黄芪-莪术对C5a介导JAK2/STAT3通路调控Lewis肺癌小鼠Th17/Treg细胞平衡影响的实验研究  

Astragali Radix-Curcumae Rhizoma Regulates Th17/Treg Cell Balance in the Mouse Model of Lewis Lung Carcinoma via C5a-Mediated JAK2/STAT3 Pathway

作　　者：田培裕 于泓洋 李潇[1,2] 闫梓乔 窦永起 TIAN Pei-yu;YU Hong-yang;LI Xiao;YAN Zi-qiao;DOU Yong-qi(Chinese PLA Medical School,Beijing 100853;Unit 71901 of the People′s Liberation Army,Liaocheng Shandong 252000;Department of Traditional Chinese Medicine,the Sixth Medical Center,Chinese PLA General Hospital,Beijing 100089)

机构地区：[1]中国人民解放军医学院,北京100853 [2]71901部队,山东聊城252000 [3]中国人民解放军总医院第六医学中心中医医学部,北京100089

出　　处：《世界中西医结合杂志》2024年第3期425-432,共8页World Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(82174118)。

摘　　要：目的探讨黄芪-莪术影响C5a介导JAK2/STAT3通路调控肿瘤微环境中Th17/Treg细胞平衡,从而阐明其在此途径抑制肿瘤进展的相关机制。方法40只雄性C57BL/6小鼠按随机数字表法分为空白对照组、模型对照组、中药干预组、阳性对照药(PMX53)组,每组各10只。腋下接种Lewis细胞建立肺癌小鼠模型,于接种后第3天开始,中药干预组按8.2 g/kg剂量给予中药浓煎液灌胃,模型对照组和PMX53组予等容积灭菌双蒸水灌胃,连续14 d;PMX53组分别于第3、6、9、12、15天按1mg/kg剂量给予腹腔注射PMX53,模型对照组和中药干预组同时腹腔注射等容PBS溶液。每2 d测算各组小鼠肿瘤体积、绘制肿瘤生长曲线;于第15天处死,剖取瘤块,ELISA法检测血清C5a、IL6、IL-10、IL-17、TGF-β等因子水平,流式细胞术检测并计算外周血和肿瘤组织中Th17/Treg细胞比例,Western Blot和Real-time PCR法检测肿瘤组织中JAK2/STAT3信号通路相关蛋白及mRNA表达。结果与模型对照组比较,中药干预组和PMX53组肿瘤生长较缓慢,补体C5a、JAK2和STAT3蛋白及mRNA水平、p-JAK2/JAK2、p-STAT3/STAT3比值、Treg占比均降低,SOCS3蛋白及mRNA水平、Th17/Treg比例增高,差异有统计学意义(P<0.05)。结论黄芪-莪术可降低补体C5a进而抑制JAK2/STAT3通路活化,并调节肿瘤微环境中Th17/Treg平衡达到抑制肿瘤生长的作用。Objective To study the regulatory effect of Astragali Radix-Curcumae Rhizoma(AR-CR)on Th17/Treg cell balance in the tumor microenvironment by regulating the C5a-mediated Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)pathway,thereby deciphering the mechanism of this herb pair in inhibiting tumor progression.Methods Forty male C57BL/6 mice were assigned by the random number table method into blank control,model control,AR-CR,and positive control(PMX53)groups,with 10 mice in each group.Lewis cells were inoculated in the armpit to establish a mouse model of lung cancer.From day 3 after inoculation,the mice in the AR-CR group were administrated with the decoction of AR-CR at a dose of 8.2 g/kg by gavage,and those in the model control and PMX53 groups were administrated with the equal volume of sterilized double-distilled water by gavage for 14 consecutive days.The PMX53 group was subjected to intraperitoneal injection of PMX53 at a dose of 1 mg/kg on days 3,6,9,12,and 15,respectively,and the model control and AR-CR groups were injected intraperitoneally with equal-volume PBS solution at the same time.The tumor volume of each group was measured every 2 days and the tumor growth curve was drawn.All the mice were executed on day 15,and the tumor mass was collected.The serum levels of C5a,interleukin(IL)-6,IL-10,IL-17,and transforming growth factor(TGF)-βwere determined by ELISA.The proportion of Th17/Treg cells in the peripheral blood and tumor tissue was examined by flow cytometry and calculated.The protein and mRNA levels of JAK2/STAT3 signaling pathway-related factors were determined by Western blotting and real-time PCR,respectively.Results Compared with the model control group,the AR-CR and PMX53 groups showed slow tumor growth,down-regulated mRNA and protein levels of C5a,JAK2,and STAT3,decreased p-JAK2/JAK2 and p-STAT3/STAT3 ratios,reduced proportion of Treg cells,up-regulated protein and mRNA levels of SOCS3,and increased Th17/Treg ratio(all P<0.05).Conclusion AR-CR can reduce com

关 键 词：肿瘤微环境 补体C5A JAK2/STAT3通路 TH17/TREG 黄芪-莪术 

分 类 号：R285.5[医药卫生—中药学]