生物膜包裹间充质干细胞对甘油所致急性肾损伤小鼠肾的保护作用

Protective effect of mesenchymal stem cells wrapped in biological membrane on acute kidney injury of mice induced by glycerol

目的研究生物膜（一种异种脱细胞真皮基质）包裹间充质干细胞对甘油所致急性肾损伤（acute kidney injury,AKI）小鼠肾的保护作用。方法健康雄性C57BL/6小鼠（8-12周龄,体质量20-25g）40只,适应性喂养1周后,随机分为4组,每组各10只：正常对照组,双肾包膜剥离手术＋0.9%氯化钠注射液（Sham＋NS）;AKI模型组,双肾包膜剥离手术＋甘油注射（RM）;生物膜组,生物膜包裹无肾包膜双肾＋甘油注射（RM＋BM）;生物膜包裹间充质干细胞组,生物膜包裹间充质干细胞包裹无包膜双肾＋甘油注射（RM＋BM＋MSCs）。观察各组小鼠一般状态、血肌酐、尿素氮指标变化及肾组织学变化。结果 RM组、RM＋BM组与Sham＋NS组比较,小鼠出现精神萎靡、反应迟钝及肉眼血尿,进食进水量明显减少。RM＋BM＋MSCs组小鼠精神萎靡、反应迟钝及肉眼血尿的状况较RM组和RM＋BM组有所缓解。RM组小鼠与Sham＋NS组比较,血肌酐、尿素氮明显升高[（221.75±24.60）μmol/L vs（20.64±2.68）μmol/L,P〈0.01;（87.31±13.90）mmol/L vs（10.71±1.52）mmol/L,P〈0.01]。RM＋BM组小鼠的血肌酐、尿素氮较RM组无统计学差异[（206.68±21.75）μmol/L vs（221.75±24.60）μmol/L,P〉0.05;（87.62±11.23）mmol/L vs（87.31±13.90）mmol/L,P〉0.05]。RM＋BM＋MSCs组小鼠的血肌酐、尿素氮较RM组和RM＋BM组明显改善[（34.30±6.69）μmol/L vs（221.75±24.60）μmol/L、（206.68±21.75）μmol/L,P〈0.01;（12.32±1.57）mmol/L vs（87.31±13.90）mmol/L、（87.62±11.23）mmol/L,P〈0.01]。RM组和RM＋BM组肉眼可见肾大,PAS染色显示肾小管上皮细胞肿胀、变性、刷状缘消失和部分上皮细胞坏死,伴有肌红蛋白管型形成。RM＋BM＋MSCs组肾大不明显,肾损伤较RM组和RM＋BM组明显减轻,肾小管内管型减少,上皮细胞坏死减轻,可见不连续的刷状缘。结论生物膜包裹间充质干细胞对甘油所致急性肾损伤小鼠肾具有保护作用。

Objective To study the protective effect of mesenchymal stem cells（MSCs） wrapped in biological membrane（an acellular dermal matrix allograft） on acute kidney injury（AKI） of mice induced by glycerol. Methods Forty healthy male C57BL/6 mice（age of 8-12 weeks, weight of 20-25 g） were fed adaptively for 1 week, and they were randomly divided into four groups including： normal control group with renal capsular dissection surgery plus 0.9% Na Cl solution injection（Sham＋NS group, n=10）, AKI model group with renal capsular dissection surgery plus glycerol injection [RM（Rhabdomyolysis） group, n=10], biological membrane group with biological membrane wrapping up renal without capsule plus glycerol injection [RM＋BM（biological membrane） group, n=10] and MSCs wrapped in biological membrane group with mesenchymal stem cells（MSCs） wrapped in biological membrane packaging renal without capsule plus glycerol injection group（RM＋BM＋MSCs group, n=10）. The general status, indexes of renal function including serum creatinine and urea nitrogen as well as renal histopathological changes of each group were observed.Results Compared with Sham＋NS group, mice in RM and RM＋BM group showed symptoms of depression, macroscopic haematuria and poor appetite, while the mental state, appetite and haematuria of mice in RM＋BM＋MSCs group were better than RM group and RM＋BM group. Compared with Sham＋NS group, the serum creatinine and urea nitrogen of mice in RM group increased significantly [（221.75±24.60）μmol/L vs（20.64±2.68） μmol/L, P〈0.01;（87.31±13.90） mmol/L vs（10.71±1.52） mmol/L, P〈0.01], while it had no statistically significant different between RM group and RM＋BM group [（206.68±21.75） μmol/L vs（221.75±24.60） μmol/L, P〉0.05;（87.62±11.23） mmol/L vs（87.31±13.90） mmol/L, P〉0.05], and it showed significant improvement in RM＋BM＋MSCs group when compared with RM group and RM＋BM group [（34.30±6.69） μmol/L vs（221.75±24.60） μmol/L and（206.68±21.75） μmol/L, P〈0.01;（12.32±1.57） mmol/L vs（87.31±13.90） mmol/L and（87.62±11.23） mmol/L, P〈0.01]. Visible kidney enlargement was seen in RM group and RM ＋ BM group accompanied by swelling and degeneration in the renal tubular epithelial cells, disappearance of brush border, necrosis of part of the epithelial cells and formation of myoglobin tube in PAS staining. Compared with RM group and RM＋BM group, kidney enlargement was not obvious with slightly damage in RM＋BM＋MSCs group. The number of tube and necrosis reduced, and discontinuous brush border was seen in PAS staining. Conclusion MSCs wrapped in biological membrane shows protective effect on acute kidney injury of mice induced by glycerol.