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目的探讨主要组织相容性复合体(MHC)Ⅱ类基因在多器官功能障碍综合征(MODS)中的表达及意义。方法建立二次打击猪MODS模型。18只雄性家猪随机分为实验组(M组)和正常对照组(C组)每组9只。M组给予失血性休克、再灌注损伤、内毒素血症复合因素；C组仅进行麻醉和动静脉插管。观察7 d后处死动物,取脾组织用Trizol法提取总RNA。设计SLA- DQA(猪MHCⅡ类基因)引物序列,逆转录构建cDNA,用实时荧光定量PCR(Real Time FQ-PCR)检测SLA-DQA mRNA的表达量。UVP计算机图像分析系统绘出标准曲线。结果M组动物死亡率为7/9,有8例发生MODS,发生率为8/9。C组SLA-DQA mRNA拷贝数为(1．376±1．006)×10~3．M组拷贝数为(1．376±1．006)×10~3,两组差异有统计学意义(P＜0．01)。结论MODS模型复制满意。MHCⅡ类基因在MODS中表达下降,可能与逆转录因子(CIITA)的调控和TNF-α、IL-10等多种细胞因子的释放有关。
Objective To investigate the expression of major histocompatibility complex (MHC) Ⅱ gene in multiple organ dysfunction syndrome (MODS) and its significance. Methods A second hit model of pig MODS was established. Eighteen male domestic pigs were randomly divided into experimental group (M group) and normal control group (C group) with 9 in each group. M group was given hemorrhagic shock, reperfusion injury, endotoxemia composite factors; C group only for anesthesia and arteriovenous intubation. After 7 days, the animals were killed and the total RNA was extracted by Trizol method. The primer sequence of SLA-DQA (porcine MHC class II gene) was designed and cDNA was reverse transcribed. The expression of SLA-DQA mRNA was detected by real-time FQ-PCR. UVP computer image analysis system to draw a standard curve. Results The mortality rate of animals in group M was 7/9, and the incidence of MODS in 8 patients was 8/9. The copy number of SLA-DQA mRNA in group C was (1.376 ± 1.006) × 10 ~ 3. The copy number of M group was (1.376 ± 1.006) × 10 ~ 3, the difference between the two groups was statistically significant (P <0.01). Conclusion MODS model is satisfactory. The decreased expression of MHC class II gene in MODS may be related to the regulation of reverse transcription factor (CIITA) and the release of many cytokines such as TNF-α and IL-10.