基于血脊髓屏障通透性中白蛋白及星形胶质细胞深入研究龙藤通络凝胶外治大鼠骨癌痛的中枢机制

Cancer pain is one of the most common clinic symptoms associated with malignancy, it can substantial impact on patients’ activities and quality of life. About 25% newly diagnosed, 33.33% receiving threatment and 70-90% middle-advanced stage of cancer patients have different degrees of pain.How to effectively control the pain is more and more become the hot topic in the medical profession.Traditional Chinese Medicine (TCM) has been widely external used for cancer pain, it has a long history and is safe , effective, convenient, easy to expand.it can enrich the clinical treatment, but the mechanism especially central mechanism still known little, need to deepth research based on previous research. This study, based on the previous research, will Copy tibia bone cancer pain model of breast cancer in rats, the effect of Longtengtongluo(LTTL)on behavior by recording the Limbs bearing test , gait analyze and conditioned place preference analysis. Spinal cord albumin, astrocytes and IL-1β will be measured by using enzyme linked immunosorbent assay (ELISA), RT-PCR, immunohistochmistry and immunofluorescence. The specific aims of the study are to evaluate the efficacy a of LTTL gel for cancer pain and explore its central mechanism of blood-spinal cord barrier permeability in female rat models of bone cancer pain, to provide scientific experimental basis and new thought for the development of new drugs to treatment of cancer pain.

癌症疼痛是恶性肿瘤常见的临床症状，严重影响患者生活质量。约25%新诊断恶性肿瘤患者、33.33%正在接受治疗的患者以及70-90%的中、晚期癌症患者遭受不同程度的疼痛，如何有效控制癌痛越来越成为医学界关注的热点问题。中医药外用治疗癌痛具有独特的优势，其历史悠久，安全有效，方便易行，拓展了治疗癌痛的药物范围和治疗思路。本课题在原有研究基础上，复制大鼠乳腺癌胫骨骨癌痛模型，采用四足动态承重测试系统、步态分析仪、CPP条件性位置偏爱视频分析系统、免疫组化、免疫荧光、ELISA和RT-PCR等方法从疼痛行为学、脊髓背角白蛋白、星形胶质细胞和脊髓炎性介质IL-1βmRNA、蛋白质表达等不同层次深入研究龙藤通络凝胶外用对骨癌痛大鼠血脊髓屏障通透性的影响，探讨其作用的中枢机制，为临床治疗癌痛新药研发提供新思路以及实验依据。

项目背景.癌症疼痛是恶性肿瘤常见的临床症状，严重影响患者生活质量。约25%新诊断恶性肿瘤患者、33.33%正在接受治疗的患者以及70-90%的中、晚期癌症患者遭受不同程度的疼痛，如何有效控制癌痛越来越成为医学界关注的热点问题。中医药外用治疗癌痛具有独特的优势，其历史悠久，安全有效，方便易行，拓展了治疗癌痛的药物范围和治疗思路。.主要研究内容.在原有研究基础上，复制并优化大鼠Walker256乳腺癌细胞胫骨骨癌痛模型。采用机械痛觉超敏测定、热痛觉过敏检测、免疫组化、免疫荧光、ELISA和RT-PCR等方法从疼痛行为学、脊髓背角白蛋白、星形胶质细胞和脊髓炎性介质IL-1βmRNA、蛋白质表达。从不同层次，深入研究龙藤通络方外用对骨癌痛大鼠血脊髓屏障通透性的影响。.重要结果.3.5×10^5个细胞造模较佳，龙藤通络方能降低walker256胫骨癌痛大鼠模型的机械痛和热痛，增加体重，改善生活质量。造模后第7-21天，胫骨癌痛大鼠模型中随着疼痛的进展大鼠出现脊髓白蛋白和星形胶质细胞数量的增加，龙藤通络方能降低星形胶质细胞的数量，抑制IL-1βmRNA，下调IL-1β的表达，但对脊髓背角白蛋白无明显影响。.研究结论.综合以上研究内容，3.5×10^5个细胞可能是胫骨癌痛大鼠模型的更优剂量，龙藤通络方能有效治疗walker256胫骨癌痛大鼠模型的疼痛，在胫骨癌痛大鼠模型中随着疼痛的进展大鼠出现血脊髓屏障的通透性的增加。龙藤通络凝胶止痛的机制可能通过降低调星形胶质细胞的表达，抑制IL-1βmRNA，下调IL-1β的表达而降低血脊髓屏障的通透性来实现。.科学意义.本研究探讨龙藤通络方治疗骨癌痛的中枢机制，为临床治疗癌痛新药研发提供新思路以及科学实验证据，为中医理论创新提供现代生物学支撑。