本文選題：原代枯否細胞 + 干擾素調(diào)節(jié)因子��； 參考：《中國免疫學(xué)雜志》2016年04期

【摘要】：目的:探討干擾素調(diào)節(jié)因子3(Interferon regulator factor 3,IRF3)shRNA腺病毒對脂多糖(Lipopolysaccharide,LPS)刺激枯否細胞(Kupffer cell,KC)早期細胞因子分泌動態(tài)變化的影響。方法:采用在體灌注分離培養(yǎng)大鼠原代KC,以IRF3shRNA腺病毒體外感染KC,48 h后采用LPS刺激細胞,于0、2、4和6 h收集細胞培養(yǎng)上清液,并收集6 h細胞。上清液細胞因子的分泌采用ELISA分析;細胞IRF3基因表達采用RT-PCR和Western blot方法檢測。結(jié)果:LPS刺激誘導(dǎo)了KC內(nèi)IRF3 mRNA和蛋白質(zhì)表達升高,IRF3 shRNA腺病毒的應(yīng)用抑制了LPS刺激誘導(dǎo)和非刺激組成性IRF3 mRNA和蛋白質(zhì)的表達;KC受LPS刺激活化后極早期(2 h)IFN-β分泌即上升,4 h達峰值,6 h分泌水平開始下降,但仍維持于高水平。干擾腺病毒的應(yīng)用抑制了LPS刺激后各時間點IFN-β的分泌,抑制分泌高峰的出現(xiàn),并使6 h分泌水平趨于正常;KC活化后極早期即分泌大量TNF-α,并于2 h內(nèi)達到峰值,隨后分泌逐漸下降,但6 h仍維持于高水平。干擾腺病毒的應(yīng)用抑制了LPS刺激各時間點TNF-α的分泌,并抑制了分泌高峰的出現(xiàn);IL-1β分泌增高出現(xiàn)于LPS刺激4 h后,6 h分泌水平達更高值。干擾腺病毒的應(yīng)用抑制了LPS刺激早期KC對IL-1β的分泌;KC受LPS刺激活化后極早期IL-10分泌即上升,且隨著LPS刺激時間延長,其分泌水平逐漸增加。IRF3 shRNA腺病毒的應(yīng)用促進了LPS刺激后早期各時間點IL-10的分泌。結(jié)論:IRF3 shRNA腺病毒可使原代枯否細胞IRF3基因表達沉默;在LPS刺激原代KC,IRF3可促進其下游信號分子IFN-β、前炎細胞因子TNF-α和IL-1β的分泌,并抑制抑炎細胞因子IL-10分泌。因此,IRF3可能在肝組織免疫炎癥性損傷的發(fā)生中起中心作用。
[Abstract]:Aim: to investigate the effect of interferon regulatory factor 3(Interferon regulator factor 3 (IRF3) shRNA adenovirus on the dynamic changes of cytokine secretion in Kupffer cell (KC) stimulated by lipopolysaccharide (LPS). Methods: primary rat KC cells were isolated and cultured by perfusion in vivo. The cells were stimulated by LPS for 48 h after infection with IRF3shRNA adenovirus in vitro. The supernatants were collected at 4 and 6 hours after infection with IRF3shRNA adenovirus, and the cells were collected for 6 h. The secretion of cytokines in supernatant was analyzed by ELISA and the expression of IRF3 gene was detected by RT-PCR and Western blot. Results the expression of IRF3 mRNA and protein in KC was induced by 1: LPS-stimulated. The application of IRF3 shRNA adenovirus inhibited the expression of IRF3 mRNA and protein in KC induced by LPS and non-stimulative components. The secretion of IFN- 尾 increased at the very early stage after the activation of KC stimulated by LPS. The level of secretion began to decrease at 4 h and 6 h. But it remains at a high level. The application of interfering adenovirus inhibited the secretion of IFN- 尾 at different time points after stimulation of LPS, inhibited the peak of secretion, and made the secretion level of 6 h tend to secrete a large amount of TNF- 偽 in the early stage after normal activation of KC, and reached the peak value within 2 h. Then the secretion decreased gradually, but remained at a high level at 6 h. The application of interfering adenovirus inhibited the secretion of TNF- 偽 at different time points stimulated by LPS, and inhibited the peak of secretion. The increase of IL-1 尾 secretion occurred at 6 h after LPS stimulation. The application of interfering adenovirus inhibited the secretion of IL-1 尾 by KC in the early stage of LPS stimulation. After activation by LPS, the secretion of IL-10 increased in the very early stage, and with the prolongation of the time of LPS stimulation, the secretion of KC increased in the early stage. Its secretion level increased gradually. IRF3 shRNA adenovirus promoted the secretion of IL-10 at different time points after LPS stimulation. Conclusion the expression of IRF3 gene in Kupffer cells was silenced by the adenovirus of 1: IRF3 shRNA, which stimulated the secretion of IFN- 尾, proinflammatory cytokines TNF- 偽 and IL-1 尾, and inhibited the secretion of anti-inflammatory cytokine IL-10 in primary Kupffer cells by LPS stimulation. Therefore, IRF3 may play a central role in the development of immune inflammatory injury in liver tissue.
【作者單位】： 南京醫(yī)科大學(xué)附屬上海松江中心醫(yī)院/上海交通大學(xué)附屬第一人民醫(yī)院松江分院感染科;
【基金】：國家自然科學(xué)基金項目(81070357,30660066) 上海市松江區(qū)科學(xué)技術(shù)攻關(guān)項目(14SJGGYY22)
【分類號】：R575.3

