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目的:采用4种信号转导阻断剂研究5种天然药物姜黄素(Curcumin,Cur)、榄香烯(elemene,Ele)、三氧化二砷(arsenite trioxide,As2O3)、雷公藤内酯醇(triptolide,Tri)、莪术(rhizoma curcumae,Rc)抑制重组人碱性成纤维细胞生长因子(recombinant human basic fibroblast growth factor,rhbFGF)所诱导的牛晶状体上皮细胞(human lens epithelial cell,LEC)增殖的细胞信号转导机制。方法:将rhbFGF诱导体外培养的LEC发生增殖,用Cur、Ele、As2O3、Tri、Rc分别与其共同孵育,并用4种细胞信号转导阻断剂H7、PD98059、W7、尼卡地平(Nicardip-ine)分别作用于上述LEC。用流式细胞术(flow cytometer,FCM)检测LEC增殖细胞核抗原(proliferating cell nu-clear antigen,PCNA)的蛋白表达。结果:增殖组LEC的PCNA蛋白表达显著高于对照组(P<0.01);Cur组、Ele组、As2O3组、Tri组、Rc组的PCNA均比增殖组显著降低(P<0.01);Cur抑制LEC增殖的信号转导通路可被H7、PD98059和nicardipine阻断;Ele和As2O3均可被H7和W7阻断;Tri可被W7、nicardipine阻断;Rc可被H7、W7、PD98059和nicardipine阻断。以上均有显著性差异(P<0.01)。结论:5种天然药物Cur、Ele、As2O3、Tri、Rc抑制LEC增殖的作用是通过蛋白激酶C、丝裂原活化蛋白激酶、Ca2+-钙调蛋白、钙通道途径等信号转导机制,为寻求防治后发性白内障的有效药物提供了科学依据。
Objective: To study five natural drugs Curcumin (Cur), Elemene (Ele), Arsenite trioxide (As2O3), Triptolide (Tri) with four signal transduction inhibitors. And signal transduction of bovine lens epithelial cell (LEC) proliferation induced by recombinant human basic fibroblast growth factor (rhbFGF) induced by rhizoma curcumae(Rc) . METHODS: The proliferation of LEC cultured in vitro induced by rhbFGF was incubated with Cur, Ele, As2O3, Tri, and Rc, respectively, and four cell signal transduction agents H7, PD98059, W7 and Nicardip-ine were used. ) Acts on the above LECs, respectively. Flow cytometer (FCM) was used to detect the protein expression of LEC proliferating cell nuclear antigen (PCNA). RESULTS: The expression of PCNA protein in proliferating group LEC was significantly higher than that in control group (P<0.01). The PCNA in Cur group, Ele group, As2O3 group, Tri group and Rc group was significantly lower than that in proliferating group (P<0.01); LEC proliferation signal transduction pathway can be blocked by H7, PD98059 and nicardipine; Ele and As2O3 can be blocked by H7 and W7; Tri can be blocked by W7, niclapin; Rc can be blocked by H7, W7, PD98059 and nicardipine . There were significant differences (P<0.01). Conclusion: Curcumin, Ele, As2O3, Tri, and Rc, the five natural drugs, inhibit the proliferation of LECs by seeking signal transduction mechanisms such as protein kinase C, mitogen-activated protein kinase, Ca2+-calmodulin, and calcium channel pathways. The effective drugs for preventing and treating after-cataract provide scientific basis.