短刺法结合电针对兔膝骨关节炎软骨细胞增殖及凋亡的影响

付妮妮; 李学智; 杨晓光等

短刺法结合电针对兔膝骨关节炎软骨细胞增殖及凋亡的影响

Effect of Electroacupuncture with Near-to-bone Needling to Chondrocyte Apoptosis and Proliferation on the Knee Osteoarthritis Model

摘要

摘要: 目的研究短刺法结合电针对膝骨关节炎软骨细胞增殖及凋亡的影响。方法将40只新西兰兔随机分成2组，正常组（A组）10只、造模组30只。造模组动物采用Hulth-Telhag法复制实验兔左膝骨关节炎模型，6周后X线评价造模情况。将造模组再随机分为模型组(B组，n=10）、短刺组(C组，n=10）和普通刺法组(D组，n=10），第6周开始对C组和D组进行针刺后电针治疗，5 d为一个疗程，共治疗4个疗程（A、B组不行治疗）。治疗结束后X线检测造模情况，HE染色检测软骨组织病理变化情况，TUNEL检测软骨细胞凋亡情况， Western blot检测软骨细胞中天冬氨酸特异性半胱氨酸蛋白酶-3（Caspase-3）、增殖细胞核抗原（PCNA）、Bcl-2、Bax蛋白表达情况，RT-PCR技术检测软骨细胞Caspase-3、PCNA的mRNA表达。结果 HE染色结果显示：与B组相比，A组、C组与D组软骨细胞排列较整齐，关节表面光滑。 Western blot结果显示:与B组相比，C组与D组中Bcl-2、PCNA表达升高，Caspase-3与Bax表达下降；与D组相比，C组中Bcl-2、PCNA表达升高，Caspase-3与Bax表达均下降。 RT-PCR结果显示：与B组相比，C组与D组中PCNA mRNA表达升高，Caspase-3 mRNA表达下降；与D组相比，C组中PCNA mRNA表达均升高，Caspase-3 mRNA表达下降。结论短刺法结合电针可以减少软骨细胞凋亡，促进软骨细胞增殖，效果优于普通刺法结合电针。

Abstract: Objective To study the effect of electroacupuncture with near-to-bone needling on the proliferation and apoptosis of knee osteoarthritis. Methods Totally 40 New Zealand rabbits were randomly divided into two groups, the normal group (A, n =10) and the modeling group (n =30). Hulth-Telhag method was applied 30 rabbits to establish rabbit left knee osteoarthritis model, and evaluated by X-ray after 6 weeks. Then divided the 30 modeling rabbits into electroacupuncture with near-to-bone needling group (C,n =10), electroacupuncture with ordinary needling group (D,n =10) and model group (B ,n =10) randomly. Four treatments (5 d as one course of treatment) were applied to C group and D group, respectively. At the end of the experiment, modeling situation was detected by X-ray, pathological changes of cartilage was detected by HE staining ,and apoptosis of chondrocytes was checked by TUNEL staining; mRNA expressions of Caspase-3 and proliferating cell unclear antigen (PCNA) in chondrocytes were detected by RT-PCR; protein expressions of Caspase-3, PCNA, Bcl-2 and Bax in chondrocytes were detected by Western blot. Results HE stain demonstrated that the chondrocytes of A group,C group and D group were arranged in a more regular and smooth surface when compared with B group. The expressions of Bcl-2 and PCNA in C and D groups were higher than those of B group, but the expression of Caspase-3 and Bax were lower, when evaluated with Western blot；increased expressions of PCNA and Bcl-2 but decreased expressions of Caspase-3 and Bax in C group were also noticed when compared with D group. RT-PCR results demonstrated that when compared with B group, the expression of PCNA mRNA were increased but Caspase-3 mRNA were decreased in C and D groups; compared with D group, the expression of PCNA mRNA were increased but Caspase-3 mRNA were decreased in C group. Conclusion The electroacupuncture with near-to-bone needling method is superior to the electroacupuncture with ordinary needling in decreasing apoptosis and promoting proliferation of chondrocytes.

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