基于α7nAChR介导的NF-κB和JAK2/STAT3信号通路的针刺干预干眼炎症的作用机制研究

Dry eye is a common ocular surface disease, often accompanied by local inflammation. Clinical practice have proved that acupuncture effectively treat dry eye with inflammation. Our previous study found that acupuncture could regulate the cholinergic pathway and alleviate dry eye with inflammation. This project will further explore the effects and mechanisms of acupuncture on α7nAChR and its mediated NF-κB and JAK2/STAT3 signaling pathways and inflammation. The rabbit models of dry eye were treated with acupuncture according to the effective acupoint in clinical treatment, and then analysis the contents of ACh and α7nAChR in lacrimal gland, cornea and conjunctiva; assess expression of NF-κB, JAK2, STAT3 genes and proteins which are associated with cholinergic anti-inflammatory pathways at the molecular and protein levels, respectively. Moreover, this project will explore the change of the related-inflammatory factors IL - 1, IL-6,TNF-α and so on in tissues and tears. More importantly, this project will clarify the effect and mechanism of acupuncture on the change of NF-κB, JAK2 and STAT3 of the main signal transduction pathways mediated by α7nAChR in rabbits with dry eye, subsequently to inhibit the release of inflammatory cytokines, and reduce the ocular surface inflammation using α7nAChR antagonist to block the cholinergic anti-inflammatory pathway. This project will explicit the effect and mechanism of acupuncture on the regulation of ocular inflammation by cholinergic anti-inflammatory pathway. It is hoped that this study will make a breakthrough in the mechanism of acupuncture for dry eye inflammation.

干眼是临床上常见眼表疾病，常伴有局部炎症，临床证实针刺有效治疗干眼炎症，本课题组前期工作发现针刺可调节胆碱能通路减轻干眼炎症。本项目将进一步探讨针刺调节α7nAChR及其介导的NF-κB和JAK2/STAT信号通路，抑制炎症的效应及机制。采用临床有效穴位对干眼模型兔进行针刺治疗，从分子及蛋白水平各有侧重分析针刺对泪腺、角膜及结膜组织ACh、α7nAChR水平；胆碱能抗炎通路关键基因及蛋白如NF-κB、JAK2、STAT3的表达；组织及泪液中相关细胞炎症因子IL-1、IL-6 和TNF-α等的变化。并用α7nAChR拮抗剂阻断胆碱能抗炎通路，阐述针刺调节干眼炎症模型兔中α7nAChR及其介导的信号转导通路NF-κB和JAK2/STAT3表达，抑制炎症因子释放，从而减轻眼表炎症的机制。该工作将明确针刺调控胆碱能抗炎通路干预干眼炎症的作用机制。有望通过该项研究对针刺治疗干眼炎症的机制产生突破性进

干眼是临床上常见眼表疾病，常伴有局部炎症，临床证实针刺有效治疗干眼炎症，本课题组前期工作发现针刺可调节胆碱能通路减轻干眼炎症。本项目进一步探讨针刺调节α7nAChR受体及其介导的NF-κB和JAK2/STAT信号通路，抑制炎症的效应及机制。采用临床有效穴位对干眼模型兔进行针刺治疗，从分子及蛋白水平各有侧重分析针刺对泪腺、角膜及结膜组织ACh、α7nAChR水平；胆碱能抗炎通路关键基因及蛋白如NF-κB、JAK2、STAT3等表达；组织及泪液中相关细胞炎症因子IL-1、IL-6和TNF-α变化。并用α7nAChR拮抗剂阻断胆碱能抗炎通路，阐述针刺调节干眼模型兔中α7nAChR及其介导的信号转导通路NF-κB和JAK2/STAT3表达，抑制炎症因子释放，从而减轻眼表炎症的机制。研究结果表明：①针刺可以抑制干眼眼表炎症反应：减少角膜荧光染色，增加泪液分泌和泪膜破裂时间，降低泪液渗透压；抑制异常活化的角膜基质层细胞，改善角膜神经异常形态；减少角膜上皮脱落和淋巴细胞浸润，改善角膜超微结构；②针刺抗炎效应与胆碱能抗炎通路有关：针刺可以增加Ach和α7nAChR受体的表达，下调JAK2/STAT3/NF-KB蛋白和相关促炎细胞因子、趋化因子的表达水平，这些有益作用与α7nAChR的上调以及JAK2/STAT3/NF-KB的下调有关。③进一步证实了针刺对干眼炎症胆碱能抗炎通路的调控机制：使用α7nAChR拮抗剂后，针刺的有益作用受到抑制，这表明针刺对干眼的治疗是通过α7nAChR介导的抗炎途径，即针刺可以通过调控α7nAChR抑制JAK2/STAT3/NFKB信号通路激活并且降低炎症因子的表达来起到抗炎作用。所以α7nAChR介导的JAK2/STAT3/NF-KB信号通路可能充当针刺治疗干眼的潜在治疗靶点。