羊膜干细胞对严重烧伤大鼠早期心肌损伤的保护作用  

The protective effects of amniotic mesenchymal stem cells for rats early myocardial injury induced by severe burns

作　　者：牟欢[1] 刘洋[2] 刘剑 周新立[1] 强廷会[1] 杜兴国[1] 丁晨[1] 薛超[1] 

机构地区：[1]陕西省汉中市中心医院骨关节外科一病区,陕西汉中723000 [2]第四军医大学第一附属医院全军烧伤中心,陕西西安710032 [3]第四军医大学第一附属医院神经外科,陕西西安710032

出　　处：《中国医药导报》2017年第19期4-7,23,共5页China Medical Herald

基　　金：国家自然科学基金资助项目(81501666)

摘　　要：目的探讨羊膜干细胞(AMSCs)对严重烧伤大鼠早期心肌损害的作用及相关作用机制。方法采用酶消化法分离培养AMSCs,并用流式细胞仪对所分离培养细胞表面分子标志物进行检测,并进行体外诱导成脂、成骨分化鉴定。24只健康雄性SD大鼠,按照随机数字表法进行实验分组,分为单纯烧伤(burn)组、AMSCs组、假伤(sham)组。实验大鼠背部于98℃热水浴15 s,制成30%总体表面积Ⅲ度烧伤模型,伤后即刻burn组、AMSCs组大鼠分别腹腔注射0.9%氯化钠10 m L(50 m L/kg)行抗休克治疗,3 h后两组大鼠分别经尾静脉注射PBS和AMSCs;sham组仅对大鼠背部于37℃水浴15 s,模拟烧伤模型制作过程。48 h后采集三组大鼠腹主动脉血及心肌组织标本,采用ELISA法检测乳酸脱氢酶(LDH)、血清肌酸激酶(CK)的含量,RT-PCR法检测组织中凋亡相关因子(caspase-3),炎性相关因子TNF-α、IL-1β及抑炎因子IL-10 m RNA表达水平。对所得实验数据进行单因素方差分析、LSD-t检验。结果采用酶消化法培养的AMSCs传至第3代后检测均表达CD29、CD44、CD90、CD105,而CD34阳性率较低,可在体外向成骨成脂诱导分化。与sham组比较,burn组大鼠血清LDH、CK的含量显著增高(P<0.05),AMSCs组CK、LDH含量与burn组相比显著降低(P<0.05)。burn组大鼠心肌中TNF-α、caspase-3、IL-1βm RNA水平均高于sham组(P<0.05),AMSCs组心肌组织中TNF-α、caspase-3、IL-1βm RNA水平显著低于burn组(P<0.05)。burn组大鼠心肌组织中IL-10 m RNA表达量显著低于sham组(P<0.05),AMSCs组大鼠心肌组织中IL-10 m RNA表达量与burn组比较显著增高(P<0.05)。结论严重烧伤早期采用AMSCs治疗可显著降低心肌CK、LDH水平,减轻严重烧伤导致的心肌组织损伤,降低心肌细胞凋亡,减少炎症细胞因子IL-1β和TNF-α表达,促进抑炎细胞因子IL-10的表达水平,对严重烧伤大鼠心脏细胞损伤具有重要的保护作用。Objective To explore the effects and related mechanism of arnniotic mesenchymal stem cells （AMSCs） for rats myocardial injury induced by severe bums. Methods The AMSCs were obtained by the method of collagenasepancreatic enzyme digestion. Biological markers were assayed with flow cytometry. The activity of AMSCs was identi- fied using adipogenic and osteoplastic differentiation. 24 healthy male SD rats were randomly divided into sham group, burn group and AMSCs group. The dorsal skin of rats in burn group and AMSCs group were exposed to 98℃ water for 15 s which led to the third degree, 30% TBSA burns, while the rats in the sham group were exposed to 37℃ water for 15 s. Rats in burn group and AMSCs group were given 0.9% 10 mL （50 mL/kg） saline intraperitoneally immediately after burns. PBS or AMSCs were injected through caudal vein 3 hours later. 48 hours later, rats were sacrificed and the myocardium and blood from aorta abdominalis were collected. The levels of creatine kinase （CK）, lactic dehydrogenase （LDH） were detected by ELISA. The mRNA levels of caspase-3, TNF-α, IL-1β and IL-10 were detected by RT-PCR. The data were analyzed through A VONA andLSD-t test by SPSS. Results The positive rates of CD29, CD44, CD90, CD105 were very high in the third generation of AMSCs, however, the positive rate of CD34 was low. AMSCs underwent osteogenic and adipogenic differentiation after induction. The levels of CK and LDH in rats of the burn group were significantly increased compared with the sham group （P 〈 0.05）. The levels of CK and LDH in AMSCs group were significantly lower than those in the burn group （P 〈 0.05）. The mRNA levels of caspase-3, IL-115 and TNF-α in the myocardium of the burn group were significantly in- creased compared with the sham group （P 〈 0.05）. The mRNA levels of caspase-3, IL-1β and TNF-α in myocardium of the AMSCs group were significantly lower than those in the burn group （P 〈 0.05）. The mRNA level of IL-10 in the burn group was significantly l

关 键 词：烧伤 心肌损伤 羊膜干细胞 炎症因子 细胞凋亡 

分 类 号：R644[医药卫生—外科学]