【摘要】：目的：研究慢病毒介導(dǎo)RNA干擾小鼠供心al,3GT、NF-κB表達后異種移植物的存活時間與免疫病理變化。 方法：采用改良Heron技術(shù)套袖法建立小鼠到大鼠異種異位心臟移植模型(小鼠心臟移植到大鼠右側(cè)頸部)。按照RNA干擾供心靶基因不同分組：對照組、空病毒組、RNAiα1,3GT組、RNAi NF-κB組、RNAα1,3GT+NF-κB組,每組8對。觀察移植供心存活時間,并使用SABC法檢測失功心臟補體C3、IgM、IgG、NK細胞、巨噬細胞、ICAM-1表達情況。 結(jié)果：對照組、空病毒組、RNAiα1,3GT組、RNAiNF-κB組、RNAiα1,3GT+NF-κB組異種移植物的平均存活時間分別為(32±2.5)h,(31±2.5)h,(51±4.0)h,(31±1.8)h,(53±7.3)h.對照組、空病毒組、RNAiNF-κB組三組移植物生存時間無明顯差別(p0.05),RNAial,3GT組、RNAial,3GT+NF-κB組移植物生存時間較對照組、空病毒組、RNAiNF-κB組明顯延長(p0.05), RNAiα1,3GT組、RNAiα1,3GT+ NF-κB組兩組移植物生存時間無明顯差別(p0.05)。 SABC法檢測補體C3、IgM、IgG在對照組、空病毒組、RNAi NF-κB組三組均可見大量沉積,且三組間無明顯差別(p0.05)。RNAiα1,3GT組、RNAiα1,3GT+NF-κB組補體C3、IgM、IgG沉積較對照組、空病毒組、RNAiNF-KB組減少(p0.05), RNAiα1,3GT組、RNAiα1,3GT+NF-κB組兩組間補體C3、IgM、IgG沉積無明顯差別(p0.05)；NK細胞、巨噬細胞、ICAM-1在對照組、空病毒組、RNAiNF-KB組三組有陽性表達,且三組間無明顯差別(p0.05),在RNAiα1，3GT組、RNAiα1,3GT+NF-κB組浸潤表達較對照組、空病毒組、RNAiNF-κB組增加(p0.05),且RNAiα1,3GT+NF-κB組較RNAiα1,3GT組減少(p0.05)。 結(jié)論：異種抗原Galα(1,3)Gal在小鼠到大鼠異種心臟移植中起重要作用,RNAi可減少其表達使移植物存活時間延長,使異種移植物中補體C3、IgN、IgG沉積減少,巨噬細胞、自然殺傷細胞浸潤增加,細胞間黏附分子-1表達增加,呈現(xiàn)為由HAR向AVR轉(zhuǎn)變的趨勢。小鼠到大鼠異種移植排斥具有HAR為主且與AVR共存的特點。RNAi NF-κB,可使移植物巨噬細胞、自然殺傷細胞浸潤減少,細胞間黏附分子表達減低,可能減輕異種AVR反應(yīng)。
[Abstract]:Aim: to study the survival time and immunopathologic changes of xenografts after lentivirus-mediated RNA interfering with the expression of al,3GT,NF- 魏 B in donor heart of mice. Methods: the model of heterotopic heart transplantation from mice to rats was established by modified Heron technique. According to RNA interference donor target gene, there were 8 pairs in each group: control group, empty virus group, RNAi 偽 1t3GT group, RNAi NF- 魏 B group, RNA 偽 1t3GT NF- 魏 B group. The survival time of donor heart was observed, and the expression of ICAM-1, macrophage and NK cells were detected by SABC method. Results: the mean survival time of xenografts in the control group, empty virus group, RNAi 偽 1t3GT group, RNAiNF- 魏 B group and RNAi 偽 1t3GT NF- 魏 B group was (32 鹵2.5) h, (31 鹵2.5) h, (51 鹵4.0) h, (31 鹵1.8) h, (53 鹵7.3) h, respectively. The graft survival time of RNAial,3GT group and RNAial,3GT NF- 魏 B group was significantly longer than that of control group, empty virus group and RNAiNF- 魏 B group (p0.05), and the graft survival time of RNAial,3GT group and RNAial,3GT NF- 魏 B group was significantly longer than that of control group (p0.05), and the graft survival time of RNAial,3GT group and RNAial,3GT NF- 魏 B group was significantly longer than that of control group (p0.05). There was no significant difference in graft survival time between RNAi 偽 1C 3GT group and RNAi 偽 1G 3GT NF- 魏 B group (p0.05). A large amount of SABC was found in the control group, empty virus group and RNAi NF- 魏 B group, and there was no significant difference among the three groups (p0.05). RNAi 偽 1t3GT, RNAi 偽 1t3GT NF- 魏 B group). In the empty virus group, there was no significant difference between the two groups (p0. 05), RNAi 偽 1 + 3GT, RNAi 偽 1 + 3GT NF- 魏 B) (p0. 05). The positive expression of NK cells, macrophages and ICAM-1 was found in the control group, empty virus group and RNAiNF-KB group, and there was no significant difference among the three groups (p0.05). The infiltrating expression of RNAi 偽 1 + 3GT NF- 魏 B group was higher than that of the control group, and the expression of RNAi 偽 1 + 3GT NF- 魏 B group was higher than that of the empty virus group. RNAiNF- 魏 B group increased (p0.05), and RNAi 偽 1t3GT NF- 魏 B group decreased compared with RNAi 偽 1t3GT group (p0.05). Conclusion: xenogeneic antigen Gal 偽 (1t3) Gal plays an important role in xenotransplantation from mice to rats. RNAi can decrease the expression of RNAi and prolong the survival time of graft, decrease the accumulation of complement C3IgNnnnnnnnhus in xenografts, and induce macrophages. The infiltration of natural killer cells increased and the expression of intercellular adhesion molecule-1 increased, which showed the trend of HAR to AVR. Xenograft rejection from mice to rats was characterized by HAR and co-existence with AVR. RNAi NF- 魏 B could reduce the infiltration of graft macrophages and natural killer cells, and decrease the expression of intercellular adhesion molecules, which might reduce the AVR response of xenogeneic cells.
【學位授予單位】：天津醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R392.3

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