骨痛灵方对肺癌骨转移疼痛模型小鼠RANKL/OPG的影响  

The effect of Gu-Tong-Ling prescription on the expressings of inflammatory factors and RANKL/OPG in lung cancer bone metastasis mouse mode

作　　者：罗琴琴 徐振晔[2] 邓海滨[2] LUO Qin-qin;XU Zhen-ye;DENG Hai-bin(Wuhan Hospital of Traditional Chinese and Western Medicine Affiliated to Tongji Hospital of Huazhong University of Science and Technology,Chinese Medicine Department,Wuhan Hubei 430000;Longhua Hospital Affiliated to Shanghai Univercity of Traditional Medicine,Oncology Department,Shanghai 200032)

机构地区：[1]华中科技大学同济医学院附属武汉市中西医结合医院中医科,湖北武汉430000 [2]上海中医药大学附属龙华医院肿瘤科,上海200032

出　　处：《世界中西医结合杂志》2020年第6期1067-1071,1083,共6页World Journal of Integrated Traditional and Western Medicine

基　　金：上海市教育委员会科研创新计划项目(2017-01-07-00-E00064)。

摘　　要：目的观察骨痛灵方对肺癌骨转移疼痛模型小鼠胫骨的核因子κB受体活化因子配体(RANKL)、骨保护素(OPG)和单核细胞趋化蛋白-1(MCP-1)、环氧化酶2(COX-2)、肿瘤坏死因子α(TNF-α)表达的影响,探讨骨痛灵方治疗肺癌骨转移疼痛的潜在作用机制。方法80只小鼠随机分为假手术组、模型组、中药组、西药组和中西药组,每组16只。建立Lewis肺癌骨转移小鼠模型,中药组以骨痛灵方煎剂灌胃,西药组予以唑来膦酸腹腔注射。中西药组则给予骨痛灵方水煎液和唑来膦酸水溶液,药物用法用量与中药组、西药组相同。假手术组和模型组均给予等剂量的0.9%氯化钠注射液。采用RT-PCR法和Western Blot检测小鼠胫骨RANKL、OPG mRNA和蛋白表达,用酶联免疫吸附法(ELISA法)检测小鼠胫骨MCP-1、COX-2和TNF-α表达情况。结果与假手术组相比模型组RANKL mRNA和蛋白表达增加,OPG mRNA和蛋白表达减少,差异均具有统计学意义(P<0.05)。与模型组相比,中药组、西药组和中西药组均能显著降低RANKL mRNA和蛋白表达,升高OPG mRNA和蛋白表达,降低RANKL/OPG比值,差异均有统计学意义(P<0.05)。与假手术组相比,模型组COX-2、TNF-α、MCP-1表达增加,差异具有统计学意义(P<0.05)。与模型组比较,中药组、西药组和中西药组的COX-2、TNF-α、MCP-1表达均明显降低,差异具有统计学意义(P<0.05),且中西药组对于降低COX-2、TNF-α、MCP-1的表达具有协同作用(P<0.05)。结论骨痛灵方可能通过抑制肺癌骨转移的炎性反应,抑制破骨细胞的活化,发挥止痛作用。Objective To observe the nuclear factorκB receptor activating factor ligand(RANKL),osteoprotegerin(OPG),and monocyte chemoattractant protein-1(MCP-1)of Gutongling recipe on the tibia of lung cancer bone metastasis pain model mice,The influence of cyclooxygenase 2(COX-2)and tumor necrosis factor alpha(TNF-α)expression,to explore the potential mechanism of action of Gutonglingfang in the treatment the pain of lung cancer bone metastasis.Methods The bone metastasis mouse model of lung cancer was built with the Lewis cells suspension,which was injected into the left tibia medullary cavity with microsyringe.80 C57BL/6J mice were randomly divided into five groups:sham-operated group(SG),model group(MG),Chinese medicine group(CG),west medicine group(WG),Chinese and west medicine combination group(CWG),each goup of 16 mice.The mice of CG group were intragastred with Gu-tong-ling prescription,while the mice of WG group were injected intraperitoneal with zoledronic acid.Both the SG and the MG group were given the same amount of 0.9%sodium chloride injection.In CWG,the mice were received the two medicine with the treatment course of three weeks.RT-PCR and Western blot were used to detect the expression of RANKL,OPG mRNA and protein in mouse tibia,and the expression of MCP-1,COX-2 and TNF-αin mouse tibia was detected by enzyme-linked immunosorbent assay(ELISA).Results Compared with SG,the mRNA and protein expressing of RANKL in MG are higher,the mRNA and protein expressing of OPG in MG are lower,and the differences have statisticalsignificance(both P<0.05).Compared with the MG,the CG,WG and CWG both have lower expressing of RANKL mRNA and protein,and higher expressing of OPG mRNA and protein,lower RANKL/OPG ratio,and the differences have statistical significance(both P<0.05).Compared with the SG,the expressing of COX-2,TNF-αand MCP-1 in the MG are both much higher than that of SG,and the differences have statistical significance(both P<0.05).Compared with MG,the expressing of COX-2,TNF-αand MCP-1 in the CG,WG and CWG ar

关 键 词：骨癌痛 肺癌骨转移 骨痛灵方 炎性因子 核因子ΚB受体活化因子配体 

分 类 号：R285.5[医药卫生—中药学]