發(fā)布時(shí)間：2018-06-01 12:42

  本文選題：tmTNF-α + 急性鼠肝衰　； 參考：《華中科技大學(xué)》2014年博士論文

【摘要】：跨膜型TNF-α (Transmembrane tumor necrosis factor, tmTNF-α)是分泌型TNF-α (Secreted TNF-α, sTNF-α)前體,在TNF-α轉(zhuǎn)換酶處理后,釋放其胞外端而成為sTNF-α。兩型TNF-α均具有生物學(xué)功能。sTNF-α是典型的促炎細(xì)胞因子,在急性肝衰和內(nèi)毒素性休克發(fā)病機(jī)制中扮演著關(guān)鍵作用；然而,tmTNF-α在這兩種疾病中的作用雖然有轉(zhuǎn)基因動物的實(shí)驗(yàn)報(bào)道,但轉(zhuǎn)染的是缺失酶切位點(diǎn)的tmTNF-α突變體。我們的前期工作證實(shí)該突變體與野生型分子在信號轉(zhuǎn)導(dǎo)上有明顯差異,且不同實(shí)驗(yàn)室報(bào)道結(jié)果相反。鑒于這些報(bào)道可能不能反映天然tmTNF-α在這2種疾病中的作用,本研究旨在觀察內(nèi)源性tmTNF-α對急性鼠肝衰及內(nèi)毒素性休克的影響,并探索其作用機(jī)制。主要結(jié)果如下： 第一部分跨模型TNF-α對急性肝衰的影響 一、肝組織損傷變化特點(diǎn) 給小鼠注射LPS/D-gal制備急性肝衰模型,HE染色結(jié)果顯示,注射后2h可見肝細(xì)胞呈輕度水腫,4h肝細(xì)胞水腫,并有少量灶狀壞死,6h可見肝竇淤血,肝細(xì)胞壞死,肝細(xì)胞索斷裂；血清ALT和AST直至LPS/D-gal處理后6h才顯著升高,提示這個(gè)時(shí)間點(diǎn)肝組織明顯受損。 二、急性肝衰內(nèi)源性sTNF-α與tmTNF-α表達(dá)的變化與肝組織損傷的相關(guān)性 1. sTNF-α變化特點(diǎn)：ELISA結(jié)果顯示LPS/D-gal注射后2h小鼠血清sTNF-α達(dá)峰值,隨著時(shí)間延長而降低,4h后幾乎消失；從LPS/D-gal處理2h小鼠分離的KCs培養(yǎng)上清sTNF-α含量最高,而4h與6h分離的KCs培養(yǎng)上清sTNF-α雖然明顯降低,但仍然顯著高于正常鼠分離的KCs培養(yǎng)上清,提示循環(huán)中sTNF-α不能代表炎癥局部sTNF-α的變化。 2. tmTNF-α表達(dá)變化：用FCM分析急性肝衰小鼠KCs和PMs細(xì)胞表面tmTNF-α表達(dá)的變化,結(jié)果顯示在LPS/D-gal處理后2h及4h tmTNF-α呈低表達(dá),而在6htmTNF-α表達(dá)顯著上升。 3.兩型TNF-α與血清轉(zhuǎn)氨酶關(guān)系：急性肝衰小鼠血清TNF-α變化與血清轉(zhuǎn)氨酶AST及ALT變化相反,而KCs和PMs表達(dá)tmTNF-α水平與肝組織是否放至血清ALT或AST的量呈正相關(guān),提示是tmTNF-α而不是sTNF-α與急性肝組織損傷相關(guān)。 三、 KCs-6h與PMs-6h分泌促炎和抑炎因子失衡 1.血清促炎因子IL-6與抑炎因子IL-10的比值變化：用ELISA證實(shí)急性肝衰小鼠血清IL-6顯著升高,其在4h達(dá)高峰,在6h則有所降低,為高峰水平的87.8%；而血清IL-10含量則在2h最高,隨后下降,其在6h的水平僅為高峰水平的23.6%。計(jì)算IL-6與IL-10比值,發(fā)現(xiàn)其在6h才顯著增加,與肝組織損傷變化一致,提示血清IL-6/IL-10比值增加可更好的預(yù)測急性肝損傷。 2.高表達(dá)tmTNF-α的KCs-6h和PMs-6h分泌促炎和抑炎因子失衡：將LPS/D-gal注射后不同時(shí)間分離的KCs和PMs在體外進(jìn)行培養(yǎng),結(jié)果顯示這兩種巨噬細(xì)胞培養(yǎng)上清IL-6與IL-10的變化與血清變化一致；IL-6/IL-10比值在急性肝衰6h顯著增加,與此時(shí)間點(diǎn)這2種巨噬細(xì)胞高表達(dá)tmTNF-α呈正相關(guān)；提示在LPS/D-gal處理6h分離的KCs (KCs-6h)和PMs (PMs-6h)分泌促炎和抑炎因子嚴(yán)重失衡,進(jìn)而加重肝臟的炎性損傷。 