摘要
目的研究碱性调宁蛋白(CNN1)在系统性硬化症(SSc)中的表达及其对成纤维细胞的作用。方法收集SSc患者皮肤活检组织19例,正常人的皮肤活检组织21例。通过Real-time PCR方法检测皮肤组织中CNN1和α-SMA的基因表达;通过免疫组化方法检测皮肤组织中CNN1的蛋白表达;利用RNA干扰方法降低成纤维细胞中CNN1的基因表达,通过Real-time PCR检测细胞中CNN1及纤维化相关基因α-SMA、CTGF、COL1A1、COL1A2、COL3A1的mRNA相对表达量;应用细胞实时增殖检测系统(xCELLigence系统)检测细胞增殖。结果与正常对照相比,SSc患者皮肤组织中CNN1的表达水平显著升高(P<0.05);皮肤组织中,CNN1的表达和α-SMA存在正相关,具有统计学意义(r=0.7219,P<0.0001);与正常对照组相比,SSc患者皮肤组织中CNN1的蛋白表达升高;原代皮肤成纤维细胞中,CNN1的表达和α-SMA、COL1A1均存在正相关,具有统计学意义(r=0.6547,P<0.05;r=0.6438,P<0.05);同时,原代皮肤成纤维细胞中干扰CNN1后,可显著抑制细胞增殖,降低纤维化相关基因(CTGF、COL1A2等)表达,降低胶原蛋白的表达。结论 CNN1在SSc患者中表达升高,且干扰CNN1可降低成纤维细胞活性,提示CNN1与SSc纤维化密切相关。
Objective To investigate the expression of calponin-1(CNN1) in systemic sclerosis(SSc) and its pathogenic role in fibrosis. Methods Skin biopsy samples were collected from 19 patients with SSc and 21 healthy subjects. Real-time PCR was used to detect the expression of CNN1 and α-SMA mRNAs in the samples, and the protein expression of CNN1 was detected using immunohistochemistry. In cultured primary human dermal fibroblasts, CNN1 expression was knocked down via RNA interference, and the mRNA expression levels of CNN1 and the fibrosis-related genes α-SMA, CTGF, COL1 A1, COL1 A2, and COL3 A1 were detected using real-time PCR;the proliferation of the cells was assessed using a real-time cell proliferation detection system. Results Compared with that in samples from normal subjects, the expression of CNN1 mRNA was significantly increased in the skin tissue of patients with SSc(P<0.05) with a positive correlation with α-SMA(r=0.7219, P<0.0001);the protein expression of CNN1 was also significantly increased in the skin tissue of patients with SSc. In cultured primary skin fibroblasts, the expression of CNN1 mRNA was positively correlated with α-SMA and COL1 A1 mRNA expressions(r=0.6547, P<0.05;r=0.6438, P<0.05). CNN1 knockdown in the fibroblasts significantly inhibited the cell proliferation, obviously lowered the expressions of fibrosis-related genes, and reduced the protein expression of collagen.Conclusion The expression of CNN1 is increased in the skin tissues of patients with SSc, and CNN1 knockdown can reduce the activity of dermal fibroblasts, suggesting the close correlation of CNN1 with fibrosis in SSc.
作者
赵晗
杨凯
刘庆梅
胡京晗
吴文育
王久存
ZHAO Han;YANG Kai;LIU Qingmei;HU Jinghan;WU Wenyu;WANG Jiucun(School of Life Sciences,Fudan University,Shanghai 200433,China;Department of Dermatology,Jing'an District Central Hospital,Shanghai 200040,China;Department of Dermatology,Huashan Hospital Affiliated to Fudan University,Shanghai 200040,China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2019年第3期279-285,共7页
Journal of Southern Medical University
基金
国家自然科学基金(81770066
81470254)
国家自然科学基金青年科学基金(81703117)~~
