摘要
目的:探讨参附注射液对脓毒症模型小鼠脾脏淋巴细胞辅助性T细胞17(T helper cell 17,Th17)及调节性T细胞(regulatory T cell,Treg)免疫平衡的影响。方法:将40只小鼠随机分为假手术组、模型组、参附注射液低剂量组(5 mL·kg^(-1))、参附注射液高剂量组(10 mL·kg^(-1)),每组10只。除假手术组,其余大鼠采用盲肠结扎穿孔手术制备脓毒症模型。造模后2 h各组小鼠每12 h给予腹腔注射相应药物1次,连续4次。计算小鼠脾脏指数;HE染色观察小鼠脾脏病理学改变;TUNEL法检测小鼠脾脏细胞凋亡;流式细胞术检测小鼠脾脏Treg/Th17淋巴细胞百分比;ELISA法检测小鼠血清中白细胞介素-17(interleukin-17,IL-17)、IL-10、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平;Western Blot法检测小鼠脾脏Notch1、Hes1蛋白表达。结果:与模型组比较,参附注射液低、高剂量组小鼠脾脏指数、脾脏细胞凋亡率、脾脏Th17淋巴细胞百分比、脾脏Notch1和Hes1蛋白表达及血清中IL-17、TNF-α水平显著降低(P<0.05),血清IL-10水平、脾脏Treg淋巴细胞百分比水平显著升高(P<0.05),脾脏病理损伤有所改善且呈剂量依赖性。结论:参附注射液可以减轻脓毒症模型小鼠脾脏功能损伤,可能是通过Notch信号通路调节Th17/Treg免疫平衡,从而改善脓毒症的免疫抑制。
Objective:To investigate the effect of Shenfu injection on the immune balance of T helper cell 17(Th17) and regulatory T cell(Treg) in peripheral blood of spleen lymphocytes in sepsis model mice.Methods:40 mice were randomly divided into sham operation group,model group,Shenfu injection low dose group(5 mL·kg^(-1)) and Shenfu injection high dose group(10 mL·kg^(-1)),with 10 mice in each group.In addition to the sham operation group,the sepsis model was prepared by cecal ligation and perforation.Two hours after modeling,mice in each group were intraperitoneally injected with corresponding drugs once every 12 hours for four consecutive times.The spleen organ index of mice was calculated;HE staining was used to observe the histopathological changes of mouse spleen;TUNEL method was used to detect the apoptosis of mouse spleen cells;the percentage of Treg/Th17 lymphocytes in mouse spleen was detected by flow cytometry;interleukin-17(IL-17),IL-10 and tumor necrosis factor-α(TNF-α) in mouse serum were detected by ELISA method;the expression of Notch1 and Hes1 protein in mouse spleen was detected by Western blot.Results:Compared with the model group,the spleen index,spleen cell apoptosis rate,spleen Th17 lymphocyte percentage,spleen Notch1 and Hes1 protein expression,serum IL-17 and TNF-α levels in the low and high dose groups of Shenfu injectionsignificantly increased(P<0.05).The level of serum IL-10,the percentage and level of splenic Treg lymphocytes were significantly reduced(P<0.05),and the pathological damage of the spleen was improved in a dose-dependent manner.Conclusion:Shenfu injection can reduce the damage of spleen function in septic mice,which may regulate Th17/Treg immune balance through Notch signal pathway,so as to improve the immunosuppression of septic mice.
作者
吴峥嵘
李渊
郝素英
WU Zhengrong;LI Yuan;HAO Suying(Dongfang Hospital,Beijing University of Chinese Medcine,Beijing China 100078)
出处
《中医学报》
CAS
2021年第12期2648-2653,共6页
Acta Chinese Medicine
基金
北京中医药大学基本科研业务费项目(青年教师项目)(2017-JYB-JS-196)。