四、高表達(dá)tmTNF-α的KCs-6h誘導(dǎo)肝細(xì)胞凋亡 1-KCs-6h通過直接接觸誘導(dǎo)肝細(xì)胞凋亡：將肝細(xì)胞與來自LPS/D-gal處理2h或6h的KCs進(jìn)行共培養(yǎng),結(jié)果顯示,高表達(dá)tmTNF-α的KCs-6h細(xì)胞可通過直接接觸誘導(dǎo)肝細(xì)胞凋亡增加,導(dǎo)致培養(yǎng)上清AST升高；而低表達(dá)tmTNF-α的KCs-2h細(xì)胞則無此效應(yīng)；當(dāng)用Transwell膜將KCs-6h細(xì)胞與肝細(xì)胞隔離培養(yǎng)后,無肝細(xì)胞出現(xiàn)凋亡。 2. KCs-6h細(xì)胞通過tmTNF-α直接誘導(dǎo)肝細(xì)胞凋亡：當(dāng)用TNF-α抗體預(yù)處理KCs-6h細(xì)胞30min,以中和其表達(dá)的tmTNF-α,再與肝細(xì)胞共培養(yǎng),肝細(xì)胞凋亡顯著降低,提示KCs-6h細(xì)胞表達(dá)的tmTNF-α可直接誘導(dǎo)肝細(xì)胞凋亡。 3.高表達(dá)tmTNF-α的KCs-6h細(xì)胞表達(dá)FasL增加：盡管用抗體中和tmTNF-α可顯著降低肝細(xì)胞凋亡,但仍有部分凋亡細(xì)胞,提示其它凋亡途徑可能參與了KCs-6h誘導(dǎo)肝細(xì)胞凋亡。Real time PCR和Western blotting的結(jié)果顯示高表達(dá)tmTNF-α的KCs-6h細(xì)胞表達(dá)FasL基因及其蛋白明顯高于低表達(dá)tmTNF-α的KCs-2h細(xì)胞,提示tmTNF-α可能通過增加FasL表達(dá)而促進(jìn)肝細(xì)胞凋亡。 五、過繼KCs-6h細(xì)胞可加重LPS/D-gal誘導(dǎo)小鼠肝組織損傷 1. KCs-6h細(xì)胞可持續(xù)表達(dá)tmTNF-α:為了保證過繼有效性,我們首先檢查KCs-6h細(xì)胞tmTNF-α表達(dá)是否穩(wěn)定。結(jié)果顯示：連續(xù)培養(yǎng)KCs-6h細(xì)胞至6天,其表達(dá)tmTNF-α仍能維持原有水平； 2.過繼KCs-6h細(xì)胞加重肝組織損傷：HE結(jié)果顯示：與LPS/D-gal組相比,過繼KCs-6h細(xì)胞可明顯加重小鼠在LPS/D-gal處理4h的肝組織損傷,表現(xiàn)為肝竇淤血,肝細(xì)胞索斷裂,水樣變性及少量壞死。 3.過繼KCs-6h促進(jìn)肝細(xì)胞凋亡：肝組織切片TUNEL染色顯示,過繼KCs-6h細(xì)胞明顯促進(jìn)肝細(xì)胞凋亡,血清AST也顯著高于過繼KCs-2h的小鼠,提示高表達(dá)tmTNF-α的KCs-6h在體內(nèi)可以加重LPS/D-gal誘導(dǎo)的肝衰。 4.過繼KCs-6h增加血清IL-6/IL-10比值：ELISA結(jié)果顯示過繼KCs-6h細(xì)胞導(dǎo)致血清促炎因子IL-6和IL-1β含量明顯高于過繼KCs-2h組小鼠,而抑炎因子IL-10低于KCs-2h組小鼠,IL-6/IL-10比值明顯增加,提示高表達(dá)tmTNF-α的KCs-6h分泌促炎和抑炎因子平衡障礙,從而參與急性肝衰的發(fā)生發(fā)展。 第二部分跨模型TNF-α對內(nèi)毒素性休克的影響 一、特異性tmTNF Ab的制備與鑒定： 1.tmTNFAb的制備與鑒定：針對tmTNF-α單表位成功制備了兔抗小鼠tmTNF-α抗體。ELISA顯示兔血清含有高效價(jià)抗tmTNF-α單表位抗體,且只結(jié)合tmTNF-α肽段,不與sTNF-α發(fā)生交叉反應(yīng)。 