杞精明目汤药物血浆对结膜松弛症成纤维细胞TSG-6和PTX3表达的影响  

Effects of Qijing Mingmu decoction on expression of TSG-6 and PTX3 in conjunctivochalasis

作　　者：柯梅青 张兴儒[1] 项敏泓[1] 李青松[1] 王晗敏[1] 周桂贞[1] 

机构地区：[1]上海中医药大学附属普陀医院眼科,上海200062

出　　处：《中国中医眼科杂志》2017年第2期87-92,共6页China Journal of Chinese Ophthalmology

基　　金：上海市普陀区高层次人才科研创新资助项目(2014-A-22);上海市医学重点专科(ZK2015A20)

摘　　要：目的研究杞精明目汤药物血浆对结膜松弛症成纤维细胞肿瘤坏死因子α刺激基因6(TSG-6)和穿透素3(PTX3)表达的影响,探讨结膜松弛症的发病机制及杞精明目汤对其影响作用。方法制备杞精明目汤药物血浆和无药血浆,分别用含20%、10%、5%药物血浆或无药血浆的细胞全培养基培养结膜松弛症成纤维细胞,4 h、24 h后分别提取m RNA和蛋白质,采用实时荧光定量PCR(real time PCR)技术和Western blot法检测细胞TSG-6、PTX3以及基质金属蛋白酶MMP-1、MMP-3的表达。结果 20%和10%的含药血浆均可有效上调结膜松弛症成纤维细胞TSG-6 m RNA和蛋白的表达,且作用接近(P>0.05),而5%浓度的含药血浆对TSG-6 m RNA和蛋白的表达无明显上调作用。三种浓度的含药血浆对结膜松弛症成纤维细胞PTX3 m RNA和蛋白的表达无明显作用。三种浓度的含药血浆均可明显抑制结膜松弛症成纤维细胞MMP-1 m RNA和蛋白的表达,其中20%和10%浓度对MMP-1 m RNA抑制作用较强,效果接近(P>0.05),其次为5%;20%浓度含药血浆对MMP-1蛋白表达的抑制作用最明显,10%和5%浓度对MMP-1蛋白表达的抑制作用无明显差异(P>0.05)。三种浓度的含药血浆均可有效抑制MMP-3 m RNA和蛋白的表达,其中对MMP-3 m RNA及蛋白的抑制作用最显著的均为20%浓度的含药血浆,其次为10%。结论 20%和10%浓度的杞精明目汤药物血浆能有效上调TSG-6的表达;20%、10%、5%三种浓度的含药血浆对PTX3的表达均无明显作用;三种浓度的杞精明目汤药物血浆均能有效抑制MMP-1、MMP-3的表达。OBJECTIVE To investigate the effects of Qijing Mingmu decoction on expression of TSG-6 and PTX3 in conjunctival fibroblasts of conjunctivochalasis（CCh）. METHODS Different concentration（5%, 10%, 20%）of drug serum or drug free serum were prepared. Conjunctival fibroblasts of CCh were cultured with different concen-tration of drug serum or drug free serum for 4 or 24 hours before being harvested for total RNAs or proteins, respec-tively. The expressing levels of m RNA and proteins of TSG-6, PTX3, MMP-1 and MMP-3 were detected by quantitative real time PCR（real time PCR） and western blot assay. RESULTS The 20% and 10% concentrations of drug serum containing Qijing Mingmu decoction effectively increased the expression of TSG-6 m RNA and protein, and their effects were statistically similar（P〈0.05）; while the concentration of 5% medicated serum had no significant effect on the expression of TSG-6 m RNA and protein. The concentration of 20%, 10% and 5% medicated serum had no significant effect on the expression of PTX3 m RNA and protein. Three concentrations of drug serum all significantly reduced the expression of MMP-1 m RNA and protein. The inhibition effects of 20% and 10% medicated serum onMMP-1 m RNA were obvious, but of no significant difference（P〈0.05）, and both greater than it of 5%; The most dramatic inhibition effect of medicated serum on MMP-1 protein was from 20% medicated serum, and there was no significant difference between effects of 10% and 5% concentration（P〈0.05）. All three different concentrations of drug serum signifi-cantly reduced the expression of MMP-3 m RNA and protein. The most obvious inhibitory effect of drug serum on MMP-3 m RNA and protein was from 20%, the second effective concentration was 10%. CONCLUSIONS The 20%and 10% drug serum containing Qijing Mingmu decoction could up-regulate the expression of TSG-6 in fibroblasts of conjunctivochalasis. The three concentrations of drug serum containing Qijing Mingmu decoction had no significant effect on PTX3 but t

关 键 词：结膜松弛症 杞精明目汤 肿瘤坏死因子α刺激基因6 穿透素3 

分 类 号：R777.3[医药卫生—眼科]