Study on construction of recombinant plasmid carrying the Decorin gene and its effects on hypertrophic scars of rabbit ears
10.3760/cma.j.cn114453-20190522-00165
- VernacularTitle:携带Decorin基因的质粒DNA构建及其对兔耳增生性瘢痕的治疗作用
- Author:
Jiali YANG
1
;
Yanxiang WANG
;
Li DU
;
Yuxin LU
;
Jiabing WU
;
Xiaochen CHENG
;
Ziqiang PAN
;
Xirong ZHAO
;
Qinglin ZHANG
Author Information
1. 淮阴工学院生命科学与食品工程学院，淮安 223003；军事科学院军事医学研究院辐射医学研究所，北京 100850
- Keywords:
Decorin;
Recombinant plasmid;
Hypertrophic scar;
Transforming growth factor-β1
- From:
Chinese Journal of Plastic Surgery
2020;36(10):1121-1127
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct a recombinant plasmid DNA carrying the decorin( DCN) gene and study its therapeutic effect on hypertrophic scars of rabbit ears. Methods:The human decorin gene fragment amplified by PCR was cloned into plasmid vector pUDK to construct the recombinant plasmid pDCN, which was identified by enzyme digestion and sequencing. pDCN was transfected into 293T cells, and the expression of DCN and TGF-β1 was detected. The therapeutic effect of pDCN on rabbit ear hypertrophic scar model was observed by hypertrophy index, pathological examination and immunohistochemical staining. Results:The decorin gene was successfully inserted into pUDK, which was examined by enzyme digestion and sequencing. The expression level of mRNA and protein of DCN was up-regulated in 293T cells post pDCN transfection, and the expression of TGF-β1 was suppressed. Then the rabbit ear hypertrophic scars were treated with different doses of pDCN, and the results showed that the hypertrophy index of the medium dose (200 μg/cm 2) pDCN group was significantly lower than that of the PBS group ( P<0.05), while there was no significant difference in the hypertrophy index of the low dose and high dose pDCN group compared with the PBS group. The expression of DCN in ears skin in the medium dose pDCN group was significantly higher than that in the PBS group ( P<0.05). The pathological examination showed that inflammatory cell infiltration and fibrous deposition in scar tissue were significantly reduced. These results indicated that the medium-dose pDCN could effectively inhibit the hyperplasia of hypertrophic scar in rabbit ears. Conclusions:pDCN, the plasmid carrying decorin gene, has therapeutic effects on hypertrophic scars of rabbit ears by inhibiting TGF-β1 expression.
