本文選題：kupffer細(xì)胞　切入點(diǎn)：內(nèi)毒素耐受　出處：《中山大學(xué)學(xué)報(bào)(醫(yī)學(xué)科學(xué)版)》2017年05期 　論文類型：期刊論文

【摘要】：【目的】探討上調(diào)pellino-1在Kupffer細(xì)胞(KC)內(nèi)毒素耐受時(shí)對(duì)腫瘤壞死因子受體相關(guān)因子3(TRAF3)泛素化、促分裂原活化蛋白激酶(MAPK)信號(hào)通路及下游細(xì)胞因子分泌情況的影響�！痉椒ā糠蛛x、培養(yǎng)C57BL/6小鼠KC,隨機(jī)分為2組:(1)空載對(duì)照組:轉(zhuǎn)染空載質(zhì)粒48 h后,先給予小劑量LPS(10 ng/mL)刺激24 h,再給予大劑量LPS(300 ng/mL)刺激。(2)過(guò)表達(dá)組:過(guò)表達(dá)pellino-1慢病毒轉(zhuǎn)染48 h后,先給予小劑量LPS(10 ng/mL)刺激24 h,再給予大劑量LPS(300 ng/mL)刺激。兩組分別于處理后0、5、10、30、60 min收獲細(xì)胞,蛋白質(zhì)免疫印跡試驗(yàn)(Western blot)檢測(cè)pellino-1、K48泛素化(K48ubiquitin,K48-Ub)、TRAF3、JNK、p-JNK、p38、p-p38蛋白水平表達(dá)變化;酶聯(lián)免疫吸附法(ELISA)檢測(cè)細(xì)胞培養(yǎng)上清液中IL-1β、TNF-α及IL-10的分泌情況。【結(jié)果】與空載對(duì)照組相比,過(guò)表達(dá)組pellino-1蛋白表達(dá)量在各個(gè)時(shí)間點(diǎn)均明顯升高;K48-Ub水平明顯升高;TRAF3蛋白表達(dá)量明顯降低;JNK、p38蛋白表達(dá)量沒(méi)有明顯變化,但p-JNK及p-p38蛋白表達(dá)量顯著升高;過(guò)表達(dá)組細(xì)胞上清中IL-1β及TNF-α的量明顯升高(P0.05),而IL-10的量則明顯降低(P0.05)�！窘Y(jié)論】過(guò)表達(dá)pellino-1可促進(jìn)TRAF3蛋白K-48泛素化降解,導(dǎo)致TRAF3蛋白表達(dá)量降低,激活下游的MAPK信號(hào),從而抑制內(nèi)毒素耐受的形成。
[Abstract]:[objective] to investigate the effect of upregulation of pellino-1 on Ubiquitin, mitogen-activated protein kinase (MAPK) signaling pathway and cytokine secretion in Kupffer cells during endotoxin tolerance. The cultured C57BL / 6 mice were randomly divided into two groups: control group: after 48 h of transfection of empty plasmid, the mice were stimulated with small dose of LPS(10 ng / mL for 24 h, and then stimulated with large dose of LPS(300 ng / mL. (2) overexpression of pellino-1: 48 h after transfection of pellino-1 lentivirus. Small dose of LPS(10 ng / mL was given for 24 h, and then high dose of LPS(300 ng / mL). The expression of pellino-1k48 ubiquitin K48-UbC3TRAF3KNKp-JNKP p38 p38 protein was detected by Western blotting in the two groups at 0 ~ 5 ~ 10 ~ 10 ~ 30 ~ 60 min ~ (-1) ~ (th) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (?). Enzyme linked immunosorbent assay (Elisa) was used to detect the secretion of IL-1 尾 -TNF- 偽 and IL-10 in the supernatant of cell culture. In the overexpression group, the expression of pellino-1 protein increased significantly at each time point, the level of K48-Ub increased significantly, the expression of TRAF3 protein decreased significantly, but the expression of p-JNK and p-p38 protein increased significantly. In overexpression group, the amount of IL-1 尾 and TNF- 偽 in supernatant increased significantly, while the amount of IL-10 decreased significantly. [conclusion] overexpression of pellino-1 can promote the degradation of K-48 ubiquitin of TRAF3 protein, decrease the expression of TRAF3 protein and activate the downstream MAPK signal. Thus inhibiting the formation of endotoxin tolerance.
【作者單位】： 重慶醫(yī)科大學(xué)附屬第二醫(yī)院肝膽外科;重慶醫(yī)科大學(xué)附屬第一醫(yī)院肝膽外科;
【基金】：國(guó)家自然科學(xué)青年基金(81401622;81301656) 重慶市科委基礎(chǔ)科學(xué)與前沿技術(shù)研究專項(xiàng)(重點(diǎn),cstc2015 jcyjBX0070)
【分類號(hào)】：R459.7
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本文編號(hào)：1579328