The Third Military Medical University Southwest Plastic Surgery Hospital, Shapingba District, Chongqing, China.
Eur Rev Med Pharmacol Sci. 2017 May;21(10):2512-2517.
To study the regulatory mechanism of miR-29 over TGF-β1 and COL1 in scar cells.
5 clinical cases of hypertrophic scar (HS) skin and adjacent normal skin tissues were separated into fibroblast for primary culture and subculture before being observed morphologically and standard HE staining under an ordinary optical microscope. RT-PCR method was applied to test the expression level of miR-29, TGF-β1, and COL1 mRNA. ELISA method was applied to test the expression level of extracellular matrix COL1, fibronectin (FN) and α-SMA. The miR-29 overexpression vector was built and transfected in vitro. RT-PCR method was applied to test related genes and ELISA method was applied to test the expression level of the extracellular matrix.
The color of karyon and cytoplasm of normal fibroblast were both light red, with little ECM. The color of karyon of scar fibroblast was blue. The cytoplasm was red of different degrees, with relatively much ECM, in deep blue color. Compared with that in the normal fibroblast group, the miR-29 mRNA in fibroblast in the scar group significantly decreased (p<0.05). The TGF-β1 and COL1 mRNA significantly increased (p<0.05). The COL1, FN and α-SMA level were significantly higher (p<0.05) than that in the normal group. These mRNAs levels in miR-29 overexpression group were lower than scar group but higher than the normal group.
The expression of miR-29 which regulates the expression of TGF-β1 and COL1 and increases the level of ECM significantly decreases in scar cells. This one suggests a mechanism of the formation of the scars through TGF-β1 and COL1.
研究 miR-29 对瘢痕成纤维细胞中 TGF-β1 和 COL1 的调控机制。
分离 5 例增生性瘢痕（HS）皮肤及其相邻正常皮肤组织，原代培养和传代培养成纤维细胞，普通光镜下观察形态，行标准 HE 染色。采用 RT-PCR 法检测 miR-29、TGF-β1、COL1 mRNA 的表达水平，采用 ELISA 法检测细胞外基质 COL1、纤维连接蛋白（FN）和α-SMA 的表达水平。构建 miR-29 过表达载体并进行体外转染，采用 RT-PCR 法检测相关基因，采用 ELISA 法检测细胞外基质的表达水平。
正常成纤维细胞的核和细胞质颜色均为浅红色，细胞外基质较少。瘢痕成纤维细胞核呈蓝色，细胞质呈不同程度的红色，呈深蓝色，细胞外基质较多。与正常成纤维细胞组比较，瘢痕成纤维细胞中 miR-29 mRNA 明显降低（p<0.05），TGF-β1 和 COL1 mRNA 明显升高（p<0.05），COL1、FN 和α-SMA 水平明显升高（p<0.05），高于正常组。miR-29 过表达组的这些 mRNAs 水平低于瘢痕组，但高于正常组。
miR-29 的表达下调，调控 TGF-β1 和 COL1 的表达，显著增加 ECM 水平，在瘢痕细胞中明显降低。这表明 TGF-β1 和 COL1 通过形成瘢痕的机制之一。
