目的:探讨垂体腺苷酸环化酶激活多肽 (PACAP) 的衍生物小环肽C*HSDGIC*(CHC)对中波紫外线(UVB)诱导的人视网膜色素上皮(RPE)细胞凋亡的影响.方法:Western blotting检测人RPE细胞PACAP 1型受体(PAC1受体)表达;RPE细胞接受0.2 J/cm2 UVB照射,随后实验组给予含不同浓度CHC的无血清培养基培养,对照组给予无血清培养基培养;CCK-8法检测CHC对RPE细胞活性的影响;流式细胞术Annexin-V/PI双染检测RPE细胞凋亡情况,JC-1染色检测线粒体膜电位.结果:Western blotting显示RPE细胞表达PAC1受体,CCK-8法显示CHC促细胞活性的最佳浓度为100 μmol/L,细胞活性比对照组增加(34.23±3.39)%(P<0.05),抗凋亡最佳浓度也为100 μmol/L,细胞活性比对照组增加(20.10±1.48)%(P<0.05).流式细胞仪分析显示,100 μmol/L CHC能使UVB照射后凋亡细胞比例下降 (5.63±1.49)%,线粒体去极化细胞比例下降(5.2±0.5)% (P<0.05).结论:人RPE细胞存在PAC1受体,小环肽C*HSDGIC*对人RPE细胞有促进活性和拮抗凋亡的作用.%AIM: To investigale the influence of C* HSDGIC* (CHC) , a cyclopeptide from the cyclizalion of [ pituitary adenylate cyclase - aclivaling polypeptide 1 - 5, PACAP (1 - 5 ) ] with disulfide, on the proliferation and apop-Losis of cultured human relinal pigmenl epilhelial (RPE) cells induced by ullraviolel B(UVB). METHODS; The expression of PACAP Lype 1 (PAC1) receptor in human RPE cells was identified by Weslern blotting. The cells were exposed Lo UVB irradialion and cultured in fresh medium wilh or withoul gradient concentrations (1 nmol/L Lo 1 mmol/L) of CHC. The viability of the cells was determined by CCK - 8 assay. The early apoplosis of the cells was detected by flow cylometry wilh annexin V and propidium iodide staining. The milochondrial menbrane potenlial was delected by flow cytomelry with JC - 1 slaining. RESULTS: The PAC1 receptor in human RPE cells was idenlified by Weslern blolling. The best results of CHC on the proliferation and anti - apoptosis of human RPE cells were achieved at the concentration of 100 μmol/L, which increased the viability by ( 34. 23 ± 3. 39) % and ( 20. 10 ± 1. 48 ) % , respectively. The percentage of apoptolic cells was decreased by (5. 63 ± 1.49) % with CHC treatmenl ( 100 μmol/L) after UVB irradiation,and the percentage of milochon-drium - depolarizing cells was decreased by (5.2 ±0. 5) % . CONCLUSION: PAC1 receptor exisls in human RPE cells. C * HSDGIC * increases the viability of RPE cells and atlenuates UVB - induced apoplosis.
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