CAJ | 학술논문

目的探讨可分化为肾小管上皮细胞的脂肪间充质干细胞(ADMSC)对大鼠缺血再灌注诱导的急性肾损伤(AKI)的治疗作用. 方法 2011年6月至2012年3月选取雄性SD大鼠8只,体质量60～80 g.取大鼠腹股沟区脂肪组织获取原代MDMSC.将慢病毒转染至培养第2代的ADMSC.再选取雄性SD大鼠40只,体质量180～ 220 g.使用无损伤性动脉夹夹闭双侧肾蒂40 min,构建缺血再灌注诱导的AKI模型.将大鼠分为移植组和对照组,每组20只.移植组将2×106个慢病毒转染的ADMSC重悬于100 μl PBS后,分别注射到左肾实质的上、中、下极;对照组注射100 μl PBS.于移植细胞后第3、28天,分别取两组各5只大鼠的左肾标本行HE染色,观察肾组织的损伤情况;TUNEL染色法和PCNA染色法检测肾细胞的凋亡和增殖情况.于建模后第1、3、14、28天检测大鼠的血肌酐(SCr)和尿素氮(BUN). 结果移植细胞后第3天,HE染色检查示移植组的肾小管损伤评分为2.0分,低于对照组的3.4分,差异有统计学意义(P＜0.05).TUNEL和PCNA染色法检测移植组细胞凋亡平均数为30个,对照组为55个;移植组细胞增殖平均数为35个,对照组为10个,差异均有统计学意义(P＜0.05).移植细胞后第1、3、14、28天移植组SCr分别为40、32、26、26 μmol/L,对照组分别为70、58、38、37 μmol/L;移植组BUN分别为15、13、10、7 mmol/L,对照组分别为24、20、13、10mmol/L,差异均有统计学意义(P＜0.05). 结论 ADMSC对大鼠的AKI具有明显的修复作用.
목적 탐토가분화위신소관상피세포적지방간충질간세포(ADMSC)대대서결혈재관주유도적급성신손상(AKI)적치료작용. 방법 2011년6월지2012년3월선취웅성SD대서8지,체질량60～80 g.취대서복고구구지방조직획취원대MDMSC.장만병독전염지배양제2대적ADMSC.재선취웅성SD대서40지,체질량180～ 220 g.사용무손상성동맥협협폐쌍측신체40 min,구건결혈재관주유도적AKI모형.장대서분위이식조화대조조,매조20지.이식조장2×106개만병독전염적ADMSC중현우100 μl PBS후,분별주사도좌신실질적상、중、하겁;대조조주사100 μl PBS.우이식세포후제3、28천,분별취량조각5지대서적좌신표본행HE염색,관찰신조직적손상정황;TUNEL염색법화PCNA염색법검측신세포적조망화증식정황.우건모후제1、3、14、28천검측대서적혈기항(SCr)화뇨소담(BUN). 결과 이식세포후제3천,HE염색검사시이식조적신소관손상평분위2.0분,저우대조조적3.4분,차이유통계학의의(P＜0.05).TUNEL화PCNA염색법검측이식조세포조망평균수위30개,대조조위55개;이식조세포증식평균수위35개,대조조위10개,차이균유통계학의의(P＜0.05).이식세포후제1、3、14、28천이식조SCr분별위40、32、26、26 μmol/L,대조조분별위70、58、38、37 μmol/L;이식조BUN분별위15、13、10、7 mmol/L,대조조분별위24、20、13、10mmol/L,차이균유통계학의의(P＜0.05). 결론 ADMSC대대서적AKI구유명현적수복작용.
Objective To study the mechanism of adipose-derived mesenchymal stem cell (ADMSC) in treating rats ischemia/reperfusion (I/R) induced acute kidney injury (AKI).Methods From June 2011 to March 2012,AKI was induced in 40 male SD rats (weight 180-220 g) by clamping bilateral renal pedicles for 40 minutes.Another 8 SD rats (weight 60-80 g) were used to obtain the primary ADMSC from inguinal fat tissue.After being transferred by lentivirus,those cells were cultured for transplantation until passage two.Animals with AKI were then randomly treated by intraparenchymally injecting ADMSC/PBS solutions (ADMSC/PBS group,n =20) or PBS solutions only (PBS group,n =20) (2× 106 cells,100 μl).During injection,the solutions were injected into the upper,middle and lower part of left kidney.The HE staining from 5 rats in each group was used to detect the histological injury at 3 days and 4 weeks after injection,respectively.The apoptosis and proliferation of host renal cells were evaluated using TUNEL staining and PCNA staining.The serum levels of SCr and BUN in animals were measured at day 1,3,14 and 28 after injection.Results HE staining showed ADMSC/PBS group got a lower injury score compared with that in PBS group at 3 days and 4 weeks,respectively (2.0 vs.3.4,1.3 vs.2.6,P＜0.05).In the result of TUNEL staining,the mean number of apoptosis cells was 30 in ADMSC/PBS group and 55 in PBS group.In terms of PCNA staining,the mean number of proliferative cells was 35 in ADMSC/PBS group and 10 in PBS group.All those results suggested that ADMSC could significantly reduce apoptosis and increase proliferation of renal cell (P＜0.05,repectively).The levels of serum SCr in ADMSC/PBS group were lower than those in PBS groups at day 1,3,14 and 28,after injection,respectively (40 vs.70 μmol/L,32 vs.58 μmol/L,26 vs.38 μ mol/L,26 vs.37 μmol/L; P＜0.05).The similar results were shown in the levels of serum BUN between the 2 groups (15 vs.24 mmol/L,13 vs.20 mmol/L,10 vs.13 mmol/L,7 vs.10 mmol/L; P＜0.05).Conclusion ADMSC could repair AKI after acute I/R injury.