摘要
目的从分子生物学的角度,探讨降钙素基因相关肽(calcitonin gene-related peptide,CGRP)在慢性胃溃疡模型小鼠穴位敏化过程中的作用。方法将C57BL/6小鼠随机分为对照组和模型组,每组15只,将冰乙酸溶液于胃小弯近幽门处注射到胃壁肌层近黏膜下层构建慢性胃溃疡模型,随后在小鼠尾静脉上注射伊文思蓝,观察穴位敏化点的体表分布规律;HE染色法观察胃部组织在造模前后的病理形态变化;通过体表痛敏行为学实验,观察小鼠痛觉阈值的改变;蛋白质印迹法、免疫荧光染色法检测穴位敏化后脊髓、背根神经节(dorsal root ganglion,DRG)和皮肤中的CGRP表达。结果通过向小鼠胃部注射冰乙酸可诱导其形成符合胃溃疡典型病理特征的动物模型;对照组EB渗出点较少,模型组EB渗出点分散于脊髓T_(1)~T_(13)节段支配区域,其中以T_(9)~_(T11)区域最为集中,且其伊文思蓝渗出点压痛阈较造模前显著降低(P<0.01)。穴位敏化后模型组的脊髓、DRG和皮肤中的CGRP蛋白和荧光蛋白阳性表达显著高于对照组(P<0.01)。结论冰乙酸诱导小鼠形成胃溃疡后,其胃部组织病变所对应的体表敏化区、相应的脊髓T9~T11节段和DRG神经元中CGRP表达增高,可能是引起外周和中枢痛觉敏化,使体表局部出现神经源性炎性反应,介导穴位敏化的原因。
Objective To discuss the role of calcitonin gene-related peptide(CGRP)in acupoint sensitization in chronic gastric ulcer mice from the perspective of molecular biology.Method C57BL/6 mice were randomly divided into a control group and a model group,with 15 mice in each group.The chronic gastric ulcer model was established by injection of glacial acetic acid into the submucosal layer of the gastric wall near the pylorus in the lesser curvature,followed by Evans blue injection into the tail vein of mice to observe the distribution pattern of acupoint sensitization spots on the body surface.Pathological changes in gastric tissues after modeling were observed by hematoxylin-eosin staining.Changes in the pain threshold of mice were observed by body surface hyperalgesia-like behavior tests.The expression of CGRP in the spinal cord,dorsal root ganglion(DRG),and skin after acupoint sensitization was detected by Western blotting and immunofluorescence staining.Result Injecting glacial acetic acid into the stomach of mice could induce a model conforming to the typical pathological features of gastric ulcer.The control group had fewer EB exudate spots,while the model group had EB exudate spots scattered in the dominant regions of T_(1)-T_(13) segments of the spinal cord,predominantly concentrated in the T_(9)-T_(11) regions,and a significantly lower pressing pain threshold was found at the Evans blue exudate spots compared with that before modeling (P<0.01). After acupoint sensitization, the CGRP protein levels and positive expression of fluorescent protein in the spinal cord, DRG, and skin were significantly higher in the model group than in the control group (P<0.01). Conclusion After the establishment of gastric ulcers induced by glacial acetic acid in mice, the expression levels of CGRP in the sensitized regions on the body surface, the corresponding T9-T11 spinal cord segments, and DRG neurons corresponding to the gastric lesions were significantly accelerated, which may be responsible for peripheral and central hyperalgesia, topical neurogenic inflammatory responses on the body surface, and mediating acupoint sensitization.
作者
杨雪
郑璇
袁伟
吴宇蔚
刁志君
薛思梦
乔海法
YANG Xue;ZHENG Xuan;YUAN Wei;WU Yuwei;DIAO Zhijun;XUE Simeng;QIAO Haifa(Shaanxi University of Chinese Medicine,Xianyang 712046,China;Shaanxi Key Laboratory of Acupuncture and Medicine,Xianyang 712046,China;Shaanxi Administration of Traditional Chinese Medicine,Xianyang 712046,China)
出处
《上海针灸杂志》
CSCD
2024年第1期81-88,共8页
Shanghai Journal of Acupuncture and Moxibustion
基金
国家自然科学基金项目(81873385,82074557)
陕西省教育厅项目(18JK0213)
陕西省科技厅项目(2019JM-399)。
关键词
胃溃疡
穴位敏化
痛阈
降钙素基因相关肽
小鼠
Stomach ulcer
Acupoint sensitization
Pain threshold
Calcitonin gene-related peptide
mice
