱氧核糖核苷酸末端转移酶介导的缺口末端标记技术检测心肌细胞的凋亡细胞,阳性细胞胞核呈现棕黄色。凋亡指数=凋亡细胞数/总细胞数×100%。③根据常规病理切片进行急性排斥反应分级,计0￣4级,0级:无排斥,心肌组织正常,心肌间质无炎细胞浸润;4级:弥漫性多类型细胞浸润,明显心肌间质水肿、出血和心肌坏死。结果:36只心脏移植模型大鼠全部进入结果分析,无脱失。①大鼠心脏移植术后随时间推移排斥反应逐渐加重,术后第7天排斥反应最重。②随急性排斥反应的加重,凋亡指数、FasL的表达均逐渐增高(0级:1.626±0.677,0.198±0.023;4级:11.404±1.274,0.408±0.051),发生4级排斥反应大鼠的心肌细胞凋亡指数、心脏组织FasLmRNA的表达量最高;而FasmRNA的表达量保持不变。③心脏移植术后受体大鼠发生急性排斥反应的程度与心肌细胞凋亡指数及心脏组织FasLmRNA的表达成正相关(r=0.863,0.880,P<0.01);急性排斥反应程度与FasmRNA表达不相关(r=0.032,P>0.05)。结论:心脏移植术后受体大鼠发生急性排斥反应的程度与心肌细胞凋亡指数及心脏组织FasLmRNA的表达成正相关,与FasmRNA表达不相关,提示可能是FasL阳性浸润细胞-活化的T细胞诱导Fas阳性心肌细胞凋亡从而介导移植排斥反应。
AIM： To observe and analyze the correlation of apoptosis, expressions of Fas and FasL with acute rejection in transplantation in rats. METHODS： The experiment was performed at the Animal Experimental Center, Nanfang Hospital, Southern Medical University from January to July 2006. Totally 36 SD and 36 Wistar healthy male rats of SPF grade were selected to make Ono heterotopic heart transplantation models from Wistar to SD rats. The 36 SD receptors were divided into 4 groups by random digital table method, namely 1-day operation group, 3-day operation group, 5-day operation group and 7-day operation group, 9 in each group. （1）QRT-PCR method was used to measure the expression of Fas and FasLmRNA in heart tissues at days 1, 3, 5 and 7 after operation. The expression of Fas expressed in heart tissues, while expression of FasL expressed in activated T-cells. （2）Apoptotic cells of grafted heart were detected by TUNEL. Positive cells were buffy. Apoptotic index was equal to number of apeptotic cells/total number of cells×100%. （3）Acute rejection grading was performed according to routinely pathological section, namely 0-4 grades, grade 0 as no rejection, with normal myocardium, and there was no inflammatory cells in myocardial interstitium; grade 4 as diffuse multiple cell infiltration, obvious interstitial edema, haemorrhage and myocardiolysis. RESULTS： Totally 36 rats of heart transplantation were involved in the result analysis, no drop-out. （1）After heart transplantation, the acute rejection became gradually severe with the prolongation of time, and reached the peak at day 7 after operation. （2）With the increasing of acute rejection, apoptotic index and FasL expression were increased gradually （grade 0：1.626±0.677,0.198±0.023 ;grade 4：11.404±1.274,0.408±0.051 ）. Apoptotic index and FasLmRNA expression were the peak in rats with grade 4 rejection. But the FasmRNA expression did not change. （3）After heart transplantation, degree of acute rejection was positively correlated wit
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第21期4074-4076,共3页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
广东省自然科学基金课题(990386)
广东省医学研究基金课题(A1999326)~~
