屈膝点按叩揉法调控L型电压依赖性钙通道影响软骨细胞代谢的机制研究

Study on the mechanism of Quxi Dian'an Kourou method regulation of L-type calcium channel affect the metabolism of chondrocytes

[目的]通过分析屈膝点按叩揉法调控L型电压依赖性钙通道影响软骨细胞合成代谢,探讨该手法防治膝骨性关节炎(KOA)的分子机制。[方法]50只新西兰大耳白兔随机分成5组,运用Hulth造模法构建KOA兔模型,治疗组采用屈膝点按叩揉法治疗,阳性对照组采用关节腔注射玻璃酸钠注射液治疗,正常组、假手术组及模型组同条件饲养。通过实时荧光定量逆转录聚合酶链反应(RT-PCR)和蛋白免疫印迹法检测L型电压依赖性钙通道α1亚基及合成代谢相关蛋白的表达。[结果]治疗组、阳性对照组Cav1.2、Cav1.3、Cav1.4、PTHr P、ColⅡ蛋白的表达均高于模型组,差异具有统计学意义(P<0.05),治疗组Cav1.2、Cav1.3、Cav1.4、PTHr P、ColⅡ蛋白的表达与阳性对照组相比无统计学差异(P>0.05)。[结论]屈膝点按叩揉法通过影响L型电压依赖性钙通道蛋白,增加细胞内钙离子浓度,上调甲状旁腺激素相关蛋白(PTHr P)的表达,进而发挥促软骨细胞合成代谢的作用。

[Objective] To explore the molecular mechanism of the manipulation prevente and cure knee osteoarthritis（KOA）, through the analysis of Quxi Dian＇an Kourou method regulates the L-type calcium channel to affect anabolism of chondrocytes. [Methods] Fifty New Zealand white rabbits were randomly divided into 5 groups, construction of KOA rabbit model by using Hulth modeling method, treatment group using the Quxi Dian＇ankou rou method, control group using the joint cavity injection of Sodium Hyaluronate Injection treatment.Normal group, sham operation group and model group were fed with the same conditions. Real-time fluorescence quantitative reverse transcription polymerase chain reaction（RT-PCR） and Western blot were used to detect the expression of L type voltage dependent Ca2 ＋-alpha 1 subunit and its synthesis metabolism related protein. [Results] The expression of Cav1.2, Cav1.3, Cav1.4, PTHr P and Col II protein in treatment group and control group were higher than that in model group, and the difference was statistically significant（ P〈0.05）. There was no significant difference in the expression of Cav1.2, Cav1.3, Cav1.4, PTHr P and Col II when the treatment group compared with the control group（P〉0.05）. [Conclusion] By Quxi Dian＇an Kourou method affecting L-type voltage dependent calcium channel protein, increase of intracellular calcium concentration, upregulate the expression of PTHr P protein and promote metabolism synthesis of cartilage cells.