摘要
目的肺癌根治术围术期使用右美托咪定可有效提高机体免疫功能,但是其具体作用机制尚不明确。本研究基于白细胞介素-6(interleukin 6,IL-6)/STAT3通路分析右美托咪定对Lewis肺癌小鼠免疫功能的影响,并探讨右美托咪定改善Lewis肺癌患者免疫功能的分子机制。方法 60只雄性C57BL/6小鼠按随机数字表法分为对照组、模型组、右美托咪定(30μg/kg)组、SH-4-54(IL-6/STAT3通路抑制剂,10mg/kg)组和顺铂(DDP,2mg/kg)组,右腋下注射Lewis肺癌细胞建立Lewis肺癌小鼠模型。给药14d后处死小鼠,称取体质量及瘤体、胸腺、脾质量,计算各组小鼠抑瘤率、胸腺指数和脾指数;采用流式细胞术检测外周血中T淋巴细胞CD4^+和CD8^+百分比,并计算其比值;采用酶联免疫吸附法检测血清中IL-2、IL-10、IL-6和γ-干扰素(interferon-γ,IFN-γ)的含量;蛋白免疫印迹法检测IL-6表达及p-STAT3/STAT3。采用SPSS 20.0对数据进行统计分析,多组间均值比较采用单因素方差分析,组间两两多重比较采用SNK-q或LSD-t检验;不符合正态分布采用秩转换的Kruskal-Wallis H检验。结果 (1)小鼠瘤体质量比较:对照组、模型组、右美托咪定组、SH-4-54组和DDP组瘤体质量(g)分别为0、0.72±0.03、0.57±0.18、0.56±0.16和0.31±0.11,5组小鼠瘤体质量差异有统计学意义,F=67.580,P<0.001。两两多重比较显示,右美托咪定组、SH-4-54组及DDP组瘤体质量低于模型组,P值分别为0.026、0.015和<0.001;DDP组瘤体质量低于右美托咪定组和SH-4-54组,均P<0.001。(2)小鼠胸腺指数比较:对照组、模型组、右美托咪定组、SH-4-54组和DDP组胸腺指数(mg/g)分别为2.45±0.19、1.42±0.20、2.15±0.17、2.11±0.18和0.96±0.17,5组胸腺指数差异均有统计学意义,F=134.349,P<0.001。两两多重比较显示,模型组小鼠胸腺指数低于对照组,均P<0.001;右美托咪定组、SH-4-54组小鼠胸腺指数高于模型组,DDP组小鼠脾指数、胸腺指数低于模型组,均P<0.001。(3)小鼠外周血CD4^+/CD8^+结果比较:对照组、模型组、右美托咪定组、SH-4-54组和DDP组CD4^+/CD8^+分别为1.46±0.09、0.96±0.09、1.28±0.08、1.30±0.09和0.70±0.08,5组小鼠外周血CD4^+/CD8^+差异均有统计学意义,F=151.051,P<0.001。两两多重比较显示,模型组小鼠CD4^+/CD8^+比值低于对照组,P<0.001;右美托咪定组、SH-4-54组小鼠CD4^+/CD8^+比值高于模型组,均P<0.001;DDP组小鼠CD4^+/CD8^+比值低于模型组,均P<0.001。(4)小鼠血清IL-2、IFN-γ、IL-10和IL-6水平比较:对照组、模型组、右美托咪定组、SH-4-54组和DDP组IL-2分别为139.21±21.86、73.97±20.05、106.36±19.01、103.91±20.03和50.76±23.38,IFN-γ分别为109.76±10.06、56.06±10.97、86.82±11.64、85.98±11.76和40.03±9.39,IL-6分别为82.36±13.32、228.18±15.38、134.27±12.37、136.43±13.05和251.95±13.26;IL-10分别为97.41±17.39、240.01±20.26、156.38±22.41、157.41±20.36和265.73±22.35,5组小鼠外周血IL-2、IFN-γ、IL-10和IL-6差异均有统计学意义,F值分别为31.262、78.341、330.658和132.326,均P<0.001。两两多重比较显示,模型组小鼠IL-2、IFN-γ低于对照组,IL-10和IL-6高于对照组,P<0.001;右美托咪定组、SH-4-54组小鼠IL-2、IFN-γ高于模型组,IL-10和IL-6低于模型组,均P<0.001;DDP组小鼠IL-2、IFN-γ低于模型组,IL-10和IL-6高于模型组,均P<0.001。(5)小鼠IL-6蛋白和p-STAT3/STAT3的测定结果比较:对照组、模型组、右美托咪定组、SH-4-54组和DDP组IL-6蛋白表达分别为0.41±0.08、0.97±0.09、0.69±0.08、0.96±0.08和1.36±0.10,p-STAT3/STAT3蛋白表达比值分别为0.76±0.11、1.42±0.12、0.58±0.13、0.53±0.14和2.32±0.14,5组IL-6和p-STAT3/STAT3水平差异均有统计学意义,F值分别为201.828和417.545,均P<0.001。两两多重比较显示,模型组小鼠IL-6水平和p-STAT3/STAT3高于对照组,均P<0.001;右美托咪定组小鼠IL-6水平和p-STAT3/STAT3低于模型组,均P<0.001;SH-4-54组小鼠p-STAT3/STAT3低于模型组,P<0.001;DDP组小鼠IL-6蛋白水平、p-STAT3/STAT3高于模型组,均P<0.001;右美托咪定组、SH-4-54组小鼠IL-6蛋白水平和p-STAT3/STAT3低于DDP组,均P<0.001。结论右美托咪定可以改善Lewis肺癌小鼠的免疫功能,其机制可能是通过抑制IL-6/STAT3信号通路实现的。
OBJECTIVE The use of dexmedetomidine in the perioperative period of lung cancer radical surgery can effectively improve the immune function of the body,but the specific mechanism is not clear.Therefore,this study was based on IL-6/STAT3 pathway to explore the effect of dexmedetomidine on the immune function of Lewis lung cancer mice,and to elucidated the molecular mechanism of dexmedetomidine improving the immune function of Lewis lung cancer patients.METHODS Totally 60 male C57 BL/6 mice were divided into control