脓毒症大鼠应激性高血糖与IL-10及CD4^+、CD8^+ T细胞相关性研究

Relationship between stress hyperglycemia and IL-10 and CD4^+ and CD8^+ T cells in rats with sepsis

目的探讨脓毒症大鼠血糖与抗炎-免疫调节因子白细胞介素-10(IL-10)及CD4^+、CD8^+T细胞的相关性。方法 46只成年雄性SD大鼠按随机数字表法分为假手术组(n=18)、脓毒症组(n=28)。采用盲肠结扎穿孔术(CLP)制备大鼠脓毒症模型,制模后脓毒症组分为6、12、18 h 3个亚组。采用ELISA法检测血清IL-10、TNF-α水平;葡萄糖脱氢酶法快速检测大鼠尾静脉血糖,流式细胞仪测定全血中CD4^+、CD8^+T细胞比例;全自动血细胞分析仪检测外周血白细胞计数(WBC)、中性粒细胞绝对值(Neu)、淋巴细胞绝对值(Lym)变化;对不同时间点血糖与IL-10、TNF-α水平及CD4^+、CD8^+T细胞之间进行Pearson直线相关性分析,并进行多元线性回归分析。结果与假手术组相比,脓毒症组制模后各时间点血糖、IL-10、TNF-α、WBC、Neu绝对值均显著升高(P <0. 05); Lym绝对值及CD4^+、CD8^+T细胞比例明显降低(P <0. 05);脓毒症组血糖、Lym绝对值及CD4^+、CD8^+T细胞比例均随制模时间延长降低,IL-10则持续升高(P <0. 05),而TNF-α、WBC、Neu绝对值于12 h升至峰值后逐渐降低(P <0. 05);脓毒症组血糖与IL-10及CD4^+、CD8^+T细胞均存在相关性;进一步多元线性回归分析显示血糖与CD4^+、CD8^+T细胞呈独立相关(P <0. 05)。结论抗炎-免疫调节因子IL-10对脓毒症相关应激性高血糖具有双向调节作用。脓毒症应激性高血糖与T淋巴细胞功能密切相关,在脓毒症应激早期,增高的血糖对免疫细胞有一定的保护作用;随着脓毒症的发展,IL-10参与介导了血糖与免疫细胞的相互抑制。

Objective To investigate the correlation between the blood glucose level and the anti-inflammatory immunoregulatory factors IL-10 and CD4~＋and CD8~＋T cells in septic rats with cecal ligation and puncture. Methods 46 adult male SD rats were randomly divided into sham operation group（n = 18） and sepsis group（n = 28）. The sepsis model was established by cecal ligation and puncture（CLP）. Each group was further divided into three subgroups： 6 h,12 h and 18 h. The serum IL-10 and TNF-α levels were detected by enzyme linked immunosorbent assay（ELISA）; glucose dehydrogenase assay was used to detect the blood glucose level in the tail vein of rats,and the proportion of CD4~＋and CD8~＋T cells in the whole blood was measured by flow cytometry. The changes of blood white blood cell count（WBC）,neutrophil（Neu） absolute value and absolute value of lymphocyte（Lym） were observed by automatic blood cell analyzer,and the changes of blood sugar levels in different time points were related to IL-10,TNF-α,CD4~＋and CD8~＋T cells,and multiple linear regression was carried out. Results The blood sugar,IL-10,TNF-α,WBC and Neu absolute value at each time point of the sepsis group were significantly higher than those in the sham group（P 0. 05）; the proportion of cell absolute value CD4~＋and CD8~＋T cells decreased obviously（P 0. 05）. The proportion of blood sugar,Lym absolute value,CD4~＋and CD8~＋T cells in the sepsis group gradually decreased with the prolongation of mold making time,and the IL-10 level continued to rise（P 0. 05）,while TNF-α,WBC and absolute Neu level were increased to the peak value in 12 h to the peak（P 0. 05）. There was a correlation between blood glucose level and IL-10,CD4~＋and CD8~＋T cells in the sepsis group,and further multivariate linear regression showed that blood glucose was independent of CD4~＋and CD8~＋T cells（P 0. 05）. Conclusion Anti-inflammatory immunoregulatory factor IL-10 has two-way regulation effects on stress related hyperglycemia. In the early stage of sepsis,increased blood sugar has a protective effect on immune cells. With the development of sepsis,IL-10 participates in the mutual inhibition between blood sugar and immune cell function.