胸腺素β4對人增生性瘢痕成纖維細胞膠原合成和CTGF表達的影響
發(fā)布時間:2018-08-15 17:13
【摘要】:目的: 擬通過觀察人增生性瘢痕成纖維細胞(hypertrophicscar fibroblasts HSFB)經(jīng)胸腺素β4(thymosin beta4)干預(yù)后,檢測其Ⅰ型和Ⅲ型膠原蛋白的生成水平及表達結(jié)締組織生長因子(connective tissue growth factor CTGF)的變化,以期了解胸腺素β4在病理性瘢痕發(fā)病中的作用機制。 方法: 標本來自本院整形科的患者(傷后半年內(nèi)),采用組織塊消化法培養(yǎng)細胞。成纖維細胞培養(yǎng)至3-6代細胞后進行實驗,共分成5組,其中空白對照組,加入無血清的培養(yǎng)基;實驗組:加入不同濃度的胸腺素β4(0.05、0.1、1、5μg/mL)無血清培養(yǎng)基作用培養(yǎng)6小時。(1)采用ELISA法檢測細胞上清液中Ⅰ型和Ⅲ型膠原蛋白的含量;(2)通過反轉(zhuǎn)錄-聚合酶鏈反應(yīng)(RT-PCR)法觀察結(jié)締組織生長因子(CTGF)mRNA的表達;(3)采用蛋白印跡法(Western blot)觀測CTGF蛋白的表達。 結(jié)果: (1)各實驗濃度的胸腺素β4作用6小時后Ⅰ型和Ⅲ型膠原蛋白分泌的值為:5μg/ml組(0.111±0.004、0.112±0.010),1μg/ml組(0.113±0.005、0.140±0.030),0.1μg/ml組(0.114±0.006、0.178±0.010),0.05μg/ml組(0.117±0.004、0.190±0.016),空白組(0.154±0.105,0.222±0.003),各實驗組與對照組差異有統(tǒng)計學(xué)意義(P0.05)。結(jié)果顯示,各實驗濃度胸腺素β4作用后增生性瘢痕成纖維細胞Ⅰ型和Ⅲ型膠原蛋白的分泌量減少。 (2)各實驗濃度的胸腺素β4作用6小時后(CTGF)mRNA表達灰度比值為:5μg/ml組(0.556±0.050),1μg/ml組(0.638±0.038),0.1μg/ml組(0.730±0.026),0.05μg/ml組(0.873±0.035),空白組(1.132±0.074),各實驗組與對照組差異有統(tǒng)計學(xué)意義(P0.05)。結(jié)果顯示,各實驗濃度(0.05、0.1、1、5μg/mL)胸腺素β4作用后增生性瘢痕成纖維細胞CTGFmRNA表達降低。 (3)各實驗濃度的胸腺素β4作用6小時后(CTGF)蛋白表達灰度比值為:5μg/ml組(0.510±0.044),,1μg/ml組(0.532±0.026),0.1μg/ml組(0.658±0.032),0.05μg/ml組(0.743±0.035),空白組(0.953±0.040),各實驗組與對照組差異有統(tǒng)計學(xué)意義(P0.05)。結(jié)果顯示,各實驗濃度(0.05、0.1、1、5μg/mL)胸腺素β4作用后均可以引起增生性瘢痕成纖維細胞CTGF蛋白表達降低。 結(jié)論: 胸腺素β4能劑量依賴性地降低增生性瘢痕成纖維細胞Ⅰ、Ⅲ型膠原蛋白的分泌及結(jié)締組織生長因子(CTGF)的表達。我們推認:胸腺素β4可能是通過這條途徑來影響瘢痕生成。
[Abstract]:Objective: to investigate the expression of connective tissue growth factor (connective tissue growth factor CTGF) in human hypertrophic scar fibroblasts (hypertrophicscar fibroblasts HSFB) after the intervention of thymosin 尾 4 (thymosin beta4). To understand the mechanism of thymosin 尾 4 in the pathogenesis of pathological scar. Methods: the specimens were collected from patients from plastic surgery department of our hospital (within six months after injury) and the cells were cultured by tissue mass digestion. The fibroblasts were cultured to 3-6 passages and were divided into five groups: blank control group and serum-free medium. Experimental group: thymosin 尾 _ 4 (0.05 ~ 0.1U ~ (-1) 渭 g/mL) was added to the serum free medium for 6 hours. (1) the content of type 鈪
本文編號:2184874
[Abstract]:Objective: to investigate the expression of connective tissue growth factor (connective tissue growth factor CTGF) in human hypertrophic scar fibroblasts (hypertrophicscar fibroblasts HSFB) after the intervention of thymosin 尾 4 (thymosin beta4). To understand the mechanism of thymosin 尾 4 in the pathogenesis of pathological scar. Methods: the specimens were collected from patients from plastic surgery department of our hospital (within six months after injury) and the cells were cultured by tissue mass digestion. The fibroblasts were cultured to 3-6 passages and were divided into five groups: blank control group and serum-free medium. Experimental group: thymosin 尾 _ 4 (0.05 ~ 0.1U ~ (-1) 渭 g/mL) was added to the serum free medium for 6 hours. (1) the content of type 鈪
本文編號:2184874
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