浅层X射线照射的间隔时间对瘢痕疙瘩成纤维细胞胶原分泌的影响  

Effects of superficial X-ray irradiation therapy at different intervals on collagen secretion of keloid fibroblasts

作　　者：郑文月 马季 臧海英 冯清蓝 王维佳 米次仁 邓成成 朱定衡 ZHENG Wenyue;MA Ji;ZANG Haiying;FENG Qinglan;WANG Weijia;MI Ciren;DENG Chengcheng;ZHU Dingheng(Southern Medical University,Guangzhou 510515,China;Dermatology Hospital,Southern Medical University,Guangzhou 510091,China)

机构地区：[1]南方医科大学,广东广州510515 [2]南方医科大学皮肤病医院,广东广州510091

出　　处：《皮肤性病诊疗学杂志》2022年第4期285-290,共6页Journal of Diagnosis and Therapy on Dermato-venereology

基　　金：广东省卫生健康委员会青年项目(A2019355);院内临床研究苗圃项目(C2019005)。

摘　　要：目的探讨不同间隔周期浅层X射线照射对人瘢痕疙瘩成纤维细胞及细胞外基质胶原蛋白表达的影响,确认最适辐照间隔时间。方法收集切除的瘢痕疙瘩组织分离培养原代瘢痕疙瘩成纤维细胞,实验组进行不同时间间隔(1、2、3、4、5、6、7 d)浅层X射线照射,总剂量均为15 Gy。采用实时定量聚合酶链反应(RT-qPCR)与Western blot检测细胞内Ⅰ、Ⅲ型胶原蛋白的相对表达,ELISA检测细胞外上清Ⅰ型胶原蛋白改变,初步验证浅层X射线治疗瘢痕疙瘩的最适辐照时间间隔。对照组与不同间隔周期浅层X射线照射组Ⅰ型、Ⅲ型胶原蛋白含量的比较采用单因素方差分析;各组间Ⅰ型、Ⅲ型胶原蛋白mRNA下降量比值行独立样本t检验,P<0.05为差异有统计学意义。结果与对照组相比,不同时间间隔浅层X射线处理后瘢痕疙瘩成纤维细胞内Ⅰ型胶原蛋白mRNA表达水平在间隔3 d(t=12.09,P<0.01)出现低峰值,Ⅲ型胶原蛋白mRNA表达水平与对照组相比差异均有统计学意义(P<0.05)。间隔3 d mRNA水平检测成纤维细胞内Ⅰ型胶原蛋白与Ⅲ型胶原蛋白的比值下降最明显,差异有统计学意义(t=6.67,P<0.01)。除间隔7 d外,各实验组HKF细胞中Ⅰ型胶原蛋白表达水平与对照组比较,差异均有统计学意义(P<0.01),在间隔1 d(t=9.97,P<0.01)出现低峰值;而HKF细胞中Ⅲ型胶原蛋白水平与对照组相比,差异均有统计学意义(P<0.01)。Ⅰ型胶原蛋白/Ⅲ型胶原蛋白比值在间隔前3 d逐渐升高,而间隔5 d(t=0.21,P>0.05)和间隔7 d(t=0.66,P>0.05)差异无统计学意义。所有实验组ELISA检测细胞外上清Ⅰ型胶原蛋白表达均下降(P<0.05)。间隔7 d照射组细胞内Ⅰ、Ⅲ型胶原蛋白及细胞外上清Ⅰ型胶原蛋白表达均出现胶原分泌回升趋势。结论照射间隔控制在3 d内能显著降低胶原蛋白的表达,对临床制定放射治疗方案具有重要意义。Objective To investigate the effect of superficial X-ray irradiation at different intervals on the expression of collagen in human keloid fibroblasts and extracellular matrix,and to confirm the optimal irradiation interval.Methods The excised keloid tissue was collected and cultured primary human keloid fibroblasts.The third passage cells were used to set up an experimental group and a control group.Intervals(1,2,3,4,5,6,7 days)with superficial X-rays.Real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot were used to detect the relative expression changes of collagen typeⅠandⅢin cells.ELISA was used to detect the changes of typeⅠcollagen in the extracellular supernatant,and preliminarily verified the optimal irradiation time interval for superficial X-ray therapy of keloids.One-way ANOVA was used to compare superficial X-ray irradiation groups at different intervals with the control group.The proportion of collagen typeⅠand typeⅢcollagen mRNA decrease in each group was tested by independent samples t test.Results Compared with the control group,the expression level of typeⅠcollagen mRNA in keloid fibroblasts in experimental group after shallow X-ray treatment at different time intervals showed a low peak at an interval of 3 days(t=12.09,P<0.01).The expression level of typeⅢcollagen mRNA in experimental group had statistical significance decrease compared with the control group(P<0.05).The ratio of typeⅠcollagen to typeⅢcollagen in fibroblasts decreased most significantly at an interval of 3 days(t=6.67,P<0.01).The expression level of typeⅠcollagen in HKF cells of each experimental group was significantly different from that of the control group(except for the interval of 7 days)(P<0.01),and there was a low peak at the interval of 1 day(t=9.97,P<0.01);The level of typeⅢcollagen in HKF cells in experimental group was significantly different from that in the control group(P<0.01).The ratio of typeⅠcollagen to typeⅢcollagen increased gradually in the first 3 days,but ther

关 键 词：瘢痕疙瘩 浅层X射线 胶原 成纤维细胞 

分 类 号：R622[医药卫生—整形外科]