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移植脂肪来源干细胞对兔耳增生性瘢痕组织中成纤维细胞α-SMA和DCN表达的影响。

Effects of transplanted adipose derived stem cells on the expressions of α-SMA and DCN in fibroblasts of hypertrophic scar tissues in rabbit ears.

作者信息

Chu Haihan, Wang Yunpeng, Wang Xiuchun, Song Xianhui, Liu Huaqing, Li Xue

机构信息

Department of Burn and Plastic Surgery, Jining First People's Hospital, Jining, Shandong 272000, P.R. China.

Department of Dermatology, Rizhao Hospital of Dermatology, Rizhao, Shandong 276800, P.R. China.

出版信息

Exp Ther Med. 2018 Sep;16(3):1729-1734. doi: 10.3892/etm.2018.6383. Epub 2018 Jun 29.

Abstract

To study the effects of transplanted adipose derived stem cells (ADSCs) on the expressions of α-smooth muscle actin (α-SMA) and decorin (DCN) in fibroblasts of hypertrophic scar tissues in rabbit ears. Twelve New Zealand white rabbits were selected; the normal subcutaneous adipose tissues in inguinal region were removed, ADSCs were extracted via enzyme digestion, cultured in Dulbecco's modified Eagle's medium (DMEM) and inoculated into the culture dish (3-5×10 cells/ml). After the rabbit ear hypertrophic scar model was established successfully, the fibroblasts of hypertrophic scar tissues in rabbit ears were separated and cultured using the mechanical method combined with enzyme digestion, and the ADSCs and scar fibroblasts were cultured in non-contact Transwell co-culture system for 21 days (experimental group); the corresponding scar fibroblasts were cultured in an ordinary 6-well plate without any treatment for 21 days (control group). The content of collagen I in fibroblasts was detected using the enzyme-linked immunosorbent assay (ELISA) kit, the mRNA expressions of α-SMA and DCN were detected via reverse transcription-polymerase chain reaction (RT-PCR), the protein expressions of α-SMA and DCN were detected via western blot analysis, and the expressions and distribution of α-SMA and DCN were detected via immunofluorescence assay. The results of ELISA showed that the content of collagen I in experimental group was decreased significantly (p<0.01). The results of RT-PCR and western blot analysis revealed that the mRNA and protein expression levels of α-SMA were significantly decreased (P<0.01, but those of DCN were significantly increased (p<0.01). Moreover, the results of immunofluorescence assay showed that the expression of α-SMA in experimental group was significantly decreased, while the expression of DCN was significantly increased. ADSCs can inhibit the mRNA and protein expressions of α-SMA and promote the mRNA and protein expressions of DCN in culture system, and they are expected to be used in the prevention and treatment of pathological scars.

摘要

研究移植脂肪来源干细胞(ADSCs)对兔耳增生性瘢痕组织成纤维细胞中α-平滑肌肌动蛋白(α-SMA)和核心蛋白聚糖(DCN)表达的影响。选取12只新西兰白兔;去除腹股沟区正常皮下脂肪组织,通过酶消化法提取ADSCs,在杜氏改良 Eagle培养基(DMEM)中培养并接种于培养皿(3 - 5×10个细胞/毫升)。成功建立兔耳增生性瘢痕模型后,采用机械法联合酶消化法分离并培养兔耳增生性瘢痕组织的成纤维细胞,将ADSCs与瘢痕成纤维细胞在非接触式Transwell共培养系统中培养21天(实验组);相应的瘢痕成纤维细胞在普通6孔板中不做任何处理培养21天(对照组)。使用酶联免疫吸附测定(ELISA)试剂盒检测成纤维细胞中I型胶原的含量,通过逆转录-聚合酶链反应(RT-PCR)检测α-SMA和DCN的mRNA表达,通过蛋白质印迹分析检测α-SMA和DCN的蛋白表达,通过免疫荧光测定检测α-SMA和DCN的表达及分布。ELISA结果显示,实验组I型胶原含量显著降低(p<0.01)。RT-PCR和蛋白质印迹分析结果显示,α-SMA的mRNA和蛋白表达水平显著降低(P<0.01),而DCN的则显著升高(p<0.01)。此外,免疫荧光测定结果显示,实验组α-SMA的表达显著降低,而DCN的表达显著升高。ADSCs在培养体系中可抑制α-SMA的mRNA和蛋白表达,促进DCN的mRNA和蛋白表达,有望用于病理性瘢痕的防治。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6a12/6122172/9499e8f33efa/etm-16-03-1729-g00.jpg

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