中文摘要：

目的探讨脂肪间充质干细胞（ADSC）对烧伤脓毒症小鼠肾脏损伤的作用及相关机制。方法（1）取5只C57BL／6J小鼠双侧腹股沟脂肪组织，采用酶消化法、密度梯度离心法及贴壁法分离培养纯化，小鼠ADSC，取第3代细胞行形态学观察、测定细胞生长曲线，流式细胞仪检测细胞表面分子标记物，并行成骨及成脂肪细胞诱导分化鉴定，（2）另取37只C57BI．／6J小鼠，按随机数字表法分为正常埘照组、生理盐水组、ADSC组。正常对照组5只，其余2组各16只。生理盐水组及ADSC组小鼠造成背部15％TBSAm度烫伤后，创面中心注射铜绿假单胞菌菌液制成烧伤脓毒症小鼠模型，之后分别经尾静脉注射生理盐水和ADSC，均于伤后12、24、48、72h观察肾脏组织病理学变化、检测血肌酣和尿素氮水平，并采用实时荧光定量PCR法检测。肾脏组织TNF—a、IL-12、IL-10以及环氧合酶2（COX2）mRNA表达。正常对照组小鼠不行任何处理，但均行上述检测。对数据行多因素方蓐分析及LSD-t检验，结果（1）分离培养的细胞传至第3代时，形态均一与Fb相似，排列规则；CD90‘、CDl05’、CD34-、CD45细胞百分比均在90％以L；细胞可向成脂和成骨细胞分化；经签定，细胞为ADSC。（2）伤后12～72h，牛理盐水组小鼠肾小管和肾小球中中性粒细胞浸润逐渐增多，管腔受损、肾小球结卡勾被破坏。ADSC组小鼠伤后各时相点肾组织病理学损伤程度均轻于生理盐水组。生理盐水组各时卡fj点m肌酐、尿素氮水平均明显高于正常对照组（P值均小于0．01）；伤后12～72h．与生理盐水组比较，AI）SC组血肌酐、尿素氮水平均明显下降（P值均小于0．01）。与正常对照组比较，伤后24h生理盐水组及ADSC组TNF-a、IL-12mRNA表达量均升高（P值均小于0．05）。伤后2411，ADSC组TNF—dtuRNA水平（1．58±0．19）明最低于生理盐水组的3．36±0．

英文摘要：

Objective To investigate the effect of adipnse-derived stem cells （ADSC） on renal injury in mice with burn injury and sepsis and its underlying mechanism. Methods （ 1 ） Adipose tissue was collected from both inguinal regions of C57BL/6J mice to isolate, culture and purify A DSC through enzyme digestion, density gradient eentrifugation, and adherence method. Cells of the third passage were used in the experiment. The nmrphologie change in cells was obserw~d and the growth eurw of cells was determined. The expression of cell surface antigen phenotype was analyzed by flow cytometry, and the eells were identified by adipugenie and osteogenie differentiation. （2） Another 37 C57BL/6J mice were divided into normal eontrol group （ n =5）, saline group （ n =16）, and group ADSC （ n =16） according to the random number table. The mice in saline group and group ADSC were injected with Pseudomonas aeruginosa after being subjected to 15% TBSA full-thiekness burn on the back to reproduee septic burn model. Then the mice wereinjected with saline and ADSC through tail vein respectively. At post burn hour （PBH） 12, 24, 48, and 72, the pathological change in kidney tissue was observed, the levels of blood urea nitrogen and serum creatinine were determined, and the levels of TNF-a, IL-12, IL-10, and cyclooxygenase-2 （COX2） mRNA were determined with real-time fluorescence quantitative PCR in both groups. Above-mentioned indexes were also examined in the normal control group （without burn）. Data were processed with multifactor analysis of vari- ance and LSD- t test. Results （ 1 ） Cells in the third passage were orderly arranged with the shape similar to fibroblasts. The percentages of CD90 , CD105 , CD34- , and CD45 - cells were all above 90% . The cells could differentiate into osteoblasts and adipoeytes. The cells were identified to be ADSC. （2） From PBH 12 to PBH 72, the neutrophil infiltration gradually increased, and the structure of kidney tubules and glomeruli were deranged in saline