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Title: [Influence of vagus nerve on multiple organ function and immune reaction of T lymphocytes in septic rats].
Author: Ren C, Li XH, Wu Y, Dong N, Yao YM.
Journal: Zhonghua Shao Shang Za Zhi; 2018 Nov 20; 34(11):815-820. PubMed ID: 30481924.
Abstract:
Objective: To explore influence of vagus nerve on multiple organ function and immune reaction of T lymphocytes in septic rats. Methods: Forty Sprague-Dawley rats were divided into sham injury group, sepsis group, vagus nerve stimulation (VNS) group, and vagotomy (VGX) group, according to the random number table, with 10 rats in each group. Rats in sepsis group, VNS group, and VGX group were inflicted with sepsis by cecal ligation and puncture (CLP). Rats in VNS group were given electrical stimulation on left cervical vagus nerve for 15 min right after CLP. Rats in VGX group underwent vagotomy of left cervical vagus nerve at 30 min before CLP. At 24 h after CLP, serum of rats was collected to detect levels of alanine transaminase (ALT), aspartate aminotransferase (AST), glycocholic acid (CG), creatine kinase (CK), myocardial creatine kinase (CK-MB), blood urea nitrogen (BUN), and serum creatinine by fully automatic biochemistry analyzer. The left lung of rats was collected to determine wet or dry mass, and wet to dry (W/D) ratio was calculated. The right lung of rats was collected to measure the activity of pulmonary myeloperoxidase (MPO) by enzyme linked immunosorbent assay (ELISA). Spleen of rats was collected to determine the proliferative activity of CD4(+) T lymphocytes by cell counting kit 8, and real-time fluorescence quantitative polymerase chain reaction and ELISA were used to quantify mRNA expressions and levels of interleukin 2 (IL-2), interferon-γ, and IL-4, respectively. Data were processed with one-way analysis of variance and Tukey's honest significant difference test. Results: (1) The levels of serum ALT, AST, CG, CK, CK-MB, BUN, and creatinine, pulmonary W/D ratio, as well as MPO activity of rats in sepsis group were significantly higher than those in sham injury group and VNS group (P<0.01) and were significantly lower than those in VGX group (P<0.01). (2) The proliferative activity of CD4(+) T lymphocytes of rats in sepsis group was 0.93±0.03, which was significantly lower than 1.54±0.07 of rats in sham injury group (P<0.01). The proliferative activity of CD4(+) T lymphocytes of rats in VNS group was 1.15±0.15, which was significantly higher than that of rats in sepsis group (P<0.01). The proliferative activity of CD4(+) T lymphocytes of rats in VGX group was 0.75±0.06, which was obviously lower than that of rats in sepsis group (P<0.01). (3) In comparison with those of rats in sham injury group, the levels of IL-2 and interferon-γ in CD4(+) T lymphocytes of rats in sepsis