伸筋易骨法调控家兔软骨细胞增殖减轻软骨变性的实验研究

Experimental study on the effect of Shenjin Yigu method regulating the proliferation of rabbit chondrocytes to reduce the degeneration of cartilage

关键词:  伸筋易骨法  膝骨性关节炎  甲状旁腺激素相关蛋白  软骨细胞修复

Key words:  Shenjin Yigu method  knee osteoarthritis  PTHrP  chondrocyte repair

摘要:

[目的] 研究伸筋易骨法影响调控甲状旁腺激素相关蛋白（PTHrP）促进兔软骨细胞增殖减轻软骨变性的作用。[方法] 新西兰大耳白兔编号后随机分为正常组、假手术组、模型组、推拿组、玻璃酸钠组，每组各10只，改良Hulth法制备膝骨性关节炎（KOA）兔模型，推拿组采用伸筋易骨法治疗21 d，玻璃酸钠组采用关节腔玻璃酸钠注射治疗，每周1次，共治疗3周，取材后透射电镜观察细胞器情况，扫描电镜观察软骨组织表面的变性和修复情况，逆转录-聚合酶链式反应（RT-PCR）检测PTHrP mRNA的表达。[结果] RT-PCR法检测各组软骨组织PTHrP mRNA的表达，假手术组与正常组PTHrP mRNA的表达经统计学比较，无统计学意义（P>0.05），模型组与正常组比较，具有统计学意义（P<0.05），推拿组、玻璃酸钠组与模型组比较，具有统计学意义（P<0.05），推拿组与玻璃酸钠组相比，无统计学意义（P>0.05），推拿组、玻璃酸钠组与正常组相比，具有统计学意义（P<0.05）。扫描电镜检查显示，模型组软骨表面垄沟样明暗带明显紊乱，胶原纤维裸露；推拿组和玻璃酸钠组软骨表面有纤维爬过，提示软骨修复。透射电镜观察显示，模型组软骨细胞形态肥大，粗面内质网形态异常，核膜上多个膜孔，线粒体消失，胶原纤维断裂；推拿组和玻璃酸钠组软骨细胞细胞形态结构基本正常，细胞内可见多个自噬小体。[结论] 伸筋易骨法能够调控软骨合成代谢关键蛋白PTHrP的表达，减缓软骨变性，促进软骨修复，减轻KOA症状。

Abstract:

[Objective] To investigate the effect of Shenjin Yigu method on regulation of PTHrP on proliferation of rabbit chondrocytes and reduction of cartilage degeneration. [Methods] The 50 New Zealand big-eared white rabbits randomly divided into normal group,sham operation group,model group,massage group,sodium hyaluronate group,each group of 10. Improved Hulth method was use to prepare knee osteoarthritis(KOA) rabbit model,and massage group was treated with Shenjin Yigu method during 21 days. Sodium hyaluronate group was treated with articular cavity injection of sodium hyaluronate once a week,for 3 weeks,based on the transmission electron microscope after organelles,scanning electron microscopy observed on the surface of the cartilage degeneration and repair,RT-PCR detection of PTHrP mRNA expression. [Results] RT-PCR method to detect each group of cartilage PTHrP mRNA expression,and there was no significance between sham operation group and normal group in PTHrP mRNA expression (P>0.05). Compared with normal group,there was significance in model group (P<0.05). Compared with model group,the massage group,the sodium hyaluronate group had significant difference (P<0.05). Compared with the massage group,the sodium hyaluronate group had no significant (P>0.05). Compared with the normal group,the massage group,the sodium hyaluronate group had significant difference(P<0.05). Scanning electron microscope showed that the ridges and furrows on the cartilage surface of the model group were obviously disordered,and the collagen fibers were exposed. In the massage group and sodium hyaluronate group,fibers crawled over the cartilage surface,suggesting cartilage repair. Transmission electron microscopy showed that the chondrocytes in the model group were hypertrophy in morphology,abnormal morphology of rough endoplasmic reticulum,multiple membrane pores on the nuclear membrane,disappearance of mitochondria,and fracture of collagen fibers. Chondrocytes in massage group and sodium hyaluronate group had normal morphology and structure,and several autophagosomes could be seen in the cells. [Conclusion] Shenjin Yigu method can regulate the expression of PTHrP by regulating the key protein of cartilage anabolism,slow down cartilage degeneration,promote cartilage repair,and reduce KOA symptoms.