2. tmTNF Ab可與細(xì)胞表面tmTNF-α結(jié)合：用FCM證實(shí)該抗體可與TNF-α+NH3T3細(xì)胞表面tmTNF-α結(jié)合,而不能與TNF-α陰性的CT26與H22細(xì)胞株結(jié)合,提示該抗體可識別細(xì)胞表面完整的tmTNF-α分子。 二、tmTNF Ab可有效阻斷tmTNF-α轉(zhuǎn)換為sTNF-α: 1. tmTNF Ab阻斷tmTNF-α轉(zhuǎn)換為sTNF-α:用100ng/ml LPS和tmTNF Ab分別刺激RAW264.7巨噬細(xì)胞系和PMs原代細(xì)胞4h, ELISA和FCM結(jié)果顯示該抗體可有效抑制sTNF-α分泌,而顯著增加tmTNF-α的表達(dá),提示該抗體可有效阻斷tmTNF-α轉(zhuǎn)換為sTNF-α。 2. tmTNF Ab抑制LPS誘導(dǎo)RAW264.7上清sTNF-α胞毒效應(yīng)：用生物活性證實(shí)LPS刺激RAW264.74h的上清對TNF-α敏感靶細(xì)胞L929具有明顯胞毒效應(yīng),用TNF-α抗體預(yù)先中和上清sTNF-α后,可阻斷其胞毒效應(yīng)；用tmTNF Ab明顯抑制LPS刺激RAW264.7上清的胞毒效應(yīng),再次證實(shí)tmTNF Ab抑制1tmTNF-α酶解,致使sTNF-α分泌減少,因而其胞毒效應(yīng)受到抑制。 四、tmTNF Ab對LPS誘導(dǎo)巨噬細(xì)胞產(chǎn)生促炎和抑炎因子的影響 1. tmTNF Ab抑制LPS誘導(dǎo)巨噬細(xì)胞產(chǎn)生NO：用LPS和tmTNF Ab刺激RAW264.7和PMs,結(jié)果顯示該抗體可明顯抑制LPS誘導(dǎo)這2種巨噬細(xì)胞轉(zhuǎn)錄iNOS基因以及分泌NO。 2. tmTNF Ab抑制LPS誘導(dǎo)巨噬細(xì)胞產(chǎn)生促炎因子IL-6和IL-1β：用LPS和tmTNF Ab刺激RAW264.7和PMs,結(jié)果顯示該抗體可明顯抑制LPS誘導(dǎo)這2種巨噬細(xì)胞轉(zhuǎn)錄IL-6和IL-1p基因及分泌這些細(xì)胞因子。 3. tmTNF Ab對LPS誘導(dǎo)巨噬細(xì)胞產(chǎn)生抑炎因子IL-10無影響：用LPS和tmTNFAb刺激RAW264.7和PMs,結(jié)果顯示該抗體對LPS誘導(dǎo)這2種巨噬細(xì)胞轉(zhuǎn)錄IL-10基因及其表達(dá)無影響。 五、tmTNF Ab保護(hù)小鼠抵抗內(nèi)毒素血癥 1. tmTNF Ab可阻斷內(nèi)毒素血癥小鼠tmTNF-α轉(zhuǎn)變?yōu)閟TNF-α:預(yù)先給予tmTNF-α Ab明顯增強(qiáng)LPS注射小鼠4h腹腔巨噬細(xì)胞的tmTNF-α表達(dá),同時(shí)抑制該時(shí)間點(diǎn)內(nèi)毒素血癥小鼠血清TNF-α含量(P0.01),提示tmTNF Ab在體內(nèi)可有效阻斷tmTNF-α轉(zhuǎn)換成sTNF-α。 2. tmTNF Ab可改善內(nèi)毒素性休克臨床癥狀,提高小鼠存活率：用tmTNF Ab可明顯減輕內(nèi)毒素性休克鼠唇紫紺,改善臨床評分,顯著提高內(nèi)毒素血癥小鼠的存活率。 3. tmTNF Ab抑制內(nèi)毒素血癥促炎因子分泌：LPS注射小鼠2h血清促炎因子IL-6、IL-1p及NO以及抑炎因子IL-10即開始升高,6h達(dá)高峰,此后下降。用tmTNF Ab可明顯降低內(nèi)毒素血癥小鼠上述血清促炎因子的水平,卻輕度增加IL-10含量,但無統(tǒng)計(jì)學(xué)意義。 綜上所述,tmTNF-α在不同的病理狀態(tài)下發(fā)揮不同的、甚至相反的作用。在LPS/D-gal誘導(dǎo)的急性鼠肝衰中KCs和PMs表達(dá)的tmTNF-α可直接誘導(dǎo)肝細(xì)胞凋亡以及通過分泌促炎/抗炎因子失衡加重肝臟的炎性損傷。反之,用tmTNF Ab促進(jìn)tmTNF-α表達(dá),抑制sTNF-α分泌,在體外可抑制巨噬細(xì)胞對LPS的應(yīng)答,在體內(nèi)保護(hù)小鼠抵抗內(nèi)毒素性休克。該研究不但證實(shí)tmTNF-α在不同疾病中的作用,還為治療TNF相關(guān)疾病提供新的策略和線索。
