单侧与双侧翼状胬肉成纤维细胞生物学行为的差异及紫外线对其的影响  

Difference in cell biological behavior and ultraviolet radiation effect between double-head and unilateral pterygium fibroblasts cultured in vitro

作　　者：彭娟[1] 毛雁[1] 姚达强 董舒婷 沙翔垠[1] Peng Juan;Mao Yan;Yao Daqiang;Dong Shuting;Sha Xiangyin(Department of Ophthalmology,the Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,China)

机构地区：[1]广州医科大学附属第二医院眼科,510260

出　　处：《中华实验眼科杂志》2020年第5期388-394,共7页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目(81100649);广东高校优秀青年创新人才培育项目(LYM10112)。

摘　　要：目的探讨单侧与双侧人翼状胬肉成纤维细胞(HPFs)增生和移行的差异及紫外线对不同部位成纤维细胞增生和移行的影响。方法收集2017年3月至2018年3月于广州医科大学附属第二医院行翼状胬肉切除+自体结膜瓣转位术的双侧及单侧翼状胬肉患者各19例19眼的翼状胬肉组织。6例12眼正常结膜组织来源于同期供体眼球。根据病种和培养取材部位分为鼻侧HPFs(HPFs-N)19例、颞侧HPFs(HPFs-T)19例、单侧HPFs 19例和人正常结膜成纤维细胞(HCFs)12例,分别取自双侧翼状胬肉的鼻侧、双侧翼状胬肉的颞侧、单侧翼状胬肉和正常结膜。将以上成纤维细胞分为紫外线照射组和普通光照组,分别接受紫外线B(UVB)和普通光照照射。采用MTT法检测细胞生长曲线,细胞划痕试验检测48 h细胞划痕愈合率,免疫荧光法检测α平滑肌肌动蛋白(α-SMA)的表达。比较各成纤维细胞增生和移行能力的差异。比较紫外线照射组和普通光照组细胞划痕愈合率、生长曲线及α-SMA表达的改变。结果组织块培养法可见HCFs及HPFs呈梭形生长。各成纤维细胞体外培养1~7 d生长速度逐渐加快,HPFs-N生长最快,HCFs生长速度最慢。在紫外线照射下各成纤维细胞生长速度较普通光照组明显加快。4种成纤维细胞不同光照条件下细胞划痕愈合率、α-SMA阳性细胞表达率比较差异均有统计学意义(F组别=158.064,P<0.05;F细胞类型=326.582,P<0.05.F组别=4.731,P<0.05;F细胞类型=172.813,P<0.05),其中普通光照条件下,48 h后HPFs-N的细胞划痕愈合率为(79.67±0.86)%,明显高于HPFs-T、单侧HPFs和HCFs的(54.04±0.33)%、(64.12±0.21)%和(58.86±0.41)%,HPFs-N的α-SMA阳性细胞表达率为(28.87±1.02)%,明显高于HPFs-T、单侧HPFs和HCFs的(13.67±0.23)%、(20.35±1.72)%和(5.12±0.45)%,差异均有统计学意义(均P<0.05);与普通光照组比较,紫外线照射组各成纤维细胞的细胞划痕愈合率、α-SMA阳性细胞表达率明�Objective To explore the difference in the proliferation,migration and ultraviolet radiation effect between double-head and unilateral pterygium fibroblasts(HPFs)cultured in vitro.Methods Pterygium tissue was obtained from patients who underwent pterygium excision with conjunctival transposition from March 2017 to March 2018 in the Second Affiliated Hospital of Guangzhou Medical University.Nineteen cases with bilateral pterygium and 19 cases with unilateral pterygium were selected for this research.Twelve cases of normal conjunctival tissue were obtained from donor eyes.The fibroblasts were divided into HPFs-nasal(HPFs-N),HPFs-temporal(HPFs-T),unilateral HPFs and human conjunctiva fibroblasts(HCFs),which was taken from the nasal side of the bilateral pterygium,the temporal side of the bilateral pterygium,the unilateral pterygium and the normal conjunctiva,respectively.Human pterygium and normal conjunctival fibroblasts were isolated and cultured by tissue culture method.The fibroblasts were divided into an ultraviolet irradiation group and a normal light illumination group.The cell growth curve was detected by methyl thiazolyl tetrazolium(MTT)assay.The cell scratch healing rate was detected by the cell scratch test after 48 hours.The expression of Symbola-smooth muscle actin(Symbola-SMA)was detected by immunofluorescence staining,and the number of positive cells in each group was compared.The fibroblasts cultured in vitro were irradiated with ultraviolet irradiation,and the cell scratch healing rate,growth curve and expression of Symbola-SMA were observed.The study protocol was approved by the Ethics Committee of the Second Affiliated Hospital of Guangzhou Medical University.Written informed consent was obtained from each subject.Results The pterygium and conjunctival fibroblasts were spindle shaped,and the growth rate was gradually increased from day 1 to day 7.The growth rate of HPFs-N was the fastest,and the growth rate of HCFs was the slowest.The growth rate of the four types of fibroblasts was significantly

关 键 词：翼状胬肉 成纤维细胞 细胞培养 增生 移行 紫外线 

分 类 号：R77[医药卫生—眼科]