group,model group,dexmedetomidine(30μg/kg),SH-4-54(IL-6/STAT3 pathway inhibitor,10 mg/kg)group and cisplatin(DDP,2 mg/kg)group,Lewis lung cancer model was established by injecting Lewis lung cancer cells into right armpit.The mice were killed 14 days after administration,and the weight of tumor body,thymus and spleen were weighed;the percentage of CD4^+and CD8^+in peripheral blood was detected by flow cytometry,and the ratio was calculated;the contents of interleukin 2(IL-2),IL-10,IL-6 and interferon-γ(IFN-γ)in serum were detected by enzyme linked immunosorbent assay(ELISA);the expression of IL-6 and p-STAT3/STAT3 were detected by Western blot.SPSS 20.0 was used for statistical analysis of data.,single factor analysis of variance was used for mean comparison among multiple groups,SNK-q or LSD-t test was used for the comparison between two groups.Kruskal-Wallis Htest was used for rank transformation for non-conforming normal distribution,the test level wasα= 0.05(double tail).RESULTS (1)Comparison of tumor mass in mice:The tumor mass(g)of the control group,the model group,the dexmedetomidine group,the SH-4-54 group and the cisplatin group were 0,0.72±0.03,0.57±0.18,0.56±0.16 and 0.31±0.11,respectively;single factor analysis of variance showed that there was significant difference in tumor mass among the five groups(F=67.580,P<0.001);the double comparison showed that the tumor mass of dexmedetomidine group,SH-4-54 group and cisplatin group was lower than that of the model group(Pvalues were 0.026,0.015 and<0.001,respectively);the tumor mass of cisplatin group was lower than that of dexmedetomidine group and SH-4-54 group(all P<0.001).(2)Comparison of thymus index in mice:The thymus index(mg/g)of control group,model group,dexmedetomidine group,SH-4-54 group and cisplatin group were 2.45±0.19,1.42±0.20,2.15±0.17,2.11±0.18 and 0.96±0.17,respectively;single factor analysis of variance showed that the thymus index of the five groups had statistical significance(F=134.349,P<0.001);the double comparison showed that the thymus index of the model group was lower than that of the control group(all P<0.001);the thymus index of the right metomidine group and the SH-4-54 group was higher than that of the model group;the spleen index and thymus index of the cisplatin group were lower than that of the model group(all P<0.001).(3)Comparison of CD4^+/CD8^+results in peripheral blood of mice:The CD4^+/CD8^+in the control group,the model group,the dexmedetomidine group,the SH-4-54 group and the cisplatin group were 1.46±0.09,0.96±0.09,1.28±0.08,1.30±0.09 and 0.70±0.08,respectively;single factor analysis of variance showed that the difference of CD4^+/CD8^+in peripheral blood of the five groups was statistically significant(F=151.051,P<0.001);the double comparison showed that the ratio of CD4^+/CD8^+in model group was lower than that in control group(P<0.001);the ratio of CD4^+/CD8^+in dexmedetomidine group and SH-4-54 group was higher than that in model group(all P<0.001),and cisplatin group was lower than that in model group(all P<0.001).