發(fā)布時(shí)間：2018-05-09 13:39

  本文選題：增生性瘢痕 + 轉(zhuǎn)化生長因子β1　； 參考：《暨南大學(xué)》2015年碩士論文

【摘要】：研究背景增生性瘢痕是深度燒傷常見的并發(fā)癥,病理表現(xiàn)為大量的成纖維細(xì)胞增殖及細(xì)胞外基質(zhì)(ECM)的異常沉積。轉(zhuǎn)化生長因子β1(Transforming growth factorβ1,TGF-β1)是一種具有多種功能的細(xì)胞因子,通過細(xì)胞內(nèi)信號轉(zhuǎn)導(dǎo)發(fā)揮作用,在增生性瘢痕的形成過程中扮演重要角色:它一方面能調(diào)節(jié)成纖維細(xì)胞生長、分化、遷移凋亡;另一方面促進(jìn)成纖維細(xì)胞合成、分泌膠原蛋白,抑制基質(zhì)金屬蛋白酶(MMPs)合成,減少膠原纖維降解。芍藥苷(Paeoniflorin)是傳統(tǒng)中藥芍藥(Paeonia lactiflora pall)干燥根中的提取物,也是芍藥的主要單體活性成分。研究發(fā)現(xiàn),芍藥苷在肝、腎、肺中表現(xiàn)出抗纖維化的作用,其主要機(jī)制為減少組織內(nèi)TGF-β1的含量,抑制細(xì)胞內(nèi)TGF-β1的自分泌,使膠原等細(xì)胞外基質(zhì)的合成明顯減少。增生性瘢痕是皮膚創(chuàng)傷后發(fā)生在真皮的纖維增生性疾病,而芍藥苷對增生性瘢痕的形成的影響,尚未見報(bào)道。目的探討芍藥苷對增生性瘢痕成纖維細(xì)胞增殖及其生物學(xué)行為的影響。方法收集增生性瘢痕組織及來自同一個(gè)體的正常皮膚組織,采用組織塊法分離培養(yǎng)增生性瘢痕成纖維細(xì)胞和正常皮膚成纖維細(xì)胞,分別加入不同濃度(0,200,400,800μM)的芍藥苷:(1)培養(yǎng)24h、48h、72h,MTS法檢測細(xì)胞增殖速度;(2)培養(yǎng)48h,流式細(xì)胞儀檢測細(xì)胞周期及凋亡;(3)培養(yǎng)48h,收集細(xì)胞培養(yǎng)上清液,酶聯(lián)免疫吸附分析方法(ELISA)檢測TGF-β1、I型膠原含量;(4)培養(yǎng)48h,蛋白質(zhì)印跡法(Western Blot)檢測細(xì)胞中α平滑肌肌動(dòng)蛋白(α-SMA)的表達(dá)水平。結(jié)果(1)濃度為400μM、800μM的芍藥苷能明顯抑制各時(shí)相點(diǎn)(24h、48h、72h)增生性瘢痕及正常皮膚成纖維細(xì)胞的增殖。(2)200μM、400μM、800μM的芍藥苷能明顯增加兩種成纖維細(xì)胞處于G2期的百分比,各組中未檢測到細(xì)胞凋亡。(3)400μM、800μM的芍藥苷能明顯減少增生性瘢痕成纖維細(xì)胞培養(yǎng)上清液中TGF-β1含量,但不影響正常皮膚成纖維細(xì)胞培養(yǎng)上清液中TGF-β1含量。(4)200μM、400μM、800μM的芍藥苷能明顯減少增生性瘢痕及正常皮膚成纖維細(xì)胞上清液中I型膠原含量。(5)200μM、400μM、800μM的芍藥苷能明顯減少增生性瘢痕及正常皮膚成纖維細(xì)胞中α-SMA蛋白的表達(dá)。結(jié)論一定濃度的芍藥苷可抑制增生性瘢痕及正常皮膚成纖維細(xì)胞的增殖,減少I型膠原的分泌及α-SMA的表達(dá);芍藥苷可以減少增生性瘢痕成纖維細(xì)胞TGF-β1的自分泌,但是對正常皮膚成纖維細(xì)胞TGF-β1的自分泌卻沒有明顯的影響,提示芍藥苷可能對于增生性瘢痕的形成有抑制作用。
[Abstract]:Background Hypertrophic scar is a common complication of deep burn. The pathological features of hypertrophic scar are proliferation of fibroblasts and abnormal deposition of extracellular matrix (ECM). Transforming growth factor 尾 1(Transforming growth factor 尾 1 (TGF- 尾 1) is a multifunctional cytokine that plays an important role in the process of hypertrophic scar formation through intracellular signal transduction: on the one hand, it can regulate the growth and differentiation of fibroblasts. Migration and apoptosis; on the other hand, promote fibroblast synthesis, secrete collagen, inhibit matrix metalloproteinase (MMPs) synthesis, reduce collagen degradation. Paeoniflorin (Paeoniflorin) is an extract from the dried roots of Paeonia lactiflora pallin, a traditional Chinese medicine, which is also the main monomer active component of paeoniflorin. It was found that paeoniflorin had anti-fibrosis effect in liver, kidney and lung. The main mechanism of paeoniflorin was to reduce the content of TGF- 尾 _ 1 in tissue, inhibit the autocrine of TGF- 尾 _ 1 and decrease the synthesis of extracellular matrix such as collagen. Hypertrophic scar is a fibrous hyperplastic disease occurring in the dermis after skin trauma, but the effect of paeoniflorin