TGF-β与Notch信号cross-talk调控气道高反应Treg/Th17平衡机制及针刺的作用

Airway hyperreaction intends to lead bronchospasm and airway obstruction during the perioperative anesthesia, which even impact on clinical anesthesia safely. Studies have shown that T lymphocytes plays an important role in pathogenesis of airway hyperresponsiveness, but the concrete mechanism is unknown. Our previous studies demonstrated that acupuncture can imbalance Thl7/Treg during hyperreaction, mediated by expression of Notch signaling; TGF-β is the foothold of regulation of Treg/Th17, which may cooperate with Notch signal in regulating Thl7/Treg. So we hypothesized that Acupuncture can balance Treg/Th17 by regulating TGF-β and Notch signaling. To verify the above hypothesis, study on the forward analysis and backward verification combination, to confirm that TGF-β crosstalk with Notch signaling regulate Treg/Th17 balance mechanism in vitro. And on this basis, through the gene knockout technology, further analyze the mechanism of mediate the balance of Treg/Th17 by TGF-β and Notch cross-talk with airway hyperresponsiveness and the intervention of acupuncture. This task try to provide new mechanisms and experimental evidence for acupuncture to improve airway hyperresponsiveness during perioperative.

气道高反应患者易在麻醉期间发生呼吸道痉挛和气道水肿，严重影响临床麻醉安全。既往研究证实T淋巴细胞在气道高反应进程中起着重要作用，但具体机制不明。本课题组前期研究证实针刺可明显改善气道高反应性，机制与调控Notch信号介导CD4+T淋巴细胞Th17/Treg表达平衡有关；同时，TG-β也在调控Treg/Th17平衡中发挥了关键作用，可能与Notch信号协同调控Th17/Treg平衡有关。据此推测针刺改善气道高反应性与TGF-β与Notch信号的cross-talk机制维持Treg/Th17平衡有重要因果关系。为了验证上述假设，本课题拟通过动物及体外实验，正向分析和反向验证相结合证实TGF-β与Notch信号交联调控Treg/Th17平衡在气道高反应中的机制；并进一步通过基因敲除等技术探讨针刺在TGF-β与Notch交联中的作用，以期为针刺改善围术期气道高反应提供新的机制及实验依据。

气道高反应患者易在围手术期间发生呼吸道痉挛和气道水肿，严重影响患者安全。既往研究证实T淋巴细胞在气道高反应进程中起着重要作用，但具体机制不明。本课题组前期研究证实针刺可明显改善气道高反应性，机制与调控Notch信号介导CD4+T淋巴细胞Th17/Treg表达平衡有关；同时，TG-β也在调控Treg/Th17平衡中发挥了关键作用，可能与Notch信号协同调控Th17/Treg平衡有关。据此推测针刺改善气道高反应性与TGF-β与Notch信号的cross-talk机制维持Treg/Th17平衡有重要因果关系。课题通过动物及体外实验，证实TGF-β与Notch信号交联调控Treg/Th17平衡在气道高反应中的机制；并进一步通过ATG5基因敲除等技术探讨，针刺介导ATG5相关自噬介入了TGF-β与Notch交联中的作用，实验结果证实针刺对OVA致敏的气道高反应小鼠的气道高反应症状及气道炎症具有改善作用，可调节T淋巴细胞亚群稳态，降低小鼠机体炎症水平，并调节内质网应激-自噬水平。自噬基因ATG5可介导内质网应激-自噬过程，调节T淋巴细胞亚群平衡，调控气道高反应，针刺可通过ATG5基因进而通过调控内质网应激-自噬过程促进T淋巴细胞的稳态。细胞实验再次验证Notch 信号与TGF-β信号相互影响调控。麻醉药丙泊酚可明显抑制气管平滑肌的收缩，减轻气道高反应的药物，细胞研究证实其可通过自噬及钙相关通道影响小鼠成纤维细胞体外存活及增值。结合上述研究结果，进而本课题推测针刺可通过ATG5相关自噬影响细胞增值及肌细胞电活动，进而改善气道高反应性。这是本课题下一步研究的思路。为针刺改善围术期气道高反应提供新的机制及实验依据。