移植同种异体骨髓间充质干细胞对兔椎间盘退变早期的干预实验研究  

Experimental study on the restraining effects of intervertebral discs degeneration of early stage by transplantation of allogeneic bone marrow mesenchymal stem cell

作　　者：白亦光[1] 陈巧玲[2] 刘康[3] 肖东琴[3] 罗栩伟[1] 林涛[1] 杨泽龙[1] 冯刚[3] BAI Yi-guang;CHEN Qiao-ling;LIU Kang;XIAO Dong-qin;LUO Xu-wei;LIN Tao;YANG Ze-long;FENG Gang(Departments of Orthopaedics,Nanchong Central Hospital,the Second Clinical Institute of North Sichuan Medical College,Nanchong 637000,Sichuan,China;Departments of Oncology,Nanchong Central Hospital,the Second Clinical Institute of North Sichuan Medical College,Nanchong 637000,Sichuan,China;Institute of Tissue Engineering and Stem Cells,Nanchong Central Hospital,the Second Clinical Institute of North Sichuan Medical College,Nanchong 637000,Sichuan,China)

机构地区：[1]川北医学院第二临床医学院·南充市中心医院骨科,四川南充637000 [2]川北医学院第二临床医学院·南充市中心医院肿瘤科,四川南充637000 [3]川北医学院第二临床医学院·南充市中心医院组织工程与干细胞研究所,四川南充637000

出　　处：《川北医学院学报》2020年第1期1-5,13,共6页Journal of North Sichuan Medical College

基　　金：国家自然科学基金(81171472、81071270、30872614);四川省南充市校科技战略合作专项(18SXHZ0539);四川省卫计委课题(16PJ202)。

摘　　要：目的:在诱导椎间盘退变的早期移植同种异体兔骨髓间充质干细胞(BMSC),观察BMSC对退变椎间盘的修复效果。方法:体外分离、培养BMSC,并通过组织化学染色、免疫组织化学和免疫荧光染色对其进行鉴定。采用穿刺抽吸法建立兔椎间盘退行性变模型,造模同期进行细胞移植治疗。27只5月龄日本大耳白兔随机分为正常对照组(NC组)、BMSC移植组(BMSC组)、DMEM培养基组(DMEM组),每组各9只。每只实验动物的L 3-4、L 4-5和L 5-6作为实验干预椎间盘。术后4、8、12周分别利用标准化T2加权像信号强度(%ST2WI)、病理组织学、退变分数、免疫组化染色、免疫荧光染色以及二型胶原(CollagenⅡ)和蛋白聚糖(aggrecan)mRNA表达水平评价修复效果。结果:术后12周,DMEM组的%ST2WI值、CollagenⅡ和aggrecan的mRNA表达量均低于其它各组(P<0.05),NC组和BMSC组比较,差异无统计学意义(P>0.05);NC组和BMSC组退变分数低于DMEM组(P<0.05),NC组和BMSC组比较,差异无统计学意义(P>0.05);NC组、BMSC组的CollagenⅡ免疫组化和aggrecan免疫荧光染色均显示强阳性,而DMEM组随着时间的推移阳性逐渐减弱。结论:在退变早期移植同种异体的兔BMSC能有效抑制椎间盘的退变。Objective:The aim of this study was to observe on the repair effect of rabbits degenerative intervertebral disc on early stage with bone marrow mesenchymal stem cell(BMSC)transplantation.Methods:BMSC was isolated and cultured in vitro,and identified with histochemical staining,immunochemical staining and Immunofluorescence staining.The intervertebral discs degeneration animal model was constructed by the conventional punctured method and at the same time BMSC was transplanted into the rabbit s model of disc degeneration.27 Japanese white rabbits of 5-month-old were randomly divided into three groups:normal control group(NC group),BMSC transplantation group(BMSC group)and DMEM medium group(DMEM group),9 cases in each group.L 3-4,L 4-5 and L 5-6 intervertebral discs were set as samples of experiment in every rabbit.In 4,8 and 12 weeks after the transplantation,repair effect of rabbits degenerative intervertebral discs were evaluated by standard T2-weighted image ratio(%ST2WI),histology,degeneration score,immunochemical staining,Immunofluorescence staining,and mRNA expression levels of collagen II and aggrecan.Results:In 12 weeks after cell-transplantation,standard T2-weighted image ratio(%ST2WI),mRNA expression levels of collagen II and aggrecan for DMEM group were significantly lower than other groups(P<0.05).There was no difference between NC group and BMSC group(P>0.05).The degeneration score for the NC and BMSC groups was significantly lower than that of the DMEM group(P<0.05).There was no significant difference between NC and BMSC groups(P>0.05).Immunofluorescence staining of CollagenⅡand aggrecan indicated obviously positive in NC and BMSC groups,and that of DMEM group gradually weakened with the change of time.Conclusion:In the early stage of degeneration,BMSC can effectively inhibit the degeneration of intervertebral disc.

关 键 词：骨髓间充质干细胞 椎间盘退行性变 干细胞移植 

分 类 号：R394.2[医药卫生—医学遗传学]