[1]罗琴琴,鲁叶云.骨痛灵方对骨癌痛小鼠ERK5信号通路的作用机制研究[J].西部中医药,2025,38(10):7-12.[doi:10.12174/j.issn.2096-9600.2025.10.02]
　LUO Qinqin,LU Yeyun.Mechanism Study on Gutongling Prescription on ERK5 Signaling Pathway in Bone Cancer Pain Mice[J].Western Journal of Traditional Chinese Medicine,2025,38(10):7-12.[doi:10.12174/j.issn.2096-9600.2025.10.02]
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骨痛灵方对骨癌痛小鼠ERK5信号通路的作用机制研究

《西部中医药》[ISSN:2096-9600/CN:62-1204/R]

卷:
38
期数:
2025年10期
页码:
7-12
栏目:
基础研究
出版日期:
2025-10-15

文章信息/Info

Title:
Mechanism Study on Gutongling Prescription on ERK5 Signaling Pathway in Bone Cancer Pain Mice
作者:
罗琴琴1, 鲁叶云2
1.武汉市中西医结合医院，湖北 武汉 430000
2.上海市第十人民医院崇明分院，上海 202150
Author(s):
LUO Qinqin1, LU Yeyun2
1.Wuhan Hospital of Traditional Chinese and Western Medicine, Wuhan 430000, China
2.Chongming Branch of Shanghai Tenth People's Hospital, Shanghai 202150, China
关键词:
骨癌痛; 破骨细胞; ERK5; 骨痛灵方; 动物试验
Keywords:
bone cancer pain; osteoclast; ERK5; prescription; animal experiment
分类号:
R273
DOI:
10.12174/j.issn.2096-9600.2025.10.02
文献标志码:
A
摘要:
目的观察骨痛灵方对肺癌骨转移疼痛小鼠细胞外信号调节激酶5（extracellular signal regulated kinase，ERK5）通路的影响。 方法将40只C57BL/6J小鼠按随机数字表法分为假手术组、模型组、中药组和抑制剂组，每组10只。模型组、中药组和抑制剂组小鼠以胫骨骨髓腔内注射Lewis肺癌细胞建立骨转移癌痛模型，假手术组小鼠注射等量PBS溶液。中药组以骨痛灵方水煎剂溶液灌胃，每日1次；抑制剂组皮下注射ERK5抑制剂（XMD8-92），每日2次；假手术组皮下注射生理盐水，每日2次；模型组干预方法同假手术组。各组小鼠均连续干预21天。造模前及造模后第7、14、21天通过Von Frey纤维丝测定小鼠机械性痛觉超敏机械性痛阈值（paw withdrawal mechanical threshold，PWMT）；造模前及造模后第6、13、20天采用全自动热痛仪检测小鼠热痛觉超敏（paw withdrawal thermal latency，PWTL）。干预21天后，对假手术组和模型组小鼠进行X线检测、HE染色以及抗酒石酸酸性磷酸酶（tartrate-resistant acid phosphatase，TRAP）染色；并采用蛋白免疫印迹法检测各组小鼠胫骨组织ERK5、p-ERK5蛋白表达情况，qRT-PCR法检测各组小鼠胫骨组织组织蛋白酶K（cathepsin K，Ctsk）和基质金属蛋白酶9（matrix metalloprotein 9，MMP9）mRNA表达情况。 结果造模后第7天（6天）、第14天（13天）、第21天（20天），模型组小鼠PWMT和PWTL均低于假手术组（P＜0.05）。造模后第7天，模型组、中药组、抑制剂组PWMT和PWTL比较差异均无统计学意义（P＞0.05）。造模后第14、21天，与模型组比较，中药组和抑制剂组PWMT和PWTL均升高（P＜0.05）；与中药组比较，抑制剂组PWMT和PWTL均升高（P＜0.05）。胫骨X线结果显示，假手术组小鼠胫骨平台完整，骨皮质光滑连续，骨髓腔密度均匀；模型组可见溶骨性改变。HE染色结果显示，假手术组小鼠胫骨平台完整，骨皮质连续，骨小梁丰富，骨髓腔内充满骨髓细胞；模型组胫骨平台均出现严重破损，骨皮质变薄且不连续，骨小梁明显减少，骨髓腔内正常骨髓细胞少见。各组小鼠胫骨组织ERK5蛋白表达无明显差异（P＞0.05）。模型组小鼠胫骨p-ERK5蛋白表达与假手术组相比明显增加，p-ERK5/ERK5比值增高。与模型组相比较，中药组和抑制剂组p-ERK5蛋白表达及p-ERK5/ERK5比值均降低（P＜0.05）；抑制剂组p-ERK5蛋白表达以及p-ERK5/ERK5比值与中药组相比较明显降低（P＜0.05）。与假手术组比较，模型组TRAP阳性细胞数明显增多（P＜0.05）；与模型组比较，中药组和抑制剂组TRAP阳性细胞数量明显减少；与中药组比较，抑制剂组TRAP阳性细胞减少更明显（P＜0.05）。与假手术组比较，模型组小鼠胫骨Ctsk和MMP9 mRNA表达明显增加（P＜0.05）；与模型组比较，中药组和抑制剂组Ctsk和MMP9 mRNA表达均降低（P＜0.05）；与中药组比较，抑制剂组Ctsk和MMP9 mRNA表达明显降低（P＜0.05）。 结论骨痛灵方可有效缓解骨癌痛模型小鼠疼痛，其作用机制可能与骨痛灵方下调骨组织中破骨细胞的ERK5信号通路，抑制破骨细胞增殖活化有关。
Abstract:
ObjectiveTo observe the effects of Gutongling prescription on ERK5 pathway in mice suffering lung cancer and bone metastatic pain. MethodsForty C57BL/6J mice were assigned to sham operation group, the model group, herbal group and the inhibitor group according to random number table method with ten in each. The model group, herbal group and the inhibitor group were established into the models with bone metastatic pain by injecting Lewis lung cancer cells into the medullary cavity of the tibia, equivalent dose of PBS solution was administered to the mice in sham operation group. Herbal group was drenched with aqueous solution of the prescription, once each day; the inhibitor group received the injection of ERK5 inhibitor (XMD8-92), twice each day; subcutaneous injection of physiological saline was given to sham operation group, twice each day; the intervention methods of the model group was the same as sham operation group. The mice in different groups were intervened for 21 days consecutively. PWMT of the mice was detected via Von Frey fiber before modeling and on the 7th, 14th and 21st day