group were markedly decreased (P<0.01), while the level of IL-4 was significantly increased (P<0.01). In comparison with those of rats in sepsis group, the levels of IL-2 and interferon-γ in CD4(+) T lymphocytes of rats in VNS group were obviously increased (P<0.01), while the level of IL-4 was markedly decreased (P<0.01). As compared with those of rats in sepsis group, the levels of IL-2 and interferon-γ in CD4(+) T lymphocytes of rats in VGX group were markedly decreased (P<0.01), while the level of IL-4 was significantly increased (P<0.01). (4) As compared with those of rats in sham injury group, expressions of IL-2 and interferon-γ mRNA in CD4(+) T lymphocytes of rats in sepsis group were markedly decreased (P<0.01), while expression of IL-4 mRNA was significantly increased (P<0.01). Expressions of IL-2 and interferon-γ mRNA in CD4(+) T lymphocytes of rats in VNS group were obviously increased when compared with those of rats in sepsis group (P<0.01), while expression of IL-4 mRNA was markedly decreased (P<0.01). In comparison with those of rats in sepsis group, expressions of IL-2 and interferon-γ mRNA in CD4(+) T lymphocytes of rats in VGX group were markedly decreased (P<0.01), while expression of IL-4 mRNA was significantly increased (P<0.01). Conclusions: Electrical stimulation of vagus nerve can significantly improve multiple organ dysfunction and reverse immunosuppression of T lymphocytes in septic rats, while vagotomy of vagus nerve may enhance the susceptibility of rats to sepsis. 目的： 探讨迷走神经对脓毒症大鼠多器官功能及T淋巴细胞免疫反应的影响。 方法： 将40只SD大鼠按随机数字表法分成假伤组、脓毒症组、迷走神经刺激(VNS)组、迷走神经离断(VGX)组，每组各10只。脓毒症组、VNS组及VGX组大鼠采用盲肠结扎穿孔术(CLP)构建脓毒症模型，VNS组大鼠在CLP后立即予左侧颈部迷走神经电刺激15 min，VGX组大鼠于CLP前30 min行左侧颈部迷走神经离断术。CLP后24 h收集大鼠血清，全自动生化分析仪检测丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、甘胆酸、肌酸激酶、心肌型肌酸激酶同工酶(CK－MB)、血尿素氮、血肌酐水平。取大鼠左肺，称湿、干质量，计算肺湿干质量比；取大鼠右肺，酶联免疫吸附测定(ELISA)法检测肺组织髓过氧化物酶(MPO)活性。取大鼠脾脏，采用细胞计数试剂盒8法检测CD4(＋)T淋巴细胞增殖活性，ELISA法及实时荧光定量PCR法检测白细胞介素2(IL－2)、γ干扰素及IL－4水平及其mRNA表达。对数据行单因素方差分析及Tukey′s HSD检验。 结果： (1)脓毒症组大鼠血清ALT、AST、甘胆酸、肌酸激酶、CK－MB、血尿素氮、血肌酐水平与肺湿干质量比及肺组织MPO活性明显高于假伤组和VNS组(P<0.01)，明显低于VGX组(P<0.01)。(2)脓毒症组大鼠CD4(＋)T淋巴细胞增殖活性为0.93±0.03，明显低于假伤组的1.54±0.07(P<0.01)。VNS组大鼠CD4(＋)T淋巴细胞增殖活性为1.15±0.15，明显高于脓毒症组(P<0.01)。VGX组大鼠CD4(＋)T淋巴细胞增殖活性为0.75±0.06，明显低于脓毒症组(P<0.01)。(3)与假伤组比较，脓毒症组大鼠CD4(＋)T淋巴细胞IL－2及γ干扰素水平明显下降(P<0.01)，IL－4水平明显升高(P<0.01)。与脓毒症组比较，VNS组大鼠CD4(＋)T淋巴细胞IL－2及γ干扰素水平明显升高(P<0.01)，而IL－4水平明显降低(P<0.01)。与脓毒症组比较，VGX组大鼠CD4(＋)T淋巴细胞IL－2及γ干扰素水平明显降低(P<0.01)，IL－4水平明显升高(P<0.01)。(4)与假伤组比较，脓毒症组大鼠CD4(＋) T淋巴细胞IL－2及γ干扰素的mRNA表达明显降低(P<0.01)，而IL－4 mRNA表达明显升高(P<0.01)。与脓毒症组比较，VNS组大鼠CD4(＋)T淋巴细胞IL－2及γ干扰素的mRNA表达明显升高(P<0.01)，IL－4 mRNA表达明显降低(P<0.01)。与脓毒症组比较，VGX组大鼠CD4(＋)T淋巴细胞IL－2及γ干扰素的mRNA表达明显降低(P<0.01)，IL－4 mRNA表达明显升高(P<0.01)。 结论： 电刺激迷走神经能显著改善脓毒症大鼠多器官功能障碍并逆转T淋巴细胞免疫功能抑制，离断迷走神经则可导致大鼠对脓毒症的易感性增加。.

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