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目的研究大麻素WIN55,212-2对体外培养的牛眼小梁细胞MMP-3、MMP-9及TIMP-1表达的影响,从而探讨其降眼压机制。方法采用组织块培养法对牛眼小梁细胞进行原代及传代培养,应用免疫组化方法(NSE,Ⅷ因子相关抗原染色)鉴定细胞,应用透射电镜对细胞进行形态学及生长特性观察;对传3代的小梁细胞分别施加含大麻素WIN55,212-2终浓度为0(对照组)、1、10、20、40μmol/L的培养液,48h后行MMP-3和MMP-9免疫组化SP染色,结果进行计算机图像分析并进行统计学检验。提取上清液用ELASA法检测随浓度的不同TIMP-1量的变化。结果体外培养的牛眼小梁细胞表达MMP-3及MMP-9,大麻素WIN55,212-2可促进MMP-3及MMP-9的表达,并抑制TIMP-1的表达。结论一定剂量的大麻素可以促进牛眼小梁细胞MMP-3及MMP-9的表达(P<0.05),并抑制TIMP-1的表达(P<0.01),从而达到降低眼压的目的。
Objective To investigate the effect of cannabinoid WIN55 and 212-2 on the expression of MMP-3, MMP-9 and TIMP-1 in cultured bovine trabecular meshwork cells. Methods Bovine trabecular meshwork cells were cultured in primary culture and subculture with tissue culture method. The cells were identified by immunohistochemistry (NSE, Ⅷ factor-related antigen staining), and morphological and growth characteristics were observed by transmission electron microscopy. The third generation of trabecular meshwork cells were applied with cannabinoid WIN55,212-2 final concentration of 0 (control group), 1,10,20,40μmol / L of the culture medium, 48h after the line of MMP-3 and MMP-9 immune Tissue SP staining, the results of computer image analysis and statistical tests. The supematants were extracted and the changes of the amount of different TIMP-1 were examined by ELASA. Results Bovine trabecular cells cultured in vitro expressed MMP-3 and MMP-9. Cannabinoids WIN55 and 212-2 promoted the expression of MMP-3 and MMP-9 and inhibited the expression of TIMP-1. Conclusion A certain dose of cannabinoid can promote the expression of MMP-3 and MMP-9 in bovine trabecular meshwork cells (P <0.05) and inhibit the expression of TIMP-1 (P <0.01), so as to reduce the intraocular pressure.