本文選題：組織工程　切入點(diǎn)：肝細(xì)胞　出處：《中國組織工程研究》2015年51期

【摘要】：背景:肝星狀細(xì)胞是肝內(nèi)分泌細(xì)胞外基質(zhì)的關(guān)鍵細(xì)胞類型,因此在肝纖維化研究中很重要。目的:實(shí)驗(yàn)改良了肝星狀細(xì)胞原代培養(yǎng)方法,獲得充足的細(xì)胞量,用轉(zhuǎn)化生長因子β1刺激活化,研究致纖維化的相關(guān)因子表達(dá)情況。方法:氯胺酮麻醉小鼠后采用門靜脈穿刺原位灌注肝臟的方法分離肝臟,采用密度梯度離心獲得肝星狀細(xì)胞,利用形態(tài)學(xué)、免疫熒光染色等方法對(duì)原代肝星狀細(xì)胞進(jìn)行鑒定;將培養(yǎng)24 h肝星狀細(xì)胞用質(zhì)量濃度10μg/L轉(zhuǎn)化生長因子β1刺激48 h,同時(shí)設(shè)PBS組進(jìn)行對(duì)比。結(jié)果與結(jié)論:通過原位灌注肝臟并且酶消化的方法可以成功獲得小鼠原代肝星狀細(xì)胞,并且錐蟲藍(lán)染色活率為(97.2±0.8)%,免疫熒光染色純度為(90.4±1.2)%,細(xì)胞計(jì)數(shù)總量約2.5×106/只。實(shí)時(shí)熒光定量聚合酶鏈?zhǔn)椒磻?yīng)檢測(cè)顯示,與PBS組比較,轉(zhuǎn)化生長因子β1刺激組的肝星狀細(xì)胞平滑肌肌動(dòng)蛋白α、Ⅰ型膠原蛋白、轉(zhuǎn)化生長因子β受體1、轉(zhuǎn)化生長因子β受體2 mR NA表達(dá)水平明顯升高(P0.05)。結(jié)果證實(shí),實(shí)驗(yàn)采用的改良的培養(yǎng)方法可更高效高質(zhì)量的獲得原代肝星狀細(xì)胞,轉(zhuǎn)化生長因子β1刺激可導(dǎo)致肝星狀細(xì)胞活化,分泌致纖維化因子。
[Abstract]:Background: hepatic stellate cells (HSCs) are the key cell types of extracellular matrix (ECM) of liver, so they are very important in the study of hepatic fibrosis.Aim: to improve the primary culture method of hepatic stellate cells (HSC), obtain sufficient cells, stimulate activation with transforming growth factor 尾 1 (TGF- 尾 1), and study the expression of fibrosis related factors.Methods: after ketamine anesthetized mice liver was isolated by portal vein puncture in situ perfusion. Hepatic stellate cells were obtained by density gradient centrifugation. Primary hepatic stellate cells were identified by means of morphology and immunofluorescence staining.Hepatic stellate cells were cultured for 24 h and stimulated with 10 渭 g / L TGF- 尾 1 for 48 h, and compared with PBS group.Results and conclusion: primary hepatic stellate cells of mice could be successfully obtained by in situ perfusion of liver and enzyme digestion. The viability of trypanosome blue staining was 97.2 鹵0.8. The purity of immunofluorescence staining was 90.4 鹵1.2. The total number of cells was about 2.5 脳 106 per mouse.Real-time fluorescence quantitative polymerase chain reaction showed that the hepatic stellate cell smooth muscle actin 偽, type I collagen were stimulated by transforming growth factor 尾 1 (TGF- 尾 1) compared with PBS group.The expression of transforming growth factor 尾 receptor 1 and transforming growth factor 尾 receptor 2mR na increased significantly (P 0.05).The results showed that the improved culture method could obtain the primary hepatic stellate cells more efficiently and qualitatively, and the stimulation of transforming growth factor 尾 1 could lead to the activation of hepatic stellate cells and the secretion of fibrogenic factors.
【作者單位】： 新疆醫(yī)科大學(xué)第一附屬醫(yī)院/臨床醫(yī)學(xué)研究院 新疆重大疾病醫(yī)學(xué)重點(diǎn)實(shí)驗(yàn)室-省部共建國家重點(diǎn)實(shí)驗(yàn)室培育基地 新疆包蟲病基礎(chǔ)醫(yī)學(xué)重點(diǎn)實(shí)驗(yàn)室;新疆醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院生物化學(xué)與分子生物學(xué)教研室;
【基金】：國家自然科學(xué)基金面上項(xiàng)目(81371838) 新疆維吾爾自治區(qū)自然科學(xué)基金青年項(xiàng)目(2015211C084)~~
【分類號(hào)】：R575.2

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