[Abstract]:Transmembrane type TNF- alpha (Transmembrane tumor necrosis factor, tmTNF- a) is the precursor of secretory TNF- alpha (Secreted TNF- alpha, sTNF- alpha). After TNF- alpha converting enzyme treatment, it releases its extracellular end and becomes sTNF- alpha. Type two alpha has biological function, which is a typical proinflammatory cytokine, in acute liver failure and endotoxic shock pathogenesis. However, although the role of tmTNF- alpha in these two diseases has been reported in transgenic animals, the transfection is a tmTNF- alpha mutant with the deletion of the enzyme cut site. Our previous work proved that the mutant and the wild type molecules have obvious differences in signal transduction, and the results of different laboratory reports are opposite. Given that these reports may not reflect the role of natural tmTNF- alpha in these 2 diseases, this study aims to observe the effect of endogenous tmTNF- a on acute hepatic failure and endotoxic shock in rats and explore its mechanism. The main results are as follows:
Part one the effect of trans model TNF- alpha on acute liver failure
The characteristics of the changes of liver tissue injury
The mice were injected with LPS/D-gal to prepare the acute liver failure model. The results of HE staining showed that the liver cells showed mild edema, 4H hepatocyte edema, and a small amount of focal necrosis, 6h showed hepatic sinusoid congestion, hepatocyte necrosis, and hepatic cell cord fracture, and the serum ALT and AST until LPS/ D-gal treatment was significantly elevated, suggesting this time point liver group. The fabric was obviously damaged.
