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、RT-PCR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。
Abstract
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. Results ①Western-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. ② RT-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； /
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纤维瘢痕； /
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TGF-β1/Smad3信号通路； /
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Western blot； /
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RT-PCR技术
Key words
Ligamenta flava; /
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Fibrous scar; /
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TGF-β1/Smad3 signaling pathway; /
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Western-blot; /
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RT-PCR technique
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参考文献
[1] 伍敏琦, 王立, 鞠洪斌, 等. 经椎间孔椎体间融合技术治疗腰椎退行性疾病围手术期的护理[J]. 临床医学工程, 2013, 20(1):108-109. DOI: 10.3969/j.issn.1674-4659.2013.01.0108.
[2] 李树文, 杨哲, 银和平, 等. 保留黄韧带预防模型兔腰椎椎板切除后硬脊膜粘连[J].中国组织工程研究, 2019, 23(3):416-420. DOI: 10.3969/j.issn.2095-4344.0553.
[3] 李慧敏. 黄芩苷、栀子苷及其配伍通过5-LOX/LTB4通路调控小胶质细胞M1/M2极化抗脑缺血炎性损伤机制研究[D].陕西中医药大学, 2021. DOI:10.27294/d.cnki.gsxzc.2021.000008.
[4] 贾文秀, 王冬, 罗雨欣, 等. 上皮间质转化在炎症性肠病相关肠纤维化中的作用[J].胃肠病学和肝病学杂志, 2020, 29(6):695-699. DOI: 10.3969/j.issn.1006-5709.2020.06.018.
[5] 郑又菲. 植物来源神经导管搭载SHH-EMSCs移植修复大鼠坐骨神经缺损[D].江苏大学, 2021. DOI:10.27170/d.cnki.gjsuu.2021.001421.
[6] 袁兆丰, 王海平, 环大维, 等.补肾活血汤干预骨-血耦连改善大鼠激素性股骨头坏死的作用机制研究[J].中医药信息, 2023, 40(6):8-14. DOI:10.19656/j.cnki.1002-2406.20230602.
[7] 何辉, 周建辉. 罗哌卡因与利多卡因切口逐层浸润麻醉用于脊柱手术后镇痛效果比较[J].中国乡村医药, 2015, 22(7):7-8. DOI:10.19542/j.cnki.1006-5180.2015.07.003.
[8] 张帮可. 自体Lp-PRP与Lr-PRP治疗椎间盘退变的作用比较研究[D].中国人民解放军海军军医大学,2020. DOI:10.26998/d.cnki.gjuyu. 2020.000010.
[9] 陈晨羲, 张勇, 何安琪, 等.真空辅助乳腺微创旋切系统在自体脂肪移植隆胸术后乳房肿块治疗中的疗效分析[J].中国医疗美容, 2023,13(12):1-4. DOI:10.19593/j.issn.2095-0721.2023.12.001.
[10] 张德宝, 王鹏, 李琨, 等.自体黄韧带干预下兔硬膜外纤维瘢痕的形成[J].中国组织工程研究, 2025, 29(6):1168-1175. DOI：10.12307/2025.325
[11] 胡安文, 曹纬, 吴博文, 等.带蒂筋膜脂肪片修复椎管联合持续引流预防腰椎管狭窄症并硬脊膜损伤术后瘢痕粘连的效果观察[J].中国骨与关节损伤杂志, 2020, 35(1):18-21. DOI: 10.7531/j.issn.1672-9935.2020.01.005.
[12] 贾泽涛, 郑健虎, 刘国岩, 等. 脊柱结核融合后畸形继发黄韧带骨化：1例报告和综述[J]. 中国矫形外科杂志, 2024, 32(15):1385-1391. DOI:10.20184/j.cnki.Issn1005-8478.100706.
[13] 张中华, 温琦,丁娟, 等.单侧双通道内镜治疗腰椎退变疾病的研究进展[J]. 实用骨科杂志, 2023, 29(12):1100-1104. DOI:10.13795/j.cnki.sgkz.2023.12.016.
[14] 陈新焰, 陈峰, 胡建华, 等. 颈椎术后C5神经根麻痹发病机制研究进展[J].中华骨与关节外科杂志, 2023,16(12):1139-1144.
[15] 孙培锋, 辛红梅, 曲良, 等. 腰椎术后硬膜外粘连致腰椎手术失败综合征的手术治疗[J]. 实用医药杂志, 2016, 33(5):411-413.DOI:10.14172/j.issn1671-4008.2016.05.012.
[16]Xu J, Chen Y, Yue Y, et al. Reconstruction of epidural fat with engineered adipose tissue from adipose derived stem cells and PLGA in the rabbit dorsal laminectomy model[J]. Biomaterials, 2012, 33(29):6965-6973.
[17]梁雄钦, 陈锋, 范晓茜, 等.机械应力与生物因子在腰椎黄韧带肥厚过程中作用的研究进展[J].广西医学, 2021, 43(20):2481-2486. DOI: 10.11675/j.issn.0253-4304.2021.20.20.
[18]付鹏, 万小波. 细胞因子在糖尿病视网膜病变发病中作用的研究进展[J].大医生, 2023, 8(21):141-144. DOI: 10.3969/j.issn.2096-2665. 2023.21.046.
[19]谢昌楠. Perlecan对脊髓损伤后血脊髓屏障修复的作用及机制研究[D]. 南方医科大学, 2023. DOI:10.27003/d.cnki.gojyu.2023.000141.
[20]Liu P, Chen H, Yan L, et al. Laminin alpha5 modulates fibroblast proliferation in epidural fibrosis through the PI3K/AKT/mTOR signaling pathway[J]. Mol Med Rep, 2020, 21(3):1491-1500. DOI: 10.3892/mmr.2020.10967.
[21]Zhang X, Zhang J, Liu Y, et al. Pirfenidone inhibits fibroblast proliferation, migration or adhesion and reduces epidural fibrosis in rats via the PI3K/AKT signaling pathway[J]. Biochem Biophys Res Commun, 2021, 547:183-191. DOI: 10.1016/j.bbrc.2021.01.055.
[22] 李兴华, 李芳, 于瑞蒙, 等. 清热利湿化瘀方治疗湿热蕴结型盆腔炎性疾病后遗症临床研究[J]. 新中医, 2023, 55(20): 94-99. DOI:10.13457/j.cnki.jncm.2023.20.019.
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）
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