自体黄韧带干预TGF-β1/Smad3信号通路抑制兔硬膜外纤维瘢痕形成的实验研究  

Experimental study on the effect of autologous ligamenta flava on TGF-β1/Smad3 signaling pathway on epidural fiber scar formation in rabbits

作　　者：王鹏 张德宝 张杨 张海斌 时嘉 张少杰 李树文[1] 吴一民[1] Wang Peng;Zhang Debao;Zhang Yang;Zhang Haibin;Shi Jia;Zhang Shaojie;Li Shuwen;Wu Yimin(The Second Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010000,China;Inner Mongolia Autonomous Region People's Hospital,Hohhot 010000,China;First Clinical College of Medicine,Inner Mongolia Medical University,Hohhot 010000,China;Department of Anatomy(Digital Medicine Center),School of Basic Medicine,Inner Mongolia Medical University,Hohhot 010000,China)

机构地区：[1]内蒙古医科大学第二附属医院,内蒙古呼和浩特010000 [2]内蒙古自治区人民医院,内蒙古呼和浩特010000 [3]内蒙古医科大学第一临床医学院,内蒙古呼和浩特010000 [4]内蒙古医科大学基础医学院解剖学教研室(数字医学中心),内蒙古呼和浩特010000

出　　处：《中国临床解剖学杂志》2025年第1期69-75,共7页Chinese Journal of Clinical Anatomy

基　　金：内蒙古自治区自然科学基金面上项目(2022MS08019);内蒙古医科大学科研面上项目(YKD2021MS010)。

摘　　要：目的通过对不同组别的兔分别行椎板切除术,对比其各组术后3、6周TGF-β1、Smad3蛋白表达量以及两者相应的mRNA表达水平,从而探讨自体黄韧带抑制腰椎术后硬膜外纤维瘢痕形成与TGF-β1/Smad3信号通路的关系。方法将于内蒙古医科大学实验动物中心购买的48只6~8月龄的日本大白兔,分为保留黄韧带组,自体脂肪回置组,不保留黄韧带组,对不同组别进行手术造模。术后对每组间的实验动物分别饲养3、6周后进行处死(3、6周各处死8只)。采用Western blot蛋白定量分析、RTPCR技术测定各组样本中的TGF-β1、Smad3蛋白含量及mRNA表达水平,并将以上数据分别进行组间比较,分析其差异性。结果①Western blot检测结果:3、6周保留组TGF-β1、Smad3蛋白表达量小于不保留组和脂肪组(P<0.05),不保留组和脂肪组之间并未发现明显差异。②RT-PCR结果显示:对于TGF-β1、Smad3两者的mRNA表达量而言,3、6周保留组明显小于不保留组和脂肪组(P<0.05),不保留组和脂肪组之间并未见明显差异。结论腰椎手术术后,会形成不同程度的硬膜外纤维瘢痕。若在术中保留自体黄韧带,可减少硬膜外成纤维细胞的形成,其机制与TGF-β1/Smad3信号通路有关。Objective To explore the relationship between the formation of epidural fiber scar and TGF-β1/Smad3 signaling pathway after lumbar spine surgery inhibited by autologous ligamenta flava by comparing the protein expression levels of TGF-β1 and Smad3,the corresponding mRNA expression levels of TGF-β1 and Smad3 at 3 and 6 weeks after laminectomy in different groups of rabbits.Methods Forty-eight old Japanese white rabbits,aged 6 to 8 months,purchased from the Experimental Animal Center of Inner Mongolia Medical University were divided into the following group:a retention of ligamenta flava group,an autologous fat replacement group and a non-retention of ligamenta flava group.Surgical modeling was performed for different groups.After operation,the experimental animals in each group were fed for 3 weeks and 6 weeks respectively and killed(8 animals in 3 weeks and 6 weeks,respectively).The protein content and mRNA expression levels of TGF-β1 and Smad3 in samples of each group were determined by Western-blot quantitative analysis and RT-PCR,and the above data were compared between groups to analyze their differences.Results①Western-blot analysis results:The expression levels of TGF-β1 and Smad3 in the retention group were lower than those in the non-retention group and the fat group at 3 and 6 weeks(P<0.05),and there was no significant difference between the non-retention group and the fat group.②RT-PCR results showed that the mRNA expression levels of TGF-β1 and Smad3 in the retention group were significantly lower than those in the non-retention group and the fat group at 3 and 6 weeks(P<0.05),and there was no significant difference between the non-retention group and the fat group.Conclusions After lumbar surgery,different degrees of epidural fiber scar will form.Intraoperative retention of autologous ligamenta flava can reduce the formation of epidural fibroblasts by the mechanism related to the TGF-β1/Smad3 signaling pathway.

关 键 词：黄韧带 纤维瘢痕 TGF-β1/Smad3信号通路 Western blot RT-PCR技术 

分 类 号：R681.5[医药卫生—骨科学]