Two, the correlation between the expression of endogenous sTNF- alpha and tmTNF- alpha in acute liver failure and liver tissue injury.
1. sTNF- alpha change characteristics: ELISA results showed that sTNF- alpha reached peak value in 2H mice after LPS/D-gal injection, decreased with time and almost disappeared after 4H. STNF- alpha content was the highest in KCs culture supernatant from LPS/D-gal treated 2H mice, while 4H and 6h isolated culture supernatant decreased significantly, but still significantly higher than normal mice. The isolated KCs culture supernatant indicated that sTNF- alpha in the circulation could not represent the change of local sTNF- alpha in inflammation.
2. tmTNF- alpha expression changes: the changes in the expression of tmTNF- alpha on the surface of KCs and PMs cells in acute liver failure mice were analyzed with FCM. The results showed that the expression of 2H and 4h tmTNF- alpha was low after LPS/D-gal treatment, and the expression of 6htmTNF- a was significantly increased.
The relationship between 3. type two TNF- alpha and serum transaminase: the changes of serum TNF- a in mice with acute liver failure are contrary to the changes of serum aminotransferase AST and ALT, while the level of KCs and PMs expression tmTNF- A is positively correlated with the amount of ALT or AST in the liver tissue, suggesting that tmTNF- a rather than sTNF- alpha is associated with acute liver tissue injury.
Three, KCs-6h and PMs-6h secrete an imbalance between pro inflammatory and anti inflammatory factors.
1. the change of the ratio of serum proinflammatory factor IL-6 and anti inflammatory factor IL-10: the serum IL-6 in mice with acute liver failure was significantly increased with ELISA, which was at the peak of 4h, and decreased in 6h to the peak level, while the serum IL-10 content was highest in 2H, then decreased, and the ratio of 23.6%. IL-6 to IL-10 in 6h was only at the peak level of 23.6%.. It was found that it increased significantly in 6h, which was consistent with the change of liver tissue injury, suggesting that the increase of serum IL-6/IL-10 ratio can better predict acute liver injury.
2. high expression of KCs-6h and PMs-6h secretion of tmTNF- A and the imbalance of proinflammatory and anti inflammatory factors: KCs and PMs, isolated at different time after LPS/D-gal injection, were cultured in vitro. The results showed that the changes of IL-6 and IL-10 in the supernatant of the two macrophages were in accordance with the serum changes; the IL-6/IL-10 ratio in acute liver failure 6h increased significantly, and this time point was found. These 2 macrophages have a positive correlation with high expression of tmTNF- - alpha, suggesting that KCs (KCs-6h) and PMs (PMs-6h) secreted by 6h and PMs (PMs-6h) secretes a serious imbalance of proinflammatory and anti inflammatory factors in LPS/D-gal, and thus aggravates the inflammatory injury of the liver.
Four, high expression of tmTNF- alpha KCs-6h induces hepatocyte apoptosis.
1-KCs-6h induced hepatocyte apoptosis through direct contact: co culture of hepatocytes with KCs from LPS/D-gal to treat 2H or 6h. The results showed that KCs-6h cells with high expression of tmTNF- alpha could induce apoptosis in liver cells by direct contact, leading to the increase of AST in culture supernatant, while KCs-2h cells with low level of tmTNF- alpha did not have this effect; T After ranswell membrane isolated KCs-6h cells from hepatocytes, no hepatocytes apoptosis.
