CAJ | 학술논문

目的研究骨髓源干细胞( BMDSC)移植对进展性肾小球硬化模型大鼠的作用,同时观察移植的BMDSC能否分化为肾脏内皮细胞和促进肾小球毛细血管修复再生.方法把携带增强型绿色荧光蛋白( EGFP)转基因SD大鼠的BMDSC移植于EGFP-SD大鼠制成嵌合性大鼠.注射抗Thy-1.1抗体,30 min后再予切除右侧肾脏制成进展性肾小球硬化模型.按是否给予BMDSC治疗分为非治疗组及治疗组,每组10只,实验期限12周.在不同时间点,检测肾功能,并进行光镜、免疫荧光病理检查.观察移植后大鼠情况,以及移植的BMDSC是否分化为肾脏内皮细胞和促进肾小球毛细血管修复再生.结果12周时,非治疗组大鼠仅有3只存活,余7只分别死于抗体注射后第2、7、9、11周；治疗组大鼠9只存活,仅1只死于第10周.与非治疗组比较,治疗组肾功能明显改善[BUN( 43.55±29.06)比(76.58±66.19) mmol/L,Scr (138.79±75.14)比(233.38± 164.43) μmol/L].第3、7、14、28天时两组24h尿蛋白量差异无统计学意义,而在第42、56、84天时,治疗组显著低于非治疗组.非治疗组可见严重系膜细胞和基质增生及肾小球硬化,而治疗组上述增生与硬化明显减轻；非治疗组肾小球硬化指数和系膜细胞增生指数显著高于治疗组,差异有统计学意义.治疗组肾小球可见多个BMDSC,细胞数显著多于非治疗组和正常嵌合鼠.用鼠内皮细胞标记抗体RECA-1进行双重免疫荧光染色显示,治疗组有更多BMDSC参与了肾小球内皮细胞的修复及再生,并且显著促进了肾小球毛细血管的修复再生.结论 BMDSC静脉输入对进展性肾小球硬化有治疗作用,BMDSC可分化成内皮细胞,并可促进肾小球毛细血管的修复再生.
목적 연구골수원간세포( BMDSC)이식대진전성신소구경화모형대서적작용,동시관찰이식적BMDSC능부분화위신장내피세포화촉진신소구모세혈관수복재생.방법파휴대증강형록색형광단백( EGFP)전기인SD대서적BMDSC이식우EGFP-SD대서제성감합성대서.주사항Thy-1.1항체,30 min후재여절제우측신장제성진전성신소구경화모형.안시부급여BMDSC치료분위비치료조급치료조,매조10지,실험기한12주.재불동시간점,검측신공능,병진행광경、면역형광병리검사.관찰이식후대서정황,이급이식적BMDSC시부분화위신장내피세포화촉진신소구모세혈관수복재생.결과12주시,비치료조대서부유3지존활,여7지분별사우항체주사후제2、7、9、11주；치료조대서9지존활,부1지사우제10주.여비치료조비교,치료조신공능명현개선[BUN( 43.55±29.06)비(76.58±66.19) mmol/L,Scr (138.79±75.14)비(233.38± 164.43) μmol/L].제3、7、14、28천시량조24h뇨단백량차이무통계학의의,이재제42、56、84천시,치료조현저저우비치료조.비치료조가견엄중계막세포화기질증생급신소구경화,이치료조상술증생여경화명현감경；비치료조신소구경화지수화계막세포증생지수현저고우치료조,차이유통계학의의.치료조신소구가견다개BMDSC,세포수현저다우비치료조화정상감합서.용서내피세포표기항체RECA-1진행쌍중면역형광염색현시,치료조유경다BMDSC삼여료신소구내피세포적수복급재생,병차현저촉진료신소구모세혈관적수복재생.결론 BMDSC정맥수입대진전성신소구경화유치료작용,BMDSC가분화성내피세포,병가촉진신소구모세혈관적수복재생.
Objective To investigate the protective role of bone marrow-derived stem cells (BMDSCs) in progressive glomerulosclerosis rats,and to observe whether BMDSCs promote glomenlar repair and regeneration.Methods Progressive glomerulosclerosis was induced in enhanced green fluorescent protein (EGFP) bone marrow chimeric rats by injecting with anti-Thy1.1 antibody,followed by unilateral nephrectomy.Subsequently,these rats were treated with either BMDSCs infusion (treatment group,10 rats) or phosphate-buffered saline (untreated group,1O rats).Renal function and histological alterations were examined at week 12 after Thy1.1 antibody injection.Repair and regeneration of glomerular endothelial cells was detected by immunofluorescence.Results Only 3 rats survived in untreated group,other 7 rats died at week 2,7,9,11 after antibody injection.In treatment group,9 rats survived at week 12,only 1rat died at week 10.The BUN and Scr were significantly lower in treatment group as compared to untreated group [BUN(43.55±29.06) vs (76.58±66.19) mmol/L,Scr (138.79±75.14) vs (233.38±164.43) μmol/L].Proteinuria was not significantly different between two groups at day 3,7,14 and 28,while it was significantly decreased at day 42,56 and 84 in treatment group compared with untreated group.Light microscopy showed that there was severe diffuse mesangial cell proliferation,mesangial matrix expansion and glomerulosclerosis in untreated group,and such changes were ameliorated in treatment group.The mesangial expansion index and glomerular sclerosis index in untreated group was significantly higher than those in treatment group.More BMDCs were recruited into the glomeruli and differentiated into glomerular endothelial cells in treatment group as compared with untreated group.Double immunofluorescence stain also demonstrated that BMDSCs infusion promoted glomerular capillary repair and regeneration.Conclusion BMDSCs infusion can improve renal function and histological changes,and promote the repair and regeneration of glomerular capillary.