芍药苷对IDD兔Th1/Th2平衡漂移的影响及其机制的研究

The nucleus pulposus of the intervertebral disc is isolated from the immune system of the host by the anulus fibrosus and the cartilaginous endplates. Once the nucleus pulposus is exposed to the host's immune system, an autoimmune reaction is evoked. The imflamation reaction is the pathological mechanism of discogenic low back pain and intervertebral disc degeneration (IDD) is its initiating agent. In our previous research, the anular puncture model of IDD was established in rabbits by Sobajima's anular puncture method,and a higher degree of TNF-α expression was found in discs after tear of anular fibers. So a condition where Th1 response is supposed to be augmented during the proceed of IDD. Deterioration of Th1/Th2 balance was found in several autoimmunity-related diseases,and T-bet and GATA-3 were the key point to influence the direction of the Th1/Th2 polarization. Paeonia were contained in lots of traditional Chinese medicinal formula in the treatment of low back pain. Peoniflorin is one of the major bioactive components of paeonia,and has reportedly exhibited antiinflammatory effect, analgesic, modulating immunization and so on. The present study will carry out to investigate the effects of paeoniflorin in an experimental model of IDD in rabbits.Aim to found if paeoniflorin could modulating ratio of GATA-3/T-bet, regulating Th1/Th2 polarization, relieving imflamation reaction by RT-PCR，ELISA，Westen-blot and immunohistochemistry. The research hope to find a new target for IDD treatment,and explore a new Chinese herbal for low back pain.

正常髓核与免疫系统隔绝，当纤维环破损后，髓核就暴露于免疫系统中，激发自身免疫炎性反应，是椎间盘源性腰痛的病理基础。椎间盘退变(IDD)是盘源性腰痛发生的始动因素。在前期研究中，我们参照Sobajima的方法成功构建了兔IDD模型，发现纤维环破裂会上调TNF-α等炎性因子的水平，由此推测IDD的发生可能与Th1优势状态有关。Th1/Th2平衡漂移见于多种自身免疫性疾病，而T-bet和GATA-3是影响Th1/Th2极化的关键因素。诸多经典治疗腰痛的方剂中均含有芍药。芍药中的主要活性单体芍药苷具有抗炎、镇痛、调节免疫等作用。本项目拟采用RT-PCR，ELISA，Westen-blot及免疫组化等方法，旨在揭示芍药苷在IDD发生的病程中，可以调节GATA-3/T-bet比例，逆转Th1/Th2漂移状态，减轻炎性反应的程度，延缓椎间盘退变。有望为盘源性腰痛的治疗寻找新的靶点，为开发中药新药提供依据

正常髓核与免疫系统隔绝，当纤维环破损后，髓核就暴露于免疫系统中，激发自身免疫炎性反应，是椎间盘源性腰痛的病理基础。椎间盘退变(IDD)是盘源性腰痛发生的始动因素。在前期研究中，我们参照Sobajima的方法成功构建了兔IDD模型，发现纤维环破裂会上调TNF-α等炎性因子的水平，由此推测IDD的发生可能与Th1优势状态有关。Th1/Th2平衡漂移见于多种自身免疫性疾病，而T-bet和GATA-3是影响Th1/Th2极化的关键因素。诸多经典治疗腰痛的方剂中均含有芍药。芍药中的主要活性单体芍药苷具有抗炎、镇痛、调节免疫等作用。方法：本项目采用RT-PCR，ELISA，Westen-blot及免疫组化等方法，揭示芍药苷在IDD发生的病程中的作用。实验结果：与术前相比，造模后兔血清及椎间盘中TNFα与IFN-γ含量明显增高，椎间盘中T-bet含量明显增高；给予芍药苷干预后，兔血清及椎间盘中TNFα与IFN-γ含量明显降低，椎间盘中T-bet含量明显降低。结论：生理屏障损伤后椎间盘发生退变的机制与Th1优势状态有关，芍药苷可以调节GATA-3/T-bet比例，逆转Th1/Th2漂移状态，减轻炎性反应的程度，延缓椎间盘退变。意义：有望为盘源性腰痛的治疗寻找新的靶点，为开发中药新药提供依据