本文選題：黃芩苷 + 肺成纖維細(xì)胞��； 參考：《河北醫(yī)科大學(xué)》2013年碩士論文

【摘要】：目的：肺纖維化的病理特點是肺肌成纖維細(xì)胞異常增多，過度的細(xì)胞外基質(zhì)沉積。α-平滑肌肌動蛋白（smooth muscle actin, α-SMA)是肌成纖維細(xì)胞（myofibroblast，MFB）的標(biāo)志物。轉(zhuǎn)化生長因子-β1（transformin growthfactor-beta，TGF-β1）是刺激成纖維細(xì)胞向肌成纖維細(xì)胞轉(zhuǎn)化的主要細(xì)胞因子。結(jié)締組織生長因子（connective tissue growth factor,CTGF）是TGF-β1的下游因子。肺成纖維細(xì)胞轉(zhuǎn)化為肌成纖維細(xì)胞時，CTGF表達(dá)明顯增加。黃芩苷(Baicalin)是從中草藥黃芩中分離出來的一種黃酮類化合物，具有抗纖維化作用。但黃芩苷對于離體肺成纖維細(xì)胞和肌成纖維細(xì)胞CTGF及α-SMA蛋白表達(dá)的影響目前尚未見報道。因此，本實驗通過離體培養(yǎng)大鼠肺成纖維細(xì)胞及肌成纖維細(xì)胞，觀察黃芩苷對肺成纖維細(xì)胞及肺肌成纖維細(xì)胞CTGF及α-SMA蛋白表達(dá)的影響，，為黃芩苷在臨床肺纖維化中的防治提供研究資料。 方法:體外分離培養(yǎng)原代雄性SD大鼠(150-170g)肺成纖維細(xì)胞；用TGF-β誘導(dǎo)肺成纖維細(xì)胞轉(zhuǎn)化為肌成纖維細(xì)胞；MTT法測定黃芩苷對肺成纖維細(xì)胞及肌成纖維細(xì)胞存活率的影響；Western Blot法測定肺成纖維細(xì)胞及肌成纖維細(xì)胞CTGF及α-SMA的蛋白表達(dá)。 結(jié)果： 1黃芩苷對肺成纖維細(xì)胞及肌成纖維細(xì)胞存活率的影響 肺成纖維細(xì)胞是指正常培養(yǎng)的無TGF-β誘導(dǎo)的肺成纖維細(xì)胞，肌成纖維細(xì)胞為肺成纖維細(xì)胞經(jīng)TGF-β誘導(dǎo)48h且α-SMA表達(dá)增加的細(xì)胞，對照組為無藥物干擾的。與對照組(0μmol/L)相比，10、20和40μmol/L的黃芩苷對肺成纖維細(xì)胞及肌成纖維細(xì)胞的存活率均無明顯影響（P0.05）,80、160μmol/L濃度的黃芩苷抑制肺成纖維細(xì)胞及肌成纖維細(xì)胞的存活率（P0.01）。提示10-40μmol/L濃度的黃芩苷對肺成纖維細(xì)胞及肌成纖維細(xì)胞無明顯毒性作用。 2黃芩苷對肺成纖維細(xì)胞CTGF及α-SMA蛋白表達(dá)的影響 與對照組(0μmol/L)相比,10、20、40μmol/L濃度的黃芩苷對肺成纖維細(xì)胞CTGF及α-SMA蛋白表達(dá)均無統(tǒng)計學(xué)意義（P＞0.05），80、160μmol/L濃度的黃芩苷則抑制肺成纖維細(xì)胞CTGF及α-SMA蛋白表達(dá)（P＜0.01）。這提示10-40μmol/L黃芩苷既不增加肺成纖維細(xì)胞CTGF表達(dá)也不誘導(dǎo)其向肌成纖維細(xì)胞轉(zhuǎn)化，80-160μmol/L黃芩苷則抑制其表達(dá)。 3黃芩苷對肺肌成纖維細(xì)胞CTGF及α-SMA蛋白表達(dá)的影響 TGF-β誘導(dǎo)后的細(xì)胞與正常肺纖維細(xì)胞相比，CTGF及α-SMA蛋白表達(dá)明顯升高，提示體外肌成纖維細(xì)胞誘導(dǎo)成功。與對照組相比(0μmol/L)，10、20、40、80、160μmol/L濃度的黃芩苷呈劑量依賴性的抑制肌成纖維細(xì)胞CTGF及α-SMA蛋白表達(dá)（P＜0.01），提示黃芩苷呈劑量依賴性促進(jìn)肌成纖維細(xì)胞去分化為成纖維細(xì)胞。 結(jié)論： 110-40μmol/L濃度的黃芩苷對肺成纖維細(xì)胞及肌成纖維細(xì)胞存活率無影響，80、160μmol/L濃度的黃芩苷則表現(xiàn)出毒性作用。 310-40μmol/L濃度的黃芩苷既不增加肺成纖維細(xì)胞CTGF表達(dá)也不誘導(dǎo)其向肌成纖維細(xì)胞轉(zhuǎn)化，80-160μmol/L濃度的黃芩苷抑制肺成纖維細(xì)胞CTGF及α-SMA蛋白表達(dá)。 4黃芩苷呈劑量依賴性的（10、20、40、80、160μmol/L）抑制肌成纖維細(xì)胞CTGF表達(dá)，并促進(jìn)其去分化為成纖維細(xì)胞。
[Abstract]:Objective: the pathological characteristics of pulmonary fibrosis are abnormal increase of pulmonary myofibroblast and excessive extracellular matrix deposition. Alpha smooth muscle actin (smooth muscle actin, alpha -SMA) is a marker of myofibroblast (myofibroblast, MFB). Transforming growth factor - beta 1 (transformin growthfactor-beta, TGF- beta 1) is a stimulant to fibroblast cells. The main cytokine of the transformation of myofibroblast. Connective tissue growth factor (CTGF) is the downstream factor of TGF- beta 1. When pulmonary fibroblasts are converted into myofibroblast, the expression of CTGF is significantly increased. Baicalin (Baicalin) is a kind of flavonoids separated from the Chinese herbal medicine baicalin. However, the effect of Baicalin on the expression of CTGF and alpha -SMA protein in isolated lung fibroblasts and myofibroblasts has not yet been reported. Therefore, the experiment was conducted to observe the effects of Baicalin on pulmonary fibroblast and pulmonary myofibroblast CTGF and alpha -SMA protein by cultured rat lung fibroblasts and myofibroblast in vitro. The influence of Baicalin on prevention and treatment of clinical pulmonary fibrosis is provided.
Methods: the primary male SD rat (150-170g) lung fibroblasts were isolated and cultured in vitro, and the pulmonary fibroblasts were transformed into myofibroblast with TGF- beta, and the effects of Baicalin on the survival rate of pulmonary fibroblasts and myofibroblasts were measured by MTT, and the Western Blot method was used to determine the CTGF and alpha -SMA of pulmonary fibroblasts and myofibroblasts. Protein expression.
Result錛

本文編號：1920342