骨髓间充质干细胞对大鼠痛风肾的修复作用  

Effect of bone marrow mesenchymal stromal cells on gout kidney

作　　者：李娜[1] 贾晓静 冯杏[1] 韩艳 赵丽君[4] 崔建军[4] Li Na;Jia Xiaojing;Feng Xing;Han Yan;Zhao Lijun;Cui Jianjun(1Department of Pediatric, Shanxi Medical University, Taiyuan 030001,China;Department of Pediatric, Shenzhen City People′s Hospital, Shenzhen 518020, China;Department of Neonatal Intensive Care Unit, Shanxi Provincial Children′s Hospital, Xian 710061, China;Department of Nephrology, Shanxi Provincial Children′s Hospital, Taiyuan 030013, China)

机构地区：[1]山西医科大学儿科医学系,太原030001 [2]深圳市人民医院儿科,518020 [3]陕西省儿童医院新生儿监护室,西安710061 [4]山西省儿童医院肾脏内科,太原030013

出　　处：《中华临床医师杂志（电子版）》2017年第11期1894-1901,共8页Chinese Journal of Clinicians(Electronic Edition)

基　　金：山西省科技攻关项目(20100311102-1)

摘　　要：目的探讨经鼠尾静脉移植的骨髓间充质干细胞(BM-MSCs)对大鼠痛风肾的影响,具体为延缓上皮到间叶组织转化(EMT)、促进肾脏细胞分化和生长,以及抗氧化应激等方面的作用。并进一步研究其相应的机制。方法幼年雄性Wistar大鼠随机分为正常组和造模组,造模组采用腺嘌呤200 mg/(kg·d)灌胃4周制作痛风肾大鼠模型,造模成功24 h后,造模组再次被随机分为3组,分别为磷酸盐缓冲液模型组和BM-MSCs治疗组,BM-MSCs治疗组经鼠尾静脉移植入骨髓间充质干细胞(BM-MSCs是经密度梯度离心法结合贴壁筛选法在体外分离培养而成),同时磷酸盐缓冲液(PBS)模型组经鼠尾静脉注入相同量的PBS。注射6周后,收集大鼠的血液和尿液标本用来测定血肌酐、尿素氮和24 h尿蛋白含量;对大鼠肾石蜡标本切片进行H-E染色、糖原染色和马松染色,观察并半定量评分评价各组肾病理情况;免疫组织化学法测定大鼠肾石蜡标本切片中的转化生长因子β1(TGF-β1)的表达水平,Western-Blot法测定肾组织中P38蛋白、P-P38蛋白、硫氧还蛋白还原酶1(Trx R1)的表达情况。数据经过正态性及方差齐性检验后,正常组和造模组两两比较采用LSD-t检验,3组之间比较采用单因素差分析。P<0.05表示差异有统计学意义。结果经腺嘌呤诱导制作的痛风肾大鼠相比正常组大鼠出现明显的肾功能降低,大量蛋白尿以及肾间质和肾小管尿酸结晶沉积,肾间质纤维化、炎症细胞浸润、肾小管上皮细胞坏死、肾小球硬化等,提示痛风肾模型制造成功,并出现了慢性肾功能衰竭;BM-MSCs治疗组较PBS模型组而言,血肌酐水平降低[(88.90±7.89)μmol/Lvs.(117.40±6.13)μmol/L],尿素氮降低[(7.85±0.88)mmol/L vs.(10.97±1.03)mmol/L],24 h尿蛋白减少[(27.72±4.90)mg vs.(54.66±6.72)mg],TGF-β1表达减少[(11.00±2.28)个vs.(20.67±1.63)个],P-P38/P38比值降低[(0.31±0.09)μmol/L vs.(0.50±0.13)μmol/L],Trx R1/β-actin�Objective The objective of this study was to investigate the effects of the intravenous transplantation of bone marrow mesenchymal stromal cells(BM-MSCs)on alleviating epithelial to mesenchymal transition(EMT)and promoting renal cell differentiation and growth and anti-oxidative stress in rats with gout kidney.Furthermore,the corresponding mechanisms were explored.Material and Methods A rat model with gout kidney was established by adenine inducing for4weeks.Immature male Wistar rats were randomly divided into control group,model group and treatment group.The BM-MSCs treatment group rats were injected with BM-MSCs via tail vein24h after the successful modeling,whereas the phosphate-buffered saline model group rats were injected with phosphate-buffered saline(PBS).Six weeks later,urine and blood were collected to assess24-hour proteinuria,serum creatinine(Scr)and blood urea nitrogen(BUN).Immunohistochemistry was perfomed to determine the expression of transforming growth factor-β1(TGF-β1).We used Western blot to determine protein expression of P-P38/P38and selenium-containing enzyme thioredoxin reductase1(TrxR1)in renal tissues.Statistical analysis of differences between the control group and the model group was performed using t-test,and the date of three groups(the control group,the BM-MSCs treatment group,the PBS model group)was calculated using analysis of variance(ANOVA).A value of P<0.05was considered statistically significant.Results Rats with gout kidney induced by adenine were more likely to have mass proteinuria,deterioration of renal function and the histopathologic injury in the kidney,which implied that the gout kidney might evolve into chronic renal failure(CRF).Compared with the PBS model group,the level of indicators in BM-MSCs treatment group was signficantly lower including Scr[(88.90±7.89μmol/L)vs.(117.40±6.13μmol/L)],BUN[(7.85±0.88mmol/L)vs.(10.97±1.03mmol/L)],24hours proteinuria[(27.72±4.90mg)vs.(54.66±6.72mg)],TGF-β1expression[(11.00±2.28)vs.(20.67±1.63)]and the ratio of P-P38and

关 键 词：痛风肾 慢性肾功能衰竭 骨髓间充质干细胞 上皮至间叶转化 转化生长因子Β1 肿瘤坏死因子-α/P-P38 硫氧还蛋白还原酶1 

分 类 号：R589.7[医药卫生—内分泌] R692[医药卫生—内科学]