2. KCs-6h cells directly induced the apoptosis of hepatocytes through tmTNF- alpha: when the KCs-6h cell 30min was pretreated with TNF- alpha antibody, the expression of tmTNF- a was neutralized, and the hepatocytes were co cultured with the hepatocytes, and the apoptosis of the liver cells decreased significantly. It suggested that the tmTNF- a expressed by KCs-6h cells could directly induce the apoptosis of the liver cells.
3. the expression of FasL in KCs-6h cells with high expression of tmTNF- alpha was increased: Although there was a significant decrease in apoptosis of liver cells with antibodies and tmTNF- a, there were still some apoptotic cells, suggesting that other apoptotic pathways may be involved in KCs-6h induced apoptosis of hepatocytes,.Real time PCR and Western blotting, showing the expression of KCs-6h cells expressing high expression tmTNF- alpha. The gene and its protein were significantly higher than those of KCs-2h cells with low expression of tmTNF- alpha, suggesting that tmTNF- alpha might promote hepatocyte apoptosis by increasing FasL expression.
Five, adoptive KCs-6h cells can aggravate LPS/D-gal induced liver injury in mice.
1. KCs-6h cells express tmTNF- alpha sustainably: in order to ensure the adoptive effectiveness, we first examine the stability of the expression of tmTNF- alpha in KCs-6h cells. The results show that the expression of tmTNF- a still can maintain the original level of the KCs-6h cells in continuous culture to 6 days.
2. adoptive KCs-6h cells aggravated liver tissue damage: HE results showed that adoptive KCs-6h cells significantly aggravated the liver tissue damage in LPS/D-gal treatment of 4H in mice, manifested as hepatic sinusoid congestion, hepatic cell cord rupture, water like degeneration and a small amount of necrosis.
3. adoptive KCs-6h promoted hepatocyte apoptosis: liver tissue section TUNEL staining showed that adoptive KCs-6h cells obviously promoted hepatocyte apoptosis, and serum AST was significantly higher than that of adoptive KCs-2h mice, suggesting that KCs-6h with high expression of tmTNF- alpha could aggravate LPS/D-gal induced liver failure in the body.
4. adoptive KCs-6h increased the serum IL-6/IL-10 ratio: ELISA results showed that the serum proinflammatory factor IL-6 and IL-1 beta in the adoptive KCs-6h cell was significantly higher than that of the adoptive KCs-2h group, while the anti inflammatory factor IL-10 was lower than the KCs-2h group, and the IL-6/IL-10 ratio was significantly increased, suggesting that the KCs-6h secreting proinflammatory and anti inflammatory factor balance barrier of high expression tmTNF- alpha was high. It is involved in the development of acute liver failure.
The second part is the effect of trans model TNF- alpha on endotoxic shock.
1. Preparation and identification of specific tmTNF Ab:
Preparation and identification of 1.tmTNFAb: a Rabbit anti mouse tmTNF- alpha antibody.ELISA was successfully prepared for the single epitope of tmTNF- a to show that the rabbit serum contains high effective anti tmTNF- alpha epitopes antibody and only tmTNF- alpha peptide segment, and no cross reaction with sTNF- alpha.
2. tmTNF Ab can bind to tmTNF- alpha on cell surface: it is confirmed by FCM that the antibody can bind to tmTNF- alpha on the surface of TNF- alpha +NH3T3 cells, but can not be combined with TNF- alpha negative CT26 and H22 cell lines, suggesting that the antibody can identify the intact tmTNF- alpha molecule on the surface of the cell.
Two, tmTNF Ab can effectively block tmTNF- to sTNF- alpha.
1. tmTNF Ab blocked tmTNF- alpha to sTNF- alpha: 100ng/ml LPS and tmTNF Ab stimulated the RAW264.7 macrophage system and PMs primary cell 4H. ELISA and results showed that the antibody could effectively inhibit the secretion of alpha, which significantly increased the expression of alpha alpha, suggesting that the antibody could effectively obstruct the conversion of alpha to alpha.