on the formation of hypertrophic scar has not been reported. Objective to investigate the effects of paeoniflorin on proliferation and biological behavior of hypertrophic scar fibroblasts. Methods the hypertrophic scar tissue and the normal skin tissue from the same body were collected. The fibroblasts of hypertrophic scar and normal skin were isolated and cultured by tissue block method. Paeoniflorin 1 of different concentrations (0 200 400 800 渭 M) was added to culture for 24 h, 48 h and 72 h, respectively. The cell proliferation rate was detected by MTS method. The cell cycle and apoptosis were detected by flow cytometry (FCM) for 48 h. The supernatants of cell culture were collected. The level of 偽 -SMAs was detected by Elisa and Western blotts were used to detect the expression of 偽 -SMAin in TGF- 尾 _ 1C _ I type I collagen (TGF- 尾 _ (1) I) cultured for 48 h. The expression of 偽 -SMAs was detected by Western blot assay (Western blot). The expression of 偽 -SMAs was detected by Western blot assay. Results Paeoniflorin at the concentration of 400 渭 M and 800 渭 M could significantly inhibit the proliferation of hypertrophic scar and normal skin fibroblasts at 24 h and 48 h / 72 h) Paeoniflorin at the concentration of 400 渭 M or 800 渭 M could significantly increase the percentage of the two fibroblasts in G2 phase. No apoptosis was detected in each group. Paeoniflorin (800 渭 M) could significantly reduce the content of TGF- 尾 1 in the culture supernatant of hypertrophic scar fibroblasts. However, Paeoniflorin of 200 渭 M, 400 渭 M, 800 渭 M of paeoniflorin in the supernatant of normal skin fibroblast culture could significantly reduce the content of collagen type I in the supernatant of hypertrophic scar and fibroblast supernatant of normal skin, and the content of paeoniflorin in the supernatant of normal skin fibroblast cell culture. 200 渭 M, 400 渭 M, 800 渭 M of paeoniflorin, could significantly reduce the proliferative ability of paeoniflorin. Expression of 偽 -SMA protein in fibroblasts of scar and normal skin. Conclusion Paeoniflorin can inhibit the proliferation of hypertrophic scar and normal skin fibroblasts, decrease the secretion of type I collagen and the expression of 偽 -SMA, and decrease the autocrine of TGF- 尾 1 in hypertrophic scar fibroblasts. However, there was no obvious effect on the autosecretion of TGF- 尾 1 in normal skin fibroblasts, suggesting that paeoniflorin might inhibit the formation of hypertrophic scar.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R622

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 胡宗濤;高世樂;秦峰;朱杰;甄毅嵐;董六一;;芍藥苷對大鼠放射性肝纖維化的保護(hù)作用和機(jī)制研究[J];解放軍藥學(xué)學(xué)報(bào);2012年04期

2 孫麗榮;曹雄;侯鳳青;朱心紅;高天明;;芍藥苷研究進(jìn)展[J];中國中藥雜志;2008年18期

3 儲德勇;李叢磊;楊楓;吳強(qiáng);李靜;丁向東;羅慶禮;沈繼龍;;芍藥苷對日本血吸蟲感染小鼠肝組織免疫病理的影響[J];中國寄生蟲學(xué)與寄生蟲病雜志;2008年01期

，

本文編號：1866224