骨痛灵方干预αvβ3信号通路治疗肺癌骨转移疼痛的机制研究

Osseous metastasis is one of the common complications of the lung cancer. The patients with lung cancer suffer a lot from osseous metastasis. As to the pain caused by the lung cancer’s osseous metastasis, it is mainly due to the over-activity of the osteoclast which leads to the pathological changes of osteolysis. The effect of bone resorption reflects the state of the osteolysis directly. As a result, the nerve injury ,inflammatory and so on, are key to cause the pain. Integrin ɑvβ3 is thought the core molecule in the course of the osteolysis of osteoclast.. Gutongling Decoction has good clinic base and on top of that, we study the effect of this prescription suppressing the forming of the podosome to inhibit the osteolysis through the pathways of ɑvβ3/PyK2/Src/Cb1, with the rat model of the lung cancer osseous metastasis treated by the Gutongling Decoction, focusing on the osteoclast’s osteolysis, through the behavioristics, iconography, morphology and molecular biology methods. And we draw a bead on the the international problem in the field of cancer pain, to explain the possible mechanism of the treatment of tonifying the kidney and enhancing the yin, invigorating the blood and dredging the vessels to control the pain of lung cancer’s osseous metastasis. Therefore, we could show the advantage of traditional Chinese Medicine in the treatment of osseous metastasis,such as safety, efficacy, no toxic emulsion, no addiction etc. This paper is expected to provide more evidence of experiments for traditional Chinese Medicine to treat the osseous metastasis pain and to support Gutongling Decoction to be applied more widely in the clinic.

骨转移是肺癌的常见并发症之一。骨疼痛是影响患者生活质量最严重的问题。肺癌骨转移疼痛主要由于破骨细胞异常活化引起溶骨性病理改变而产生。骨吸收的作用直接反应了骨溶解的状态，骨溶解所带来的炎性反应和神经损伤等，是造成骨转移疼痛的关键。αvβ3整合素被认为是破骨细胞骨吸收过程中的核心分子。. 骨痛灵方具有良好的临床基础，我们拟在前期研究基础上，采用SD大鼠建立肺癌骨转移癌痛模型，用骨痛灵方进行治疗，以破骨细胞的骨吸收作为切入点，通过行为学、影像学、形态学和分子生物学方法，探讨其可对αvβ3/PyK2/Src/Cb1通路抑制破骨细胞足突小体形成而影响骨吸收的作用，治疗肺癌骨转移疼痛。瞄准国际肿瘤界治疗癌性疼痛的难题，阐述补肾养精，破瘀通络法控制肺癌骨转移疼痛及抑制肺癌骨转移的机制。发挥中医药治疗骨转移疼痛有效安全、无毒副反应、无成瘾性等特色及优势。为其更广泛的临床应用提供理论依据。

肺癌是临床最常见的肿瘤之一，其骨转移的发生率约30%-40%。肺癌骨转移患者常遭受着骨疼痛及病理性骨折、脊髓压迫等骨相关事件的折磨。积极治疗癌痛是WHO癌症综合规划中的四项重点之一。中医药治疗癌痛有效、经济、副作用小，极大地丰富了癌痛治疗的内容，但迫切需要继续深化其作用机制。本研究采用Lewis肺癌细胞建立C57BL/6小鼠骨癌痛动物模型，用骨痛灵方进行治疗，采用von frey测痛仪评价小鼠疼痛行为学变化；采用小动物CT扫描仪观察小鼠左下肢胫骨局部骨组织、骨密度变化；利用RT-PCR、免疫组化等方法检测骨组织标本中影响骨吸收的相关指标ανβ3、PyK2、c-Scr及Cb1的表达差异；采用激光共聚焦免疫荧光双标法检测破骨细胞中的肌动蛋白，观察足突小体的形成。从行为学、影像学、形态学和分子生物学方法等不同层面，探讨骨痛灵方治疗癌痛的机制。研究结果发现，与生理盐水组相比较，骨痛灵方治疗骨癌痛小鼠后，能够改善癌痛小鼠的体重，提高癌痛小鼠的机械痛觉缩足阈值；机制研究发现骨痛灵方能降低小鼠局部骨组织中ανβ3、PyK2、c-Scr及Cb1的mRNA及蛋白的表达水平，抑制破骨细胞足突小体形成，从而抑制破骨细胞活性，提高局部骨组织的骨密度。本研究阐述了骨痛灵方可通过改善动物行为学、局部骨密度，抑制破骨细胞足突小体形成从而影响其骨吸收来治疗肺癌骨转移疼痛，为新药研发提供科学实验依据。