2. tmTNF Ab inhibited the cytotoxic effect of sTNF- alpha induced by LPS in RAW264.7 supernatant: using bioactivity to confirm that LPS stimulated RAW264.74h in the supernatant of RAW264.74h had obvious cytotoxic effect on TNF- alpha sensitive target cell L929, and the cytotoxic effect could be blocked by using TNF- alpha antibody in advance to neutralize sTNF- alpha, and the cytotoxic effect of stimulating supernatant was inhibited by tmTNF. It is confirmed that tmTNF Ab inhibits 1tmTNF- alpha enzymolysis, resulting in a decrease in sTNF- alpha secretion, thus inhibiting its cytotoxic effect.
Four, the effect of tmTNF Ab on LPS induced macrophages producing proinflammatory and anti-inflammatory factors.
1. tmTNF Ab inhibited LPS induced macrophage to produce NO: LPS and tmTNF Ab were used to stimulate RAW264.7 and PMs. The results showed that the antibody inhibited LPS to induce the iNOS gene of the 2 macrophages and secreted NO..
2. tmTNF Ab inhibited LPS induced macrophages to produce proinflammatory cytokines IL-6 and IL-1 beta: LPS and tmTNF Ab stimulated RAW264.7 and PMs. The results showed that the antibody inhibited LPS induction of the 2 macrophages transcriptional IL-6 and genes and secreting these cytokines.
3. tmTNF Ab had no effect on LPS induced macrophage induced anti inflammatory factor IL-10: RAW264.7 and PMs were stimulated with LPS and tmTNFAb. The results showed that the antibody had no effect on LPS induced 2 macrophage transcription IL-10 gene and its expression.
Five, tmTNF Ab protects against endotoxemia in mice
1. tmTNF Ab could block the transformation of tmTNF- alpha into sTNF- alpha in mice with endotoxemia: pre administration of tmTNF- alpha Ab significantly enhanced the tmTNF- alpha expression of 4H peritoneal macrophages in LPS injected mice, and inhibited the content of TNF- alpha in serum of mice with endotoxemia at the same time (P0.01).
2. tmTNF Ab can improve the clinical symptoms of endotoxic shock and improve the survival rate of mice: using tmTNF Ab can significantly reduce cyanosis in endotoxic shock rats, improve the clinical score, and significantly improve the survival rate of endotoxemia mice.
3. tmTNF Ab inhibited the secretion of proinflammatory cytokines from endotoxemia: LPS injection of 2H serum proinflammatory factor IL-6, IL-1p and NO and anti inflammatory factor IL-10 began to rise, and 6h reached the peak and decreased thereafter. TmTNF Ab could significantly reduce the levels of serum proinflammatory factors in mice with endotoxemia, but slightly increased IL-10 content, but no statistical significance.
To sum up, tmTNF- alpha plays different, even opposite effects in different pathological conditions. In the acute rat liver failure induced by LPS/D-gal, the tmTNF- alpha expressed by KCs and PMs can directly induce hepatocyte apoptosis and aggravate the inflammatory damage of the liver by secreting proinflammatory / anti-inflammatory factor imbalance. On the other hand, tmTNF Ab promotes the expression of tmTNF- A and inhibits sT. NF- alpha secretion, which inhibits the response of macrophages to LPS in vitro, protects mice against endotoxic shock in the body. This study not only confirms the role of tmTNF- alpha in different diseases, but also provides new strategies and clues for the treatment of TNF related diseases.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2014
【分類號】：R459.7;R575.3

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 于孟學(xué);曹金;李薇;;生物制劑治療系統(tǒng)性紅斑狼瘡的新進(jìn)展[J];北京醫(yī)學(xué);2008年01期

2 黃艷;黃剛;胡長江;曾益軍;許志臻;陳\宏，

本文編號：1964221