基于对骨癌痛大鼠脊髓TAK1/JNK/c-Jun信号通路的调控探讨华蟾素镇痛机制  

Investigation on the Analgesic Effect of Cinobufagin based on Regulation of the TAK1/JNK/c-Jun Signaling Pathway in the Spinal Cord of Rats with Bone Cancer Pain

作　　者：刘丹 焦良波[2] 陈涛[3] 付烊 胡卫[3] LIU Dan;JIAO Liang-bo;CHEN Tao;FU Yang;HU Wei(Huanggang Traditional Chinese Medicine Hospital,Huanggang 438000,China;The Second People's Hospital Affiliated to Three Gorges University,Yichang Second People's Hospital,Yichang 443002,China;Third-Grade Pharmacological Laboratory on Traditional Chinese Medicine,State Administration of Traditional Chinese Medicine,Medical College of Three Gorges University,Yichang 443000,China;Xiangyang Hospital of Traditional Chinese Medicine,Xiangyang 441000,China)

机构地区：[1]黄冈市中医医院,湖北黄冈438000 [2]三峡大学附属第二人民医院(宜昌市第二人民医院),湖北宜昌443002 [3]三峡大学医学院,国家中医药管理局中药药理(肿瘤)科研三级实验室,湖北宜昌443000 [4]襄阳市中医医院,湖北襄阳441000

出　　处：《中国药学杂志》2021年第2期108-114,共7页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金项目资助(81503381)。

摘　　要：目的基于对骨癌痛大鼠脊髓星形胶质细胞TAK1/JNK/c-Jun信号通路的调控,探讨华蟾素缓解骨癌痛的作用机制。方法选取雌性SD大鼠,随机分成6组,在造模前后检测行为学指标,最后一次行为学检测结束后处理大鼠,Western blot法检测相关蛋白的表达情况,免疫荧光双标检测脊髓c-jun氨基末端激酶(c-Jun N-terminal kinases,JNK)蛋白与胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)的共定位情况,ELISA法检测脊髓相关细胞因子的含量。结果与模型组相比,华蟾素组大鼠的痛阈值上升(P<0.01或P<0.05);鞘内插管组大鼠的痛阈值未见明显变化;抑制剂组进行鞘内给药后,大鼠的痛阈值明显升高(P<0.05)。Western blot结果显示,模型组大鼠脊髓中GFAP、p-TAK1、p-JNK、p-c-Jun蛋白的表达增多(P<0.01);华蟾素能显著降低模型组大鼠脊髓中GFAP、p-TAK1、p-JNK、p-c-Jun蛋白的表达(P<0.01或P<0.05);抑制剂组大鼠脊髓中p-JNK、p-c-Jun蛋白的表达降低(P<0.05),而GFAP、p-TAK1蛋白的表达未见明显差异。免疫荧光结果显示,脊髓中p-JNK与GFAP共表达。ELISA结果显示,模型组大鼠脊髓中肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、CCL2的表达较假手术组增多(P<0.01);华蟾素组及抑制剂组脊髓中TNF-α、IL-1β、CCL2的表达较模型组减少(P<0.01);抑制剂组CCL2的表达较华蟾素组降低(P<0.05),而TNF-α、IL-1β的表达未见明显差异。结论华蟾素可能是通过抑制脊髓星形胶质细胞中TAK1/JNK/c-Jun信号通路相关蛋白的活化,减少细胞因子释放,从而发挥缓解骨癌痛的作用。OBJECTIVE To investigate the analgesic effect of cinobufagin based on regulation of the TAK1/JNK/c-Jun signaling pathway in the spinal cord of rats with bone cancer pain.METHODS Seventy SD rats were randomly divided into six groups,and pain behavior was tested before and after modeling,respectively.The expressions of TAK1,JNK,c-jun,p-tak1,p-jnk,and p-c-jun in the spinal cord were detected by Western blot,while co-expression of JNK and GFAP in the spinal cord was detected by double-labelling immunofluorescence and the contents of IL-1β,TNF-α,and CCL2 were detected by ELISA.RESULTS The pain threshold was decreased in the model group,but increased in the cinobufagin group and the celecoxib group.After intrathecal administration of SP600125,the pain threshold was significantly higher than that of the saline group(P<0.05).While the pain threshold of each group(celecoxib group,inhibitor group)was significantly lower than that of the cinobufagin group(P<0.05).The protein levels of GFAP,p-TAK1,p-JNK and p-c-Jun in the spinal cord of the model group were higher than that of the sham group(P<0.01).After injected with cinobufagin,the protein levels were significantly decreased compared with the model group(P<0.05).The expression levels of GFAP and p-TAK1 in the inhibitor group had no change(P<0.05),while the expressions of p-JNK and p-c-Jun were decreased(P<0.05).ELISA results showed that the contents of IL-1β,TNF-α,and CCL2 were significantly increased in the model group compared with the sham group(P<0.01).The contents of IL-1β,TNF-α,and CCL2 of the cinobufagin group and inhibitor group were lower than that of the model group(P<0.01).Compared with the cinobufagin group,the expression of CCL2 in the inhibitor group was significantly decreased(P<0.05),while the contents of IL-1βand TNF-αhad no difference(P<0.05).CONCLUSION Cinobufagin can inhibit the expression of proteins related to the TAK1/JNK/c-Jun signaling pathway in the spinal cord in the rat model of bone cancer pain,ultimately decrease the contents of IL-1�

关 键 词：华蟾素 骨癌痛 TAK1/JNK/c-Jun信号通路 镇痛 

分 类 号：R965[医药卫生—药理学]