Li Hongbing, Li Yuan, Wang Wushi, Peng Zongshuang, Wu Fang
Department of Emergency Medicine, Guiyang First People's Hospital, Guiyang 550002, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2020 May 30;40(5):647-653. doi: 10.12122/j.issn.1673-4254.2020.05.06.
To investigate the role of cholinergic anti-inflammatory pathway (CAP) in neuro-regulation of inflammatory and immune response in the early stage of sepsis.
Sixty-four SD rats were randomly divided into control Group (=8) with normal feeding without any treatment; sham operation group (=8) with laparotomy but without cecal ligation and puncture (CLP), followed by intraperitoneal injection 50 mg/kg piperacillin 3 times a day for 3 consecutive days; and sepsis group (=48) with CLP-induced sepsis. The rat models of sepsis were randomized into model groups (=16) with intraperitoneal injection of piperacillin (50 mg/kg) and normal saline (1 mL/100 g) for 3 times a day for 3 days; GTS-21 group (=16) with additional intraperitoneal injection of 4 mg/kg GTS-21 (once a day for 3 days); and methyllycaconitine (MLA) group (=16) with intraperitoneal injection of MLA (4.8 mg/kg) in addition to piperacillin (once a day for 3 days). Murine Sepsis Score (MSS) of the rats and short-range HRV analysis were recorded. Three days later, the rats were sacrificed and serum levels of TNF-α, IL-1α, IL-10, IL-6, HMGB1, and sCD14 were measured with ELISA. The percentages of CD4CD25 Treg and TH17 lymphocytes and their ratios were measured using flow cytometry.
Compared with the control rats, the septic rats had significantly increased MSS scores and lowered HRV indexes (SDNN, RMSSD, HF, SD1, and SD2; < 0.05); treatment with GTS-21 significantly decreased while MLA increased MSS scores ( < 0.05), but neither of them obviously affected HRV of the rats. Serum levels TNF-α, IL-1α, IL-10, IL-6, HMGB1, and sCD14 and the percentages of CD4CD25 Treg and TH17-positive lymphocytes were significantly higher and Treg/TH17 ratio was significantly lower in the septic rats compared with those in the control group ( < 0.05); treatment with GTS-21 significantly decreased the levels of serum levels of TNF-α, IL-1α, IL-6, HMGB1, and sCD14 and TH17 lymphocyte percentage ( < 0.05), whereas MLA treatment significantly increased serum levels of TNF-α, IL-1α, IL-10, IL-6, HMGB1, and sCD14 and the percentages of CD4 CD25 Treg and TH17-positive lymphocytes and decreased Treg/TH17 ratio in the septic rats ( < 0.05).
CAP plays negative regulatory role in early inflammatory and immune response to sepsis, and some of the HRV indicators can well reflect the regulatory effect of CAP on inflammation and immunity in the septic rats.
探讨胆碱能抗炎通路（CAP）在脓毒症早期炎症和免疫反应神经调节中的作用。
将64只SD大鼠随机分为对照组（n = 8），正常喂养，未作任何处理；假手术组（n = 8），行剖腹术但未进行盲肠结扎和穿刺（CLP），随后腹腔注射50 mg/kg哌拉西林，每天3次，连续3天；脓毒症组（n = 48），采用CLP诱导脓毒症。将脓毒症大鼠模型随机分为模型组（n = 16），腹腔注射哌拉西林（50 mg/kg）和生理盐水（1 mL/100 g），每天3次，共3天；GTS-21组（n = 16），额外腹腔注射4 mg/kg GTS-21（每天1次，共3天）；甲基lycaconitine（MLA）组（n = 16），除哌拉西林外腹腔注射MLA（4.8 mg/kg）（每天1次，共3天）。记录大鼠的小鼠脓毒症评分（MSS）和短程心率变异性分析。3天后，处死大鼠，用ELISA法检测血清中TNF-α、IL-1α、IL-10、IL-6、HMGB1和sCD14的水平。采用流式细胞术检测CD4CD25调节性T细胞（Treg）和辅助性T细胞17（TH17）淋巴细胞的百分比及其比例。
与对照大鼠相比，脓毒症大鼠的MSS评分显著升高，心率变异性指标降低（SDNN、RMSSD、HF、SD1和SD2；P < 0.05）；GTS-21治疗显著降低而MLA增加MSS评分（P < 0.05），但两者均未明显影响大鼠的心率变异性。与对照组相比，脓毒症大鼠血清中TNF-α、IL-1α、IL-10、IL-6、HMGB1和sCD14水平以及CD4CD25 Treg和TH17阳性淋巴细胞百分比显著升高，Treg/TH17比值显著降低（P < 0.05）；GTS-21治疗显著降低血清中TNF-α、IL-1α、IL-6、HMGB1和sCD14水平以及TH17淋巴细胞百分比（P < 0.05），而MLA治疗显著增加脓毒症大鼠血清中TNF-α、IL-1α、IL-10、IL-6、HMGB1和sCD14水平以及CD4 CD25 Treg和TH17阳性淋巴细胞百分比，并降低Treg/TH17比值（P < 0.05）。
CAP在脓毒症早期炎症和免疫反应中起负性调节作用，部分心率变异性指标可较好反映CAP对脓毒症大鼠炎症和免疫的调节作用。
