破血消癥法对hRPE细胞增殖凋亡和MAPK、PI3K/Akt信号通路表达的影响

Effects of the method of removing blood stasis and resolving masses on human RPE cells proliferation and apoptosis and expression of MAPK and PI3K/Akt signaling pathway.The proliferation of retinal fibrous membrane is a common pathological change of a variety of ocular fundus diseases, and is also the fundamental reason for the failure of surgical treatment of fundus diseases. At present, there is lack of effective method to control the formation of proliferative membrane. The disease belongs to Chinese medicine"mass accumulation" category. Over the past 10 years, our preliminary study of Sanxuemingmu Tablet on the main treatment of retinal hemorrhage showed the drug could promote the absorption of fundus hemorrhage, but also inhibited the formation and development of PVR. But this medicine had main function of activating blood circulation and alleviating liquid retention and function of softening and resolving hard mass would be strengthened. Based on this, the research group put forward to using prescription of removing blood stasis and resolving masses to intervene the retinal fiber membrane hyperplasia. Through in vitro and in vivo experiments the effects of Chinese herbal compound of removing blood stasis and resolving masses on the RPE cells proliferation and apoptosis and expression of MAPK and PI3K/Akt signaling pathway, and controlling effects of Chinese herbal compound of removing blood stasis and resolving masses on inhibition of PVR and MAPK and PI3K/Akt signaling pathway of rabbits’eyes will be observed, which will confirm effects of that method of removing blood stasis and resolving masses on retinal fiber hyperplasia disease and the related signaling pathway expression.

视网膜纤维膜增生是多种眼底疾病共同的病理改变，也是眼底疾病手术治疗失败的根本原因，目前尚缺乏能有效控制增殖膜形成的治疗方法。该病当属中医“症瘕积聚”范畴。近10多年来，我们对主要治疗眼底出血的散血明目片的前期研究表明，该药在促进眼底出血吸收的同时，也有较好抑制PVR的形成和发展的作用，但该药以活血利水为主，软坚散结作用尚待增强。基于此，本课题组有针对性的提出对视网膜纤维膜增生病变，采用破血消癥法加以组方干预。拟从体外和体内实验两方面着手，分别观察破血消癥中药复方含药血清对hRPE细胞增殖凋亡和对MAPK、PI3K/AKT信号通路表达的影响、破血消癥中药复方抑制兔眼PVR和对MAPK及PI3K/Akt信号通路表达的调控作用，亦即明确破血消癥法对视网膜纤维膜增生性疾病的具体作用及对相关信号通路表达的影响。

视网膜纤维膜增生是多种眼底疾病共同的病理改变，也是眼底疾病手术治疗失败的根本原因，目前尚缺乏能有效控制增殖膜形成的治疗方法。该病当属中医“症瘕积聚”范畴。近10多年来，课题组对主要治疗眼底出血的散血明目片的前期研究表明，该药在促进眼底出血吸收的同时，也有较好抑制PVR的形成和发展的作用，但该药以活血利水为主，软坚散结作用尚待增强。基于此，本课题组有针对性的提出对视网膜纤维膜增生病变，采用破血消癥法加以组方干预。拟从体外和体内实验两方面着手，分别观察破血消癥中药复方含药血清对hRPE细胞增殖凋亡和对MAPK、PI3K/AKT信号通路表达的影响、破血消癥中药复方抑制兔眼PVR和对MAPK及PI3K/Akt信号通路表达的调控作用，亦即明确破血消癥法对视网膜纤维膜增生性疾病的具体作用及对相关信号通路表达的影响。.研究通过体外培养 RPE 细胞，观察破血消癥中药复方对 RPE 细胞增殖与凋亡的影响。结果显示破血消癥中药复方可能通过调控 p38MAPK、ERK、PI3K 信号通路有效上调Caspase-3 蛋白表达，下调 CyclinD1、p38MAPK、ERK、PI3K 蛋白表达，抑制细胞迁移，并诱导细胞凋亡。又通过建立兔外伤性增殖性玻璃体视网膜病变模型，模拟眼底视网膜纤维膜增生改变，观察破血消癥中药复方对兔外伤性增殖性玻璃体视网膜病变的防治作用，并探究其作用机制。结果显示，破血消癥中药复方可能调控 p38MAPK、PI3K、ERK 通路有效上调外伤性 PVR兔视网膜增殖膜 Caspase-3 蛋白表达、下调 CyclinD1、p38MAPK、ERK、PI3K 蛋白的表达，抑制细胞增殖、促进细胞凋亡，从而改善外伤性 PVR 兔视网膜形态结构，减少炎性细胞浸润、减轻细胞水肿、预防增生组织的产生，有效防治外伤性 PVR。体外实验与体内实验结果相一致，由此得出结论，破血消癥中药复方可能通过抑制细胞增殖、诱导细胞凋亡来控制视网膜纤维膜增生的发生与发展，由此维持视网膜正常的形态结构，阻止严重并发症的发生。.破血消癥中药复方组成药物来源广泛，毒副作用小，安全性大。本实验的研究将为视网膜纤维膜增生性疾病的中医药防治提供新方法、新思路，为临床运用提供新前景。