after modeling; PWTL of the mice was measured using automatic thermal pain meter before modeling and on the 6th, 13th and 20th day after modeling. After 21 days of intervention, X-ray dectection, HE staining and TRAP staining were performed on sham operation group and the model group; Western blotting was used to detect the expression of ERK5, p-ERK5 protein in tibial bone tissue of the mice in different groups, qRT-PCR method was performed to the levels of Ctsk and MMP9 in tibial bone tissue of the mice in different groups. ResultsOn the 7th (6 days), 14th (13 days) and 21st (20 days) day after modeling, PWMT and PWTL of the model group were lower than these of sham operation group (P<0.05). PWMT and PWTL were compared between the model group, herbal group and inhibitor groups on the 7th day after modeling, and the differences were not statistically significant (P>0.05). On 14th and 21st day after modeling, the increase in PWMT and PWTL was found in herbal group and the inhibitor group when they were compared with the model group (P<0.05); compared with herbal group, PWMT and PWTL were elevated in the inhibitor group (P<0.05). Tibial X-ray results demonstrated that tibial plateau integrity, smooth and continuous cortical bone and uniform density of bone marrow cavity were present in sham operation group, while osteolytic changes were seen in the model group. HE staining results revealed that complete tibial plateau, smooth and continuous cortical bone, abundant bone trabeculae, bone marrow cells in the medullary cavity were found in sham operation group; while severe damage appeared in tibial plateau in the model group, thin and discontinuous cortical bone, bone trabeculae was reduced evidently, and normal bone marrow cells were infrequently observed in bone marrow cavity. The difference in ERK5 protein was insignficant between different groups (P>0.05). The expressions of p-ERK5 protein were evidently elevated in tibia of the model group when compared with sham operation group, p-ERK5/ERK5 ratio were lifted. The expressions of p-ERK5 protein and p-ERK5/ERK5 ratio were lowered when herbal group and the inhibitor group were compared with the model group (P<0.05); the reduction in the expressions of p-ERK5 protein and p-ERK5/ERK5 ratio was found when the inhibitor group was compared with herbal group (P<0.05). TRAP positive cells were increased when the model group was compared with sham operation group (P<0.05); compared with the model group, the numbers of TRAP positive cells were reduced obviously in herbal group and the inhibitor group; the decrease in TRAP positive cells was more significant when the inhibitor group was compared with herbal group (P<0.05). Upregulated expressions of Ctsk and MMP9 mRNA were observed in the model group when compared with sham operation group (P<0.05); compared with the model group, downregulated expressions of Ctsk and MMP9 mRNA were found in herbal group and the inhibitor group when compared with the model group (P<0.05); the expressions of Ctsk and MMP9 mRNA were evidently reduced in the inhibitor group when compared with herbal group (P<0.05). ConclusionGutongling prescription could effectively relieve pain in bone cancer pain mice models, and its mechanism might be related to the downregulation of ERK5 signaling pathway in osteoclast of bone tissue and the inhibition of the proliferation and activation of osteoclast by the prescription.

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备注/Memo

备注/Memo:
罗琴琴（1987—），女，博士学位，主治医师。研究方向：恶性肿瘤的中医诊治。国家自然科学基金青年科学基金项目（82305346）；湖北省自然科学基金（2020CFB359）。
更新日期/Last Update: 2025-10-15