慢性肾衰竭模型中透明质酸和CD_(44)对骨髓间充质干细胞归巢及修复作用的影响  

Effect of Hyaluronic Acid and CD_(44) on Homing and Improving Renal Function of Mesenchymal Stem Cells in A Rat Chronic Renal Failure Model

作　　者：边晓慧[1,2] 何平[2] 孙广萍[2] 孙立[1] 李德天[2] 冯江敏[1] 

机构地区：[1]中国医科大学附属第一医院肾内科,辽宁省沈阳市110001 [2]中国医科大学附属盛京医院肾内科

出　　处：《中国全科医学》2014年第18期2103-2108,共6页Chinese General Practice

基　　金：辽宁省自然科学基金(20072095)

摘　　要：目的探讨慢性肾衰竭模型中透明质酸(HA)与CD44的相互作用对骨髓间充质干细胞(MSC)归巢和修复受损肾脏能力的影响。方法根据随机数字表法将32只雄性SD大鼠分为:假手术组(n=8)、模型组(n=8)、MSC组(n=8)和MSC/抗CD44组(n=8)。采用间隔1周的两步法建立大鼠5/6肾切除模型,假手术组只进行麻醉和肾被膜剥离,而不进行肾切除术。第二次术后第8周分别给予尾静脉注射1×107/ml MSC、1 ml磷酸盐缓冲液(PBS)、1×107/ml MSC、1×107/ml抗CD44抗体OX50预孵的MSC。治疗4周后处死大鼠,留取血标本和残肾组织。检测血肌酐和尿素氮水平;过碘酸雪夫(PAS)染色和Masson染色后观察病理改变,应用肾小球硬化指数和肾小管间质损伤指数来判定肾小球和肾小管间质损伤程度;免疫组化法观察残肾组织中巨噬细胞和肌成纤维细胞的变化;免疫荧光法追踪归巢的MSC;免疫组化法和免疫印迹法观察HA的表达;双荧光法观察MSC与HA的共定位关系。结果与假手术组相比,模型组大鼠的血肌酐、尿素氮、肾小球硬化指数、肾小管间质损伤指数升高(P<0.05),肾脏病理改变主要为肾小球肥大、局灶节段性肾小球硬化,炎性细胞浸润和肾间质纤维化,包括巨噬细胞和肌成纤维细胞数量明显增加;与模型组相比,MSC组、MSC/抗CD44组肌酐、尿素氮、肾小球硬化指数、肾小管间质损伤指数均降低(P<0.05)。假手术组肾皮质几乎不表达HA,而模型组的肾皮质大量表达HA。免疫荧光法显示,治疗4周后,假手术组未发现有归巢的MSC,MSC组的残肾组织中有较多的MSC,MSC/抗CD44组归巢的MSC减少。双免疫荧光结果显示较多MSC归巢至HA表达丰富的区域。结论 HA和CD44的相互作用影响MSC的归巢能力,但不影响MSC修复受损肾脏的能力。Objective To explore the effect of hyaluronic acid（ HA）and CD44 on the homing and improving renal function of Mesenchymal Stem Cells（ MSC）in a rat chronic renal failure model. Methods 32 grade clean adult male SD rats were randomly divided into 4 groups：sham operation group,renal failure group,MSC group,MSC/anti -CD44 group,8 in each group. Establishment of rat remnant kidney model：after one week adaptive breeding,all cut right kidney first,then cut 2/3 left kidney 1 week later. But in sham operation group,after anesthesia,rats were only given renal capsule stripping. 8 weeks after model establishment,1 ml MSC（1 × 10^7/ml）,1 ml PBS,1 ml MSC（1× 10^7/ml）and 1 ml MSC（1 ×10^7/ml）incuba-ted with OX50（anti CD44 antibody）were injected via tail vein in every group respectively. After treatment of 4 weeks,all rats were killed and blood sample and kidney tissue were collected. Serum Cr and BUN were tested. Pathological changes of kidney were assessed by PAS and Masson stain. Glomerulosclerosis index and renal tubular mesenchymal damage index were calculated by semiquantitative method. Tracing MSC using immunofluoresent assay. Double fluorescence immunostain of MSC and HA was used in kidney sections of rat. Expression of HA was assayed by immunohistochemical staining and western blot. Results Compared with sham operation group,renal failure group displayed significantly elevated Cr,BUN,glomerulosclerosis index and renal tubular mesenchymal damage index,and the difference was significant（P〈0. 05）. The results of PAS and Masson staining indicated that compared to sham operation group major pathological changes in residule kidney tissues were glomerular hypertrophy, cloudy swelling of renal epithelial cells,focal mesangial cell proliferation,widened mesangial matrix,focal glassy degeneration of glomerular capillary,renal interstitial inflammatory cell infiltration,focal interstitial fibrosis. MSC and MSC/anti-CD44 treatments could alleviate the above pathological changes in MSC group and MS

关 键 词：肾功能衰竭 慢性 骨髓间充质干细胞 透明质酸 CD44 归巢 

分 类 号：R692.5[医药卫生—泌尿科学]