雷公藤红素对大鼠视网膜血管内皮细胞增生和凋亡的影响

Tripterine on proliferation and apoptosis of mouse retinal vascular endothelial cells

目的观察雷公藤红素对体外培养的大鼠视网膜血管内皮细胞(mouse retinal vascular endothelial cells,MRECs)增生和凋亡的影响。方法体外原代培养MRECs,通过免疫细胞化学方法检测FⅧr-Ag从而鉴定培养的MRECs;使用不同浓度的雷公藤红素(0.01mg·L-1、0.02mg·L-1、0.05mg·L-1、0.10mg·L-1、0.20mg·L-1、0.50mg·L-1)作用于原代培养的MRECs 24h、48h、72h,分别使用MTT法和流式细胞术观察其对MRECs增生和凋亡的影响。结果雷公藤红素0.02mg·L-1以上时,吸光度A值与对照组比较差异有统计学意义(P<0.05)。作用24h细胞增生抑制率分别为(8.62±4.23)%、(24.41±6.27)%、(46.51±5.95)%、(65.62±9.21)%、(72.83±8.24)%、(74.69±11.26)%;作用48h细胞增生抑制率分别为(9.34±3.78)%、(26.21±7.35)%、(51.36±6.39)%、(66.59±8.99)%、(73.19±10.24)%、(76.16±15.21)%;作用72h细胞增生抑制率分别为(11.24±4.62)%、(32.69±7.21)%、(55.28±10.25)%、(70.22±9.34)%、(75.84±13.69)%、(78.93±10.76)%。流式细胞术结果分析显示,当雷公藤红素浓度为0.05mg·L-1时,MRECs的凋亡率为(8.42±1.07)%,与空白对照组比较差异有统计学意义(P<0.05)。结论雷公藤红素对体外培养的MRECs的增生有抑制作用,并能诱导细胞的凋亡。

Objeetive To investigate the effect of tripterine on proliferation and apoptosis of mouse retinal vascular endothelial cells （MRECs）. Methods MRECs were primary cultured, and identified by the positive expression of FⅧr-Ag using immunocytochemical method. The cultured MRECs were treated with different concentrations of tripterine （0.01 mg · L^-1,0.02 mg · L^-1,0.05 mg · L ^-1,0. 10 mg · L^-1,0. 20 mg · L^-1,0.50 mg · L^-1）for 24 hours,48 hours,72 hours,the effect of tripterine on cultured MRECs proliferation was measured by MTT,the effect of tripterine on cultured MRECs apoptosis was measured by flow cytometer. Results When the concentration of tripterine was higher than 0.02 mg · L^-1, there was significant difference of A compared with control group （ P 〈0. 05 ）. MTT assay indicated that tripterine significantly suppressed proliferation of MRECs in dose-dependent and time-dependent manners. The inhibitory rate were（（9.34±3.78）%、（26.21±7.35）%、（51.36±6.39）%、（66.59±8.99）%、（73.19±10.24）%、（76.16±15.21）%for 48 hours and（11.24±4.62）%、（32.69±7.21）%、（55.28±10.25）%、（70.22±9.34）%、（75.84±13.69）%、（78.93±10.76）%for 72 hours, blow cytometry showed that the apoptosis rate of MRECs was （ 8.42 ±1.07 ） % with tripterine concentration of 0.05 mg · L^- 1, there was difference compared with control group （P 〈 0.05 ）. Conclusion Tripterine can effectively inhibit the proliferation and induce the apoptosis of MRECs.