针刀“调筋治骨”法治疗膝骨关节炎的效应机制及其与FAK-PI3K-AKT力学信号转导通路的相关性研究

Knee osteoarthritis (KOA), a severe disease which seriously affects the quality of life of patients, has been the focus of scholars from various countries. The effect of acupotomy for KOA has been affirmed. Our previous researches have proved that acupotomy is effective to KOA because it can prevent cartilage degradation by relieving the adhesions of connective tissues around the knee joint, but the mechanism is not clear. A number of researches have confirmed that the reason of cartilage degradation is the disorder of mechanical signal pathway because of the damage to the mechanical properties of muscle-tendon around the knee joint. Based on this, our scientific idea is that acupotomy can prevent cartilage degradation along with improvement of muscle-tendon mechanical property and restoration of knee joint’s mechanical balance through regulating the FAK-PI3K-AKT mechanical signal pathway, which is known as “tendon regulation and bone treatment”. To verify this hypothesis, we intend to adopt acupotomy to interfere KOA rabbits. In particular, our research will be focused on the muscle-tendon-cartilage mechanical property, cartilage morphology and mechanical signal pathway, by using the ultrasound elasticity imaging, nano-indentation, electron microscope and RT-PCR, etc to study the effect of acupotomy on cartilage metabolism based on regulation of the knee joint mechanical environment and mediation of the FAK-PI3K-AKT pathway. It is expected that this study will reveal the scientific basis of “tendon regulation and bone treatment” with acupotomy, so as to find the new theoretical evidence and treatment thoughts for KOA.

膝骨关节炎（KOA）是严重影响生活质量的重大疾病，已成为各国学者的关注重点。针刀治疗KOA疗效确切。我们的前期研究发现针刀可通过松解膝周软组织粘连缓解软骨退变从而治疗KOA，但起效机制尚不明确。大量研究已证实膝周肌肉-肌腱力学特性变化引起的力学信号通路转导紊乱是诱发软骨退变的重要原因。基于此，我们的科学设想是，针刀干预可调节KOA肌肉-肌腱力学特性，改善膝关节力学失衡，通过调控FAK-PI3K-AKT力学信号通路，保护软骨，即“调筋治骨”。为验证这一假说，本项目应用针刀干预KOA兔，运用超声弹性成像、纳米压痕、电镜、RT-PCR等技术，观察肌肉-肌腱力学、软骨形态学及力学信号通路相关信号分子的变化及三者的关系，探讨针刀通过调控膝关节力学环境对软骨代谢的影响以及FAK-PI3K-AKT通路在其中的介导作用，以揭示针刀通过矫正“伤筋”而“正骨”的科学性，为治疗KOA提供新的治疗思路和理论依据。

背景及目的： KOA是严重影响生活质量的重大疾病，针刀治疗通过松解膝周软组织治疗KOA疗效确切，但机制尚不明确。本项目探讨针刀干预对膝周肌肉-肌腱力学、软骨形态学及力学信号通路相关信号分子的变化及三者的关系，以揭示针刀恢复关节力学平衡，缓解软骨降解，即“调筋治骨”的科学内涵及理论基础。.方法：以改良Videman法制备KOA兔模型，行针刀、电针干预。采用改良Lequesne MG量表和膝关节被动活动度进行行为学评价；光镜、电镜观察肌肉肌腱、软骨病理表现，MRI扫描观察软骨变化；超声弹性成像对肌肉、肌腱进行力学测试；表面肌电检测肌肉运动功能；力学测试器和纳米压痕实验检测软骨力学特性；ELISA法检测血清中COMP 和尿液中CTX-Ⅱ含量； Western blot、RT-PCR检测软骨中FAK-PI3K-AKT通路蛋白、基因的表达并检测下游蛋白的水平。.结果：（1）成功制备KOA兔模型；（2）行为学方面，针刀能改善KOA兔膝关节的被动活动度，降低Lequesne MG评分；（3）形态学方面，针刀干预后光镜下软骨细胞表浅层剥脱、排列紊乱得到改善，电镜下软骨细胞胶原纤维裸露、垄沟排列消失得到减轻，肌肉细胞肌丝断裂和线粒体破坏得到改善；（4）生物力学方面，针刀可不同程度改善股四头肌肉肌腱的力学性能及肌肉疲劳程度，改善关节软骨的受力及力学性能；（5）分子生物学方面，针刀降低了软骨代谢标志物COMP和CTX-Ⅱ，上调软骨FAK-PI3K-AKT的基因和蛋白表达。（6）针刀治疗KOA兔的可能机理：①通过改善股四头肌肉肌腱的生物力学特性，恢复膝关节的力学平衡，促进膝关节稳定；②通过恢复膝关节力学平衡，激活软骨细胞FAK-PI3K-AKT信号通路，促进软骨细胞的合成代谢；③由力学信号通路介导，进一步改善软骨的受力及力学特性，减缓软骨的损伤。.结论：针刀能较好地治疗KOA，可能通过调控膝关节应力-力化学信号网络实现，部分恢复受损股四头肌肉肌腱的力学性能从而提高关节稳定性，恢复关节力学平衡，为膝关节软骨提供良性的应力刺激，由FAK-PI3K-AKT通路介导，最终调控软骨细胞的合成代谢，实现“调筋”从而“治骨”，达到治疗KOA的目的。