【相似文獻】

相關(guān)期刊論文 前10條

1 孫慶;鄧存良;;枯否細胞的生物學(xué)功能與肝細胞損傷[J];西南軍醫(yī);2008年02期

2 劉敏芝;;慢性肝病時枯否細胞的消減與肝癌發(fā)生的關(guān)系[J];國外醫(yī)學(xué)(內(nèi)科學(xué)分冊);1984年06期

3 張海燕;劉立新;韓德五;;急性壞死性胰腺炎時枯否細胞吞噬功能的實驗研究[J];山西醫(yī)科大學(xué)學(xué)報;2007年05期

4 張海燕;劉立新;韓德五;;急性壞死性胰腺炎大鼠枯否細胞分泌功能的實驗研究[J];山西醫(yī)科大學(xué)學(xué)報;2007年11期

5 李海;肝臟內(nèi)皮細胞和枯否細胞的細胞生物學(xué)[J];國外醫(yī)學(xué)(消化系疾病分冊);1995年04期

6 張海燕;劉立新;韓德五;;枯否細胞抑制致急性壞死性胰腺炎大鼠微循環(huán)改變的實驗研究[J];山西醫(yī)科大學(xué)學(xué)報;2007年09期

7 孫文兵;韓本立;李昆;彭志明;段恒春;馬瑞亮;陳娟;王槐志;;老化肝臟枯否細胞骨架結(jié)構(gòu)及其對內(nèi)毒素劑量耐受性的改變[J];中華肝臟病雜志;1997年01期

8 吳惠文;許瑞齡;;枯否細胞在高脂飲食致小鼠慢性肝損傷發(fā)生中的作用[J];胃腸病學(xué)和肝病學(xué)雜志;2007年06期

9 向德棟,李奇芬,王宇明,雷虹;維生素E對大鼠肝臟枯否細胞NF-κB的影響[J];重慶醫(yī)學(xué);2002年02期

10 柳俊剛;高楓;張森;楊劍鋒;劉傳淵;;BALB/c鼠肝臟枯否細胞不同分離培養(yǎng)方法的比較[J];結(jié)直腸肛門外科;2009年05期

相關(guān)會議論文 前2條

1 鞏義春;李聞;Leung;陳志偉;鐘尚志;;流式細胞分離技術(shù)純化肝臟枯否細胞新方法[A];中華醫(yī)學(xué)會第七次全國消化病學(xué)術(shù)會議論文匯編（下冊）[C];2007年

2 孟瑩;李聞;楊云生;;膽汁內(nèi)外引流術(shù)對梗阻性黃疸大鼠血清TNF-α和肝臟枯否細胞誘導(dǎo)型一氧化氮合酶表達的影響[A];中華醫(yī)學(xué)會第七次全國消化病學(xué)術(shù)會議論文匯編（下冊）[C];2007年

相關(guān)博士學(xué)位論文 前3條

1 ，

本文編號：1943239