电针改善肺癌化疗后中性粒细胞减少症的神经调节机制研究

Chemotherapy-induced neutropenia, which inhibits the proliferation of hematopoietic stem/progenitor cells (HSPCs), Seriously affects the prognosis of lung cancer. It has recently been reported chemotherapy-induced sympathetic nerve injury in bone marrow (BM) impaired hematopoietic regeneration. Lines of clinic studies have demonstrated that acupuncture therapy can effectively alleviates chemotherapy-induced neutropenia and peripheral nerve injury, but the exact mechanisms are still unknown. In view of our previous investigation of sympathetic nervous system regulating hematopoiesis via pituitary adenylate cyclase activating polypeptide (PACAP) and acupuncture alleviating peripheral nerve injuries, we proposed that electroacupuncture may alleviate chemotherapy induced-neutropenia via sympathetic-hematopoietic stem/progenitor cells (HSPCs) regulatory pathway. In this study, by using chemotherapy-induced neutropenia mice models with lung cancer, treated with electroacupuncture, we will access to the effect of electroacupunture by detecting complete blood count, the population and proliferation capacity of HSPCs in the BM with FCM analysis. And then we will observe the degeneration of sympathetic nerve, the contents of several neurotrophic factors and PACAP in BM to access the neuro-protective effect of electroacupuncuture, and the action of eletroacupuncture on the secretion of PACAP. Finally, the role of PACAP in chemotherapy-induced neutropenia will be demonstrated by the administration of PACAP and its receptor antagonist into BM. In this study, we aim to disclose the potential neuro-hematopoietic regulatory context by which electroacupuncture affects chemotherapy-induced neutropenia, providing new evidences for clinical application of electroacupuncture in tumor-chemotherapy.

化疗药因抑制造血干祖细胞增殖而引起的中性粒细胞减少症严重影响肺癌预后。近期研究表明化疗药导致交感神经退变也是造血干细胞失能的重要原因。现有临床研究证实针刺可有效改善化疗后中性粒细胞减少症，但作用机制不清。本课题基于交感神经通过垂体腺苷酸环化酶激活肽（PACAP）促进造血干祖细胞增殖和电针改善外周神经损伤的前期研究，提出交感神经调控造血干祖细胞增殖介导电针改善化疗后中性粒细胞减少症的科学假说。以电针干预肺癌化疗小鼠为研究对象，采用血常规和流式细胞术观察外周血象、骨髓造血干祖细胞增殖情况，建立电针改善骨髓抑制的效应平台。采用多重免疫荧光染色观察骨髓交感神经退变，Western Blot和ELISA检测神经营养因子和PACAP含量，明确电针的神经保护效应和对PACAP分泌的影响。给予PACAP及其受体拮抗剂，明确PACAP介导针效的神经调节机制，为针刺在肺癌化疗的临床应用提供科学依据。

铂类化疗是多种肿瘤的一线治疗方法，但其使用受到严重副作用的限制，中性粒细胞减少症增加感染风险，威胁癌症患者的健康。针刺治疗肿瘤化疗后中性粒细胞减少症的安全性和有效性已经得到了既往临床试验的肯定，因此本研究试图探讨针刺作用的潜在机制。结果显示，电针足三里和三阴交能减轻顺铂诱导的中性粒细胞减少，并能增加分化多能祖细胞(MPPs)和髓系共同祖细胞(CMPs)的数量，增加正常和3LL肺癌小鼠骨髓细胞的增殖功能。进一步研究表明，电针可促进交感神经修复，增加神经生长因子、脑源性神经营养因子和垂体腺苷酸环化酶激活肽(PACAP)的产生。PACAP特异性受体PAC1的拮抗剂和激动剂分别拮抗和模拟了电针改善中性粒细胞减少症和促进骨髓造血作用。此外，骨髓基因芯片分析显示，细胞外基质可能是电针的调控靶点。因此，我们的结果提示电针可改善顺铂导致的骨髓交感神经退变，改善骨髓抑制和中性粒细胞减少，为电针防治肿瘤化疗后中性粒细胞减少症的临床应用提供了科学依据。