氧化苦参碱通过影响辅助性T细胞17/调节性T细胞的比例对急性淋巴细胞白血病小鼠免疫功能的影响

Effect of oxymatrine on immune function of acute lymphoblastic leukemia mice by influencing Th17/Treg ratio

目的探讨氧化苦参碱(OMT)通过影响辅助性T细胞17(Th17)/调节性T细胞(Treg)的比例对急性淋巴细胞白血病小鼠免疫功能的影响。方法2021年6—12月将50只小鼠按随机数字表法分为正常对照组、非干预组、OMT低、中、高剂量组,正常对照组灌胃等量生理盐水,其余四组小鼠于右前侧皮下注射L1210细胞悬浮液建立急性淋巴细胞白血病(ALL)小鼠模型。造模成功后进行干预治疗,OMT低、中、高剂量组分别灌胃50、100、200 mg/kg剂量的OMT,正常对照组和非干预组注射灌胃生理盐水。采用人工计数法进行白细胞计数及分类;计算脾脏和胸腺指数;ELISA法检测小鼠血清中白细胞介素(IL)-17、IL-22、转化生长因子-β(TGF-β)和IL-10水平;流式细胞术检测CD4^(+)、CD8^(+)百分比、CD4^(+)/CD8^(+)及Th17、Treg比例。结果OMT可增加小鼠体质量、中性粒细胞数量、CD4^(+)/CD8^(+)、脾脏和胸腺指数,降低皮下瘤质量、L1210细胞、淋巴细胞、白细胞数量及Th17/Treg。正常对照组、非干预组、OMT低、中、高剂量组IL-17水平分别为(70.81±6.24)ng/L、(257.34±22.93)ng/L、(189.36±18.54)ng/L、(124.47±12.73)ng/L、(81.13±7.39)ng/L;正常对照组、非干预组、OMT低、中、高剂量组IL-22水平分别为(66.29±6.85)ng/L、(245.08±22.93)ng/L、(153.43±17.29)ng/L、(101.30±11.33)ng/L、(77.87±6.74)ng/L;正常对照组、非干预组、OMT低、中、高剂量组TGF-β水平分别为(281.94±25.91)ng/L、(71.01±7.25)ng/L、(90.39±10.83)ng/L、(142.00±16.00)ng/L、(221.66±19.59)ng/L;正常对照组、非干预组、OMT低、中、高剂量组IL-10水平分别为(157.68±13.34)ng/L、(42.74±4.80)ng/L、(58.23±6.32)ng/L、(80.14±8.73)ng/L、(96.27±10.87)ng/L。结论OMT能够提高ALL小鼠免疫功能,其机制可能与改善Th17/Treg细胞平衡有关。

Objective To explore the effect of oxymatrine(OMT)on immune function of acute lymphoblastic leukemia mice by affecting the ratio of T-helper cell 17(Th17)/regulatory-T cell(Treg).Methods From June 2021 to December,2021,50 mice were randomly assigned into normal control group,non-intervention group,low-,medium-,and high-dose OMT groups according to random number table method.Normal control group was given equal amount of intragastric normal saline,and the remaining four groups were subcutaneously injected with L1210 cell suspension in the right anterior side to establish acute lymphoblastic leukemia(ALL)mice model.After successful modeling,intervention was performed.The low-,medium-,and high-dose OMT groups were given 50,100 and 200 mg/kg of OMT,respectively.The mice of normal control group and non-intervention group were given normal saline.White blood cell count and classification were carried out by manual counting method.Spleen and thymus indexes were calculated.Serum levels of interleukin(IL)-17,IL-22,transforming growth factor-β(TGF-β)and interleukin-10(IL-10)were detected by the enzyme linked immunosorbent assay(ELISA).The percentages of CD4^(+),CD8^(+),CD4^(+)/CD8^(+),and Th17/Treg ratio were detected by flow cytometry.Results OMT could increase the body mass,neutrophil count,CD4^(+)/CD8^(+),spleen and thymus index,and decrease the subcutaneous tumor mass,the counts of L1210 cells,lymphocytes and leukocytes,and the Th17/Treg ratio.The levels of IL-17,IL-22,TGF-β,IL-10 in normal control group,non-intervention group,low-,medium-and high-dose OMT groups were[(70.81±6.24)ng/L,(257.34±22.93)ng/L,(189.36±18.54)ng/L,(124.47±12.73)ng/L and(81.13±7.39)ng/L],[(66.29±6.85)ng/L,(245.08±22.93)ng/L,(153.43±17.29)ng/L,(101.30±11.33)ng/L and(77.87±6.74)ng/L],[(281.94±25.91)ng/L,(71.01±7.25)ng/L,(90.39±10.83)ng/L,(142.00±16.00)ng/L and(221.66±19.59)ng/L],[(157.68±13.34)ng/L,(42.74±4.80)ng/L,(58.23±6.32)ng/L,(80.14±8.73)ng/L and(96.27±10.87)ng/L],respectively.Conclusion OMT can improve the immune function of ALL mice,whose mechanism may be related to the improvement in the balance of Th17 versus Treg cells.