摘要
目的检测内皮素(endothelin-1,ET-1)刺激人视网膜色素上皮(retinal pigment epithelial,RPE)细胞后ET-1的自分泌。方法用体外培养的RPE细胞,经10-11mol.L-1、10-9mol.L-1、10-7mol.L-1ET-1作用后,用免疫组织化学和原位杂交检测RPE细胞ET-1的表达和一氧化氮合酶1(nitric oxide synthase1,NOS1)的表达,并用LeicaQ750图像分析仪定量分析表达量;用放射免疫测定检测细胞调理液中ET-1和6-酮-前列腺素F1α(6-Keto-PGF1α)含量。结果在未经ET-1作用的RPE中ET-1、ET-1 mRNA和NOS1仅见少量细胞微弱表达,3个浓度ET-1作用24h后,图像分析在不同浓度ET-1刺激后均检测出ET-1的表达增强,并有浓度依赖性(F=9.511,P<0.01),NOS1的表达也增强(F=16.113,P<0.01),两者有直线相关性(r=0.685,P<0.05)。在调理液的检测中,ET-1和6-Keto-PGF1α的含量均随着ET-1作用浓度的增加而增加,各作用浓度两者含量均有统计学差异(前者F=23.67,P<0.01;后者F=44.431,P<0.01);并且两者有直线相关性(r=0.85,P<0.01)。结论ET-1刺激RPE细胞表达并分泌ET-1,同时伴随的NOS1表达增加和6-Keto-PGF1α分泌增加可能是针对ET-1的负向调节作用。
[ Abstract ] Objective To investigate the autocrine of endothelin-1 （ ET-1 ） in human retinal pigment epithelial （hRPE） cells after stimulated by ET-1. Methocls The hRRE cells were ctdtttred and sttmtdatedby 10^-11 mol. L^-1、10^-9 mol· L^-1 and 10^-7 mol · L^-1 of ET-1 ,the expression of ET-1 and nitric oxide synthase 1 （NOS1） was determined by tmmunocytoehemistry and hybridization in situ, and the expressive value was quantitative analyzed by LeieaQ750 image analysator. The content of ET-1 and 6-Keto-PGF1α was quantified by radiotmmunoasaay in supernatant fluid. Results The expression of NOS1, ET-1 and ET-1 mRNA was weakly expressed in the RPE cells without treated by ET-1 ,and the expression of ET-1 and ET-1 mRNA was gradlmlly enhanced after treated by 10^-11 mol· L^-1 ,10^-9 mol· L^-1 and 10^-7 mol· L^-1 ET-1 for 24 hours（F=9.511 ,P〈0.01 ）,and that of NOS1 also enhanced（F= 16.113 ,P 〈0.01 ） ;And there were linear correla- tion in the expression of ET-1 mRNA and NOS1 （ r =0.685 ,P 〈0.05 ） after quanlifled by image Analysis. ET-1 caused a significant increase in levels compared with the control without treated by ET-1. There was a concentration-dependent significantly increase of secretion in ET-1 and 6-Keto-PGF1α in hRPE cells in response to 10^-11 mol· L^-1 ,10^-9 mol · L^-1 and 10^-7 mol · L^-1 ET-1 （F=23.67,P〈 0. 01 ;F=44.431 ,P 〈0.01, respectively） ,and there were linear correlation in the oncentration of ET-1 and 6-Keto-PGF1α （ r = 0 . 85, P 〈 0.01 ） in supernatant luid. Conclusion The expression and release of ET-1 in hRPE cells is positive regulated by ET-1 differentially, the expression of NOS1 and secretiotx of 6-KetoPGF1α y be related to negative regulation of ET-1. [ Rec Adv Ophthalmol 2007 ;27 （3） ： 161-164 ]
出处
《眼科新进展》
CAS
2007年第3期161-164,共4页
Recent Advances in Ophthalmology
基金
国家自然科学基金资助(编号:39970780)~~