(4)Comparison of serum IL-2,IFN-γ,IL-10 and IL-6 levels in mice:The levels of IL-2 in the control group,model group,dexmedetomidine group,sh-4-54 group and cisplatin group were 139.21±21.86,73.97±20.05,106.36±19.01,103.91±20.03 and 50.76±23.38,respectively;IFN-γwere109.76±10.06,56.06±10.97,86.82±11.64,85.98±11.76,40.03±9.39;IL-6 were 82.36±13.32,228.18±15.38,134.27±12.37,136.43±13.05,251.95±13.26;IL-10 were 97.41±17.39,240.01±20.26,156.38±22.41,157.41±20.36,265.73±22.35;single factor analysis of variance showed that the differences of IL-2,IFN-γ,IL-10 and IL-6 were statistically significant(F=31.262,78.341,330.658,132.326,P<0.001);the double comparison showed that IL-2 and IFN-γin the model group were lower than those in the control group,IL-10 and IL-6 were higher than those in the control group(P<0.001);IL-2 and IFN-γin the right metomidine group and sh-4-54 group were higher than those in the model group,IL-10 and IL-6 were lower than those in the model group(all P<0.001),IL-2 and IFN-γin the cisplatin group were lower than those in the model group,IL-10 and IL-6 were higher than those in the model group(all P<0.001).(5)Comparison of IL-6 protein and p-STAT3/STAT3 in mice:The expression of IL-6 protein in the control group,model group,dextromethoridine group,SH-4-54 group and cisplatin group were 0.41±0.08,0.97±0.09,0.69±0.08,0.96±0.08 and 1.36±0.10,respectively;the expression ratio of p-STAT3/STAT3 protein were 0.76±0.11,1.42±0.12,0.58±0.13,0.53±0.14 and 2.32±0.14,respectively;single factor analysis of variance showed that there were significant differences in IL-6 protein and p-STAT 3/STAT 3 levels among the five groups(Fvalues were 201.828 and417.545,respectively(all P<0.001);the double comparison showed that the levels of IL-6 protein and p-STAT 3/STAT 3 in model group were higher than those in control group(all P<0.001);the levels of IL-6 protein and p-STAT 3/STAT 3 in the dexmedetomidine group were lower than those in the model group(P<0.001),and p-STAT 3/STAT 3 in the SH-4-54 group were lower than those in the model group(P<0.001),the levels of IL-6 protein and p-STAT 3/STAT 3 in cisplatin group were higher than those in model group(all P<0.001),while the levels of IL-6 protein and p-STAT 3/STAT 3 in dexmedetomidine group and SH-4-54 group were lower than those in cisplatin group(all P<0.001).CONCLUSION Dexmedetomidine can improve the immune function of Lewis lung cancer mice,possibly by inhibiting the IL-6/STAT 3 signaling pathway.
作者
苏晓英
王红又
董倩
韩建军
SU Xiao-ying;WANG Hcmg-you;DONG Qian;HAN Jian-jun(Sichuan Mental Health Center,Mianyang Third People's Hospital,Mianyang 621000,P.R.China)
出处
《中华肿瘤防治杂志》
CAS
北大核心
2020年第13期1040-1048,共9页
Chinese Journal of Cancer Prevention and Treatment
