摘要
目的 观察含白藜芦醇的培养基培养的人眼结膜翼状胬肉(眼结膜病理性瘢痕)成纤维细胞和人胸部皮肤病理性瘢痕(皮肤病理性瘢痕)成纤维细胞增殖凋亡侵袭迁移情况,以探讨白藜芦醇对人眼结膜和皮肤病理性瘢痕成纤维细胞增殖凋亡侵袭迁移的作用.方法 采用组织块贴壁法分离原代人眼结膜和皮肤病理性瘢痕成纤维细胞.用含0(对照)、1、2、4、8、16、32、64、128、256 µg/mL白藜芦醇的培养基培养48 h时,采用CCK-8法观察人眼结膜和皮肤病理性瘢痕成纤维细胞的细胞活力(OD值).用含0、75 µg/mL白藜芦醇的培养基培养人眼结膜和皮肤病理性瘢痕成纤维细胞8 h,采用Tunel法测算细胞凋亡率.用含0、30 µg/mL白藜芦醇的培养基培养人眼结膜和皮肤病理性瘢痕成纤维细胞48 h,观察细胞的侵袭、迁移能力[侵袭率、迁移细胞数].结果 与0 µg/mL白藜芦醇比较,含128、256 µg/mL白藜芦醇的培养基培养的人眼结膜病理性瘢痕成纤维OD值均下降(P均<0.05);含64、128、256 µg/mL白藜芦醇的培养基培养的皮肤病理性瘢痕成纤维OD值均下降(P均<0.05).与0 µg/mL白藜芦醇比较,含75 µg/mL白藜芦醇的培养基培养的人眼结膜和皮肤病理性瘢痕成纤维细胞凋亡率升高(P均<0.05),含30 µg/mL白藜芦醇的培养基培养的人眼结膜和皮肤病理性瘢痕成纤维细胞侵袭率、迁移数减少(P均<0.01).结论 含>64 µg/mL白藜芦醇的培养基培养的人眼结膜和皮肤病理性瘢痕成纤维细胞活力下降,凋亡率升高,增殖、侵袭迁移能力降低.白藜芦醇可抑制人眼结膜和皮肤病理性瘢痕成纤维细胞增殖、侵袭、迁移,并促进其凋亡.
Abstract
Objective To observe the proliferation,apoptosis,invasion,and migration of human ocular conjuncti-val pterygium(ocular pathological scar)fibroblasts and human anterior thoracic skin pathological scars(skin-derived path-ological scars)fibroblasts cultured in resveratrol-containing medium,in order to investigate the effects of resveratrol on the proliferation,apoptosis,invasion,and migration of human ocular pathological scar fibroblasts and skin-derived pathologi-cal scar fibroblasts.Methods Primary human ocular pathological scar fibroblasts and skin-derived pathological scar fi-broblasts were isolated by tissue block adherent method.The cell viability(OD value)of human ocular and skin-derived pathological scar fibroblasts was detected by CCK-8 when the cells were cultured in media containing 0(control),1,2,4,8,16,32,64,128 and 256 µg/mL resveratrol for 48 h.Human ocular pathological scar fibroblasts and skin-derived pathological scar fibroblasts were cultured with 0 and 75 µg/mL resveratrol for 8 h,and the apoptosis rate was measured by Tunel method.These two kinds fibroblasts were cultured in 0 and 30 µg/mL resveratrol media for 48 h,and the migra-tion and invasion abilities of the cells(the number of invasive migratory cells)were observed.Results The OD values of human ocular pathological scar fibroblasts cultured in media containing 128 and 256 µg/mL resveratrol decreased(both P<0.05),and the OD values of skin-derived pathological scar fibroblasts cultured in media containing 64,128,and 256 µg/mL resveratrol all decreased(all P<0.05),in comparison with those at 0 µg/mL resveratrol.Compared with those at 0 µg/mL resveratrol,the apoptosis rates of human ocular pathological scar fibroblasts and skin-derived pathological scar fi-broblasts cultured in 75 µg/mL resveratrol medium increased(all P<0.05),and the number of invasion and migration cells of human ocular pathological scar fibroblasts and skin-derived pathological scar fibroblasts cultured in medium con-taining 30 µg/mL resveratrol became smaller(all P<0.01).Conclusions Human ocular pathological scar fibroblasts and skin-derived pathological scar fibroblasts cultured in resveratrol-containing medium have reduced viability,inva-sion,and migration abilities,and increased apoptosis rate.Resveratrol can significantly inhibit the proliferation,inva-sion,and migration and promote the apoptosis of human ocular pathological scar fibroblasts and skin-derived pathological scar fibroblasts.
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