The effects of leptin downregulation by RNA interference on the proliferation ability transforming growth factor-β 1 and type Ⅰ collagen expression levels of pathological scar fibroblasts
10.3760/cma.j.cn114453-20200615-00367
- VernacularTitle:干扰小RNA下调瘦素的表达对人病理性瘢痕成纤维细胞增殖能力及转化生长因子-β 1、Ⅰ型胶原蛋白表达水平的影响
- Author:
Xiaojuan CHEN
1
;
Guofu ZHOU
;
Yan CAI
;
Xiaoyan XIANG
Author Information
1. 川北医学院附属医院烧伤整形外科，南充 637000
- Keywords:
Leptin;
Transforming growth factor-β;
TypeⅠ collagen;
Pathological scar;
Fibroblast;
Small interfering RNA
- From:
Chinese Journal of Plastic Surgery
2022;38(2):215-221
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of RNA interference on the expression level of leptin and its effect on proliferation, TGF-β 1 and collagen type Ⅰ production in human pathological scar fibroblasts in vitro. Methods:Pathological scar tissues (hypertrophic scar, keloid) were collected from the Department of Burn Plastic Surgery in Affiliated Hospital of North Sichuan Medical College. After primary cell culture and cell passage, passage 3 cells were selected for experimental study. The cells were divided into two groups: the experimental group which was transfected with leptin siRNA, and the negative control group which was transfected with empty vector. Examinations were carried out 48 hours after transfection. Cell proliferation was determined by CCK-8. The transcription levels of leptin, TGF-β 1 and type Ⅰ collagen genes were detected by polymerase chain reaction (PCR). The protein expression levels of leptin, TGF-β 1 and type Ⅰ collagen were determined by immunofluorescence and Western blotting. Student′s t-test was used for comparison between groups, and P<0.05 was considered statistically significant. Results:Five hypertrophic scars and five keloids were included. Compared with the negative control groups, the proliferation ability ( A450) of leptin-SiRNA transfected fibroblasts were not significantly different ( P>0.05). The relative mRNA expression levels of leptin, TGF-β 1 and type Ⅰ collagen in hypertrophic scars and keloids were significantly decreased in the siRNA transfection groups compared with the negative control groups ( P<0.05). Immunofluorescence results showed that the expression levels of leptin, TGF-β 1 and type I collagen in keloids were higher than those in hypertrophic scars, and that siRNA induced leptin downregulation significantly reduced the levels of leptin, TGF-β 1 and type I collagen in both hypertrophic scars and keloids. Western blotting showed that the protein levels of leptin, TGF-β 1 and type Ⅰ collagen were significantly decreased in hypertrophic scar and keloid fibroblasts after leptin siRNA interference ( P<0.05). Conclusions:Downregulation of leptin gene expression by RNA interference inhibited TGF-β 1 and typeⅠ collagen expression, which could be used in treating pathological scar.
