人视网膜色素上皮细胞增殖与川芎嗪的影响

Effects of Tetramethylpyrazine on the proliferation of human retinal pigment epithelial cells

目的:观察糖基化终产物(advanced glycation end products,AGEs)对人视网膜色素上皮细胞增殖的影响,以及川芎嗪对AGEs诱导的人视网膜色素上皮细胞增殖的拮抗作用。方法:实验于2003-09/2004-07在中南大学湘雅二医院中心实验室完成。①实验分组:原代培养人视网膜色素上皮细胞,第3￣8代用于实验,设空白对照组,DMEM培养基中不加入任何药物;AGEs对照物组,培养液中含100mg/L的AGEs对照物;AGEs组,培养液中含10,50,100,200,400mg/L的AGEs,分别干预视网膜色素上皮细胞24h及48h。②实验操作:川芎嗪干预实验:也设空白对照组;AGEs对照物组;AGEs组,培养液中含100mg/L的AGEs;AGEs+川芎嗪组,培养液中加入10,20,40,80,160,320,640mg/L川芎嗪30min后加入100mg/L的AGEs,分别干预视网膜色素上皮细胞24h。③实验评估:MTT微量酶比色法检测视网膜色素上皮细胞增殖情况。结果:①视网膜色素上皮细胞增殖情况:不同浓度AGEs组细胞增殖率均高于对照物组(P<0.01),100mg/LAGEs组增殖率(24,48h分别为39.45%,31.97%)高于其他组(P<0.01),48h与24h结果一致。②川芎嗪的抑制效应:与AGEs组比较,10,20,40,80,160mg/L川芎嗪对AGEs诱导的视网膜色素上皮细胞增殖无抑制作用(P>0.05),320及640mg/L川芎嗪有抑制作用,其抑制率为8.96%,15.08%(P<0.01)。结论:①AGEs可诱导人视网膜色素上皮细胞增殖。②低浓度川芎嗪对AGEs诱导的人视网膜色素上皮细胞增殖无抑制作用,高浓度川芎嗪具有抑制作用。

AIM： To investigate the effect of advanced glycation end products （AGEs） on the proliferation of human retinal pigment epithelial （RPE） cells, and the inhibitory effect of Tetramethylpyrazine （TMP）. METHODS： The experiment was conducted at the Central Laboratory of the Second Xiangya Hospital, Central South University from September 2003 to July 2004. ①The third to eighth primarily cultured human RPE cells ware selected and those cultured in DMEM medium without any drug as blank control, in culture solution containing 100 mg/L AGEs control as the AGEs control group, and in solution containing 10, 50, 100, 200, and 400 mg/L AGEs for 24 and 48 hours as the AGEs groups. ②Meanwhile, in the TMP intervention test, besides the three previous groups, there ware AGEs plus TMP group, in which the RPE cells ware firstly exposed to TMP of 10, 20, 40, 80, 160, 320 and 640 mg/L for 30 minutes, following by 100 mg/L AGEs for 24 hours.③3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide （MTT） assay was adopted to detect the proliferation of RPE cells. RESULTS： ①The proliferative rate of RPE cells was stimulated by AGEs at different concentrations, and higher than that of the AGEs control group （P 〈 0.01）, in particular, the 100 mg/L group was the highest in 24 hours and 48 hours, respectively （39.45%, 31.97%, P 〈 0.01）. ②Compared with the AGEs groups, TMP at concentrations of 10, 20, 40, 80, and 160 mg/L did not inhibit the proliferation of RPE cells induced by AGEs （P 〉 0.05）, while at concentrations of 320 and 640 mg/L could inhibit the growth （8.96%, 15.08%, P 〈 0.01 ）. CONCLUSION： ①AGEs promote the proliferation of RPE cells.②Low doses TMP could not inhibit the proliferation of RPE cells induced by AGEs, while high doses TMP has the inhibitory effects.