摘要
目的探讨间充质干细胞外泌体上调Mir-21调控血红素氧合酶(HO-1)机制促进椎间盘退变的修复研究。方法新西兰健康大白兔36只随机数字法分成3组,Blank组(正常大白兔)、Model组(椎间盘退变大白兔)和Mir-21组(椎间盘退变大白兔+间充质干细胞外泌体),通过建模处理、外泌体提存、免疫组织化学染色、Rt-PCR检测、HE染色、流式细胞术检测,观察外泌体形态、对比分析HO-1蛋白阳性率在椎间盘退变中的表达,Mir-21、HO-1和BACH1在大白兔椎间盘组织中的表达,大白兔椎间盘组织形态学变化,白兔退变髓核细胞凋亡能力情况。结果Blank组HO-1蛋白阳性率为(62.23±4.1)%,细胞颗粒呈棕黄色,Model组的蛋白阳性率为(8.96±2.4)%,明显低于Blank组(P<0.05),Mir-21组蛋白阳性率为(42.38±3.46)%,细胞颗粒呈淡黄色,显著高于Model组低于Blank组(P<0.05);Model组的Mir-21、HO-1表达水平明显低于Blank组和Mir-21组(P<0.05),Mir-21组的Mir-21、HO-1表达水平显著低于Blank组(P<0.05)。Model组的BACH1水平明显高于Blank组和Mir-21组(P<0.05),Mir-21组的BACH1水平显著高于Blank组(P<0.05);Blank组兔髓核内基质比较高,髓核细胞颜色较深,周边胞浆颜色较淡,Model组通过建模后髓核细胞消失,与Blank组相比基质密度比较低,髓核细胞难见(P<0.05),Mir-21组通过Mir-21转染后,与Model组相比髓核内基质明显升高,密度染色加深,髓核细胞出现,并且染色加深(P<0.05);Model组和Mir-21组的细胞凋亡能力明显低于Blank组(P<0.05),Mir-21组细胞凋亡能力显著高于Model组(P<0.05)。结论间充质干细胞外泌体上调Mir-21,抑制BACH1来促进HO-1机制修复椎间盘退变的能力,对退变组织细胞具有促凋亡能力。
Objective To investigate the role of mesenchymal stem cell exosomes in up-regulating Mir-21 regulation of HO-1 mechanism to promote the repair of intervertebral disc degeneration.Methods New Zealand healthy white rabbits were randomly divided into 3 groups:Blank group(normal white rabbit),Model group(intervertebral disc degeneration white rabbit),and Mir-21 group(intervertebral disc degeneration white rabbit+mesenchymal stem cell exocytosis)Body model,by exosome extraction,immunochemical tissue staining,Rt-PCR detection,HE staining,flow cytometry detection,observation of exosome morphology,comparative analysis of HO-1 protein positive rate in intervertebral disc degeneration The expression of Mir-21,HO-1 and BACH1 in the intervertebral disc tissue of the white rabbit,the morphological changes of the intervertebral disc of the white rabbit,and the apoptosis ability of the degenerated nucleus pulposus cells in the white rabbit.Results The positive rate of HO-1 protein in Blank group was(62.23±4.1)%,and the cell granules were brownish yellow.The positive rate of protein in Model group was(8.96±2.4)%,which was significantly lower than that in Blank group(P<0.05).The positive rate was(42.38±3.46)%,and the cell granules were light yellow,which was significantly higher than that of the model group(P<0.05).The expression levels of Mir-21 and HO-1 in the Model group were significantly lower than those in the Blank group and Mir-21.The expression levels of Mir-21 and HO-1 in Mir-21 group were significantly lower than those in Blank group(P<0.05).The BACH1 level in the Model group was significantly higher than that in the Blank group and the Mir-21 group(P<0.05).The BACH1 level in the Mir-21 group was significantly higher than that in the Blank group(P<0.05).The rabbit nucleus in the Blank group had a higher matrix.The color of the nucleated cells was darker and the surrounding cytoplasm was lighter.The model group disappeared after modeling.Compared with the Blank group,the matrix density was lower,the nucleus pulposus
作者
龚东亮
付文芹
GONG Dong⁃liang;FU Wen⁃qin(Department of Orthopaedics,Qingpu Branch,Zhong⁃shan Hospital Affiliated to Fudan University,Shanghai 201799,China)
出处
《解剖学研究》
CAS
2020年第2期140-145,共6页
Anatomy Research
