四君子汤改善C57小鼠恶病质状态肌肉消耗的疗效及机制研究  

Study on the efficacy and mechanism of Sijunzi Decoction in improving cachexia muscle consumption in C57 mice

作　　者：朱健 吉兆奕[2] 焦拥政[1] 朱晓云[3] 曹 袁国兴 吴洁[2] ZHU Jian;JI Zhaoyi;JIAO Yongzheng;ZHU Xiaoyun;CAO Yan;YUAN Guoxing;WU Jie(Guang'anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053,China)

机构地区：[1]中国中医科学院广安门医院男科,北京100053 [2]中国中医科学院广安门医院血液科,北京100053 [3]中国中医科学院广安门医院糖尿病分子实验室,北京100053 [4]北京市东城区第一人民医院中西医结合科

出　　处：《环球中医药》2022年第5期752-758,共7页Global Traditional Chinese Medicine

基　　金：国家自然科学基金(81373596);北京市自然科学基金(7142141)。

摘　　要：目的观察四君子汤缓解或改善C57小鼠恶病质状态肌肉消耗的疗效及机制。方法C57BL/6小鼠皮下注射lewis肺腺癌肿瘤细胞建立肿瘤恶病质模型,按照随机数字表法分为正常组、模型组、四君子组1 g/(mL·d)和地屈孕酮组予甲地孕酮溶液0.024 g/(kg·d),每组6只,连续给药14天。检测各组小鼠的体质量、肿瘤大小、脾/肝脏质量、骨骼肌质量,电镜下观察骨骼肌纤维形态的变化,酶联免疫吸附分析(enzyme linked immunosorbent assay,ELISA)法检测血清炎症因子肿瘤坏死因子(tumor necrosis factor,TNF-α)、激活素-A(Activin-A)水平,蛋白免疫印迹(western blot,WB)法及实时荧光定量PCR(real-time quantitative PCR,qRT-PCR)检测肌萎缩基因Myostatin、Activin-A、MuRF-1表达水平。结果(1)与模型组相比,四君子组的瘤体质量降低,肝脏质量降低,骨骼肌绝对质量增加(P<0.05)。(2)与正常组相比,模型组小鼠腓肠肌肌细胞排列疏松,肌纤维细胞缩小,细胞间隙增大,肌纤维细胞大小差异较大。与模型组相比,四君子组小鼠腓肠肌肌细胞排列相对紧密,细胞间隙缩小。(3)与正常组相比,模型组和四君子组小鼠血清中TNF-α、Activin-A水平明显升高(P<0.05);与模型组相比,地屈孕酮组和四君子组Activin-A呈低表达(P<0.05)。(4)qRT-PCR检测显示,与模型组相比,四君子组Myostatin、MuRF1的mRNA表达明显降低(P<0.05);Western blot结果显示四君子汤能够降低肌萎缩相关蛋白Activin-A的表达,结果与mRNA表达结果一致。结论四君子汤对荷瘤小鼠恶病质状态下的肌肉消耗有明显的缓解或改善作用,可能通过降低血清中Activin-A表达水平,改善肝肿大以及骨骼肌萎缩症状,降低Myostatin、MuRF1的mRNA表达,调节Myostatin/Actvin-ActRIIB信号途径抑制肺癌恶病质小鼠骨骼肌萎缩。Objective To establish a lung cancer cachexia model in C57 mice to observe the efficacy and mechanism of Sijunzi Decoction in alleviating and improving muscle wasting in C57 mice cachexia.Methods C57BL/6 mice were injected subcutaneously with Lewis lung adenocarcinoma tumor cells to establish thetumor cachexia model.According to the random number table method,they were divided into 4 groups:the normal group,the model group,the dydrogesterone group and the Sijunzi Decoction group,6 mice in each group,continuously given medicine for 14 days.The weight,tumor size,spleen/liver weight,skeletal muscle weight,influence on skeletal muscle fiber morphology,cachexia muscle wasting and the expression of key genes in muscle atrophy were examined.Results(1)Compared with the model group,the tumor diameter in the Sijunzi group was reduced,the liver mass in the Sijunzi Decoction group was reduced,and the gastrocnemius absolute weight in the Sijunzi group was increased(P<0.05).(2)Compared with the normal group,the gastrocnemius muscle cells of cachexia mice are loosely arranged,the muscle fiber cells are reduced,the intercellular space is enlarged,and the size of the muscle fiber cells is different.Compared with the model group,the gastrocnemius muscle cells of the Sijunzi Decoction group were arranged relatively tightly,and the intercellular space was narrowed.(3)Compared with the normal group,the serum levels of TNF-a and Activin-A in the model group and the Sijunzi group were significantly increased(P<0.05).Compared with the model group,the dydrogesterone group and the Sijunzi group Activin-A showed low expression(P<0.05).(4)Real-time quantitative PCR(qRT-PCR)detection showed that compared with the model group,Sijunzi Decoction could significantly reduce the mRNA expression of Myostatin and MuRF1(P<0.05).Western blot results showed that Sijunzi Decoction could reduce the expression of muscle wasting-related protein Activin-A,and the result is consistent with the mRNA expression result.Conclusion Sijunzi Decoction can significa

关 键 词：C57BL/6小鼠 lewis细胞 肿瘤恶病质 骨骼肌萎缩 Myostatin/Actvin-ActRIIB信号通路 四君子汤 

分 类 号：R285.5[医药卫生—中药学]