基于对背根神经节中神经生长因子的调控探究华蟾素治疗骨癌痛的机制  

Mechanism of cinobufagin in treatment of bone cancer pain based on regulation of nerve growth factor in dorsal root ganglion

作　　者：刘亚丽 刘丹 张文 陈涛[1,4] 胡卫[1,4] LIU Ya-li;LIU Dan;ZHANG Wen;CHEN Tao;HU Wei(Medical College,China Three Gorges University,Yichang 443003,China;Huanggang Traditional Chinese Medicine Hospital,Huanggang 438000,China;Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China;Third-grade Pharmacological Laboratory on Traditional Chinese Medicine,State Administration of Traditional Chinese Medicine,China Three Gorges University,Yichang 443003,China)

机构地区：[1]三峡大学医学院,湖北宜昌443003 [2]黄冈市中医医院,湖北黄冈438000 [3]上海中医药大学附属龙华医院,上海200032 [4]三峡大学国家中医药管理局中药药理科研三级实验室,湖北宜昌443003

出　　处：《中草药》2022年第17期5427-5432,共6页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(81503381)。

摘　　要：目的 基于对骨癌痛大鼠背根神经节(dorsal root ganglion,DRG)中神经生长因子(nerve growth factor,NGF)的调控,探讨中药华蟾素治疗骨癌痛的机制。方法 筛选痛阈值满足条件的SD雌性大鼠构建骨癌痛模型,随机分为正常组、假手术组、模型组、华蟾素组、华蟾素+NGF组和anti-NGF组。华蟾素组、华蟾素+NGF组在造模第7天开始ip华蟾素,其余各组ip等体积的生理盐水;华蟾素+NGF组、anti-NGF组在造模第12天鞘内插管给药,造模21 d后处理大鼠。检测大鼠造模前后痛阈值变化;采用Western blotting法检测各组大鼠DRG中原肌球蛋白受体激酶A(tropomyosin receptor kinase A,TrkA)、p75神经营养受体(p75 neurotrophy receptor,p75NTR)、μ阿片受体(μopioid receptor,MOR)和神经递质P物质(substance P,SP)蛋白表达;采用免疫组化法检测各组大鼠DRG中MOR的表达;采用免疫荧光双标检测各组大鼠DRG中MOR与TrkA共表达情况。结果 与模型组比较,华蟾素明显提高大鼠机械痛阈值与热痛阈值(P<0.01);华蟾素组抑制大鼠DRG中TrkA、p75NTR和SP蛋白表达(P<0.05、0.01),上调MOR蛋白表达(P<0.01)。华蟾素+NGF组激活大鼠DRG中TrkA、p75NTR和SP的表达(P<0.01),抑制MOR的表达(P<0.05)。结论 华蟾素可能通过调节内源性NGF抑制TrkA、p75NTR的表达,激活MOR,并能够减少SP的释放,最终参与骨癌痛的镇痛过程。Objective To investigate the mechanism of cinobufagin in the treatment of bone cancer pain based on regulation of nerve growth factor(NGF) in dorsal root ganglion(DRG) of rats with bone cancer pain. Methods SD female rats with pain thresholds meeting the conditions were screened to construct a bone cancer pain model and randomly divided into normal group, sham group,model group, cinobufagin group, cinobufagin + NGF group and anti-NGF group. Cinobufagin group and cinobufagin + NGF group were ip cinobufagin on 7th day after modeling, and the other groups were given an equal volume of normal saline;Cinobufagin + NGF group and anti-NGF group were administered by intrathecal intubation on 12th day after modeling. After intubation administration,rats were treated for 21 d after modeling. The changes of pain threshold before and after the rat model were detected;Western blotting was used to detect tropomyosin receptor kinase A(TrkA) and p75 neurotrophy receptor(p75NTR), μ opioid receptor(MOR) and neurotransmitter substance P(SP) protein expressions in DRG of rats in each group;Immunohistochemistry was used to detect the expression of MOR in DRG of rats in each group;Immunofluorescence double-labeling was used to detect the co-expression of MOR and TrkA in DRG of rats in each group. Results Compared with model group, cinobufacin significantly increased the mechanical pain threshold and thermal pain threshold(P < 0.01);Cinobufacin inhibited TrkA, p75NTR and SP protein expressions in DRG of rats(P < 0.05, 0.01), up-regulated MOR protein expression(P < 0.01). Cinobufacin + NGF activated the protein expressions of TrkA,p75NTR and SP in DRG of rats(P < 0.01), and inhibited the expression of MOR(P < 0.05). Conclusion Cinobufacin may activate MOR by inhibiting the expression of NGF and its receptors TrkA and p75NTR, and thus reduce the release of SP, and finally participate in analgesic process of bone cancer pain.

关 键 词：华蟾素 骨癌痛 神经生长因子 Μ阿片受体 原肌球蛋白受体激酶A p75神经营养受体 

分 类 号：R285.5[医药卫生—中药学]