[1]王润芝,吴皓,刘一鸣,等.阻断Kupffer细胞IRE1-XBP1通路对大鼠肝移植排斥反应的影响[J].第三军医大学学报,2016,38(22):2431-2437.
 Wang Runzhi,Wu Hao,Liu Yiming,et al.Effects of inhibiting IRE1-XBP1 pathway in Kupffer cells in rats after liver transplant rejection [J].J Third Mil Med Univ,2016,38(22):2431-2437.
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阻断Kupffer细胞IRE1-XBP1通路对大鼠肝移植排斥反应的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
38卷
期数:
2016年第22期
页码:
2431-2437
栏目:
基础医学
出版日期:
2016-11-30

文章信息/Info

Title:
Effects of inhibiting IRE1-XBP1 pathway in Kupffer cells in rats after liver transplant rejection
 
作者:
王润芝吴皓刘一鸣曹丁吴涯昆李金政龚建平
重庆医科大学附属第二医院肝胆外科
Author(s):
Wang Runzhi Wu Hao Liu Yiming Cao Ding Wu Yakun Li Jinzheng Gong Jianping

Department of Hepatobiliary Surgery, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, China

关键词:
肌醇酶1X盒蛋白连接蛋白1Kupffer细胞肝移植急性排斥反应
Keywords:
inositol enzyme 1 X-box binding protein 1 Kupffer cells liver transplantation acute rejection
分类号:
R392.4;R617;R657.3
文献标志码:
A
摘要:

目的      探讨阻断Kupffer细胞IREXBP1通路对大鼠原位肝移植排斥反应的作用。方法     术前用GdCl3处理或未处理建立大鼠原位肝移植急性排斥反应模型,未处理大鼠随机数字法分为XBP1-shRNA组(A组),Scrambled-shRNA组(B组),PBS组(C组);处理大鼠随机分为GdCl3+XBP1-shRNA组(D组),GdCl3+ScrambledshRNA组(E组),GdCl3+PBS组(F组)。未处理组术后检测血清ALT、AST、IFNγ、IL-10、IL-17的表达水平;RT-PCR检测T-bet、RORγt、IFN-γ、IL-17及IL-10 mRNA水平;Western blot检测CD86、CD206、IFNγ、IL-17及IL-10蛋白表达水平,HE染色观察肝组织结构,根据Banff方案进行RAI评分,各组剩余大鼠观察术后生存率。处理组术后检测IFN-γ、IL-17及IL-10 mRNA以及蛋白表达水平,HE染色观察肝组织结构。结果     在未处理组中,与B、C组比较,A组各时间点肝功能指标ALT、AST明显下降(P<0.05),A组血清表达IFN-γ、IL-10明显受到抑制(P<0.05),IL-10的表达水平则呈明显上升趋势(P<0.05),与RT-PCR和Western blot结果趋势相符合,另外A组CD86蛋白表明显下降(P<0.05),而表达CD206上升(P<0.05),Tbet/RORγt mRNA相对表达量也明显下降(P<0.05),B、C组上述指标与A组对比则呈相反趋势,而且B、C组之间比较差异无统计学意义(P>0.05);A组RAI评分(3.83±0.14),明显低于B、C组RAI评分(9.13±0.20)、(8.95±026)(P<0.05),并且A组大鼠的生存时间明显高于B、C组大鼠(P<0.05)。在处理组中,D、E和F组肝脏局部IFN-γ、IL-17、IL-10 mRNA和蛋白表达情况以及病理组织AcR程度比较无统计学上的差异(P>0.05)。结论     阻断IRE1-XBP1通路促进了KCs的M1型极化,引起Th1/Th17向Th2/Treg的免疫偏移,在一定程度上减轻了移植肝脏急性排斥反应的程度。

Abstract:

Objective      To investigate the effects of inhibiting inositol requiring 1-X-Box binding protein 1 (IRE1-XBP1) pathway in Kupffer cells (KCs) on liver transplant rejection in rats.   Methods       Orthotopic liver transplantation in rats was established with or without GdCl3 treatment pre-operation. In untreated group, the rats were divided randomly into 3 subgroups, that is, XBP1-shRNA group (A), Scrambled-shRNA group (B), and PBS group (C). Likewise, the treated group was sub-divided into GdCl3+XBP1-shRNA group (D), GdCl3+Scrambled-shRNA group (E), and GdCl3+PBS group (F). In untreated group,  the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), interferon-γ (IFN-γ), interleukin-10 (IL-10) and IL-17 were examined at different time points after operation. Liver tissues were excised for hematoxylin and eosin (HE) staining, mRNA expression levels of transcriptional factors (T-bet and RORγt) and cytokines (IFN-γ, IL-17 and IL-10) quantified by real-time PCR (RT-PCR), and protein levels of CD86, CD206, IFN-γ, IL-17 and IL-10 by Western blotting. Banff schema was used for grading the acute allograft rejection with rejection activity index (RAI). The survival rate was assessed by Kaplan-Meier curve analysis. In the treated group, the mRNA and the protein expression of IFN-γ, IL-17 and IL-10 was also tested. Results     In the untreated group, the serum levels of ALT and AST at different time points were significantly lower in group A compared with groups B and C (P<0.05). In group A, IFN-γ and IL-17 were significantly decreased in serum while IL-10 was significantly increased (P<0.05). There were consistent with the results of RT-PCR and western blotting. In addition, the relative mRNA expression of T-bet/RORγt was decreased, protein expression of CD86 decreased and CD206 increased, which were all statistically significant (P<0.05). In contrast, groups B and C showed inverse trend with respect to the above-mentioned factors, mRNA and protein expressions. Moreover, the differences between group B and C were not significant. The mean RAI score in group A was 3.81±0.14, which was significantly lower than that in groups B (9.13±0.20) and C (8.95±0.26), respectively (P<0.05). The data was also in agreement with the Kaplan-Meier curve, showing that rats in group A significantly survived longer than these in groups B and C (P<0.05). However, the expression of IFN-γ, IL-17 and IL-10 both at mRNA and protein levels was not significantly different among the treatment groups D, E, and F. The same results in pathological examination were observed.  Conclusion       The blockage of IRE1-XBP1 pathway could alter the polarization of KCs from M1 to M2 type, lead to skewing of Th1/Th17 towards Th2/Treg, and consequently ameliorate acute rejection following liver transplantation in rats.

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更新日期/Last Update: 2016-11-23