加巴喷丁减轻骨折模型大鼠焦虑情绪及海马神经炎症的作用机制研究

Mechanism of gabapentin alleviating anxiety and hippocampal neuroinflammation induced by fracture model in rats

目的探析加巴喷丁对骨折模型大鼠焦虑情绪、海马神经炎症、人核因子κB(NF-κB)/肿瘤坏死因子α(TNF-α)信号通路的影响。方法自2022年1―6月,选择健康雄性无特定病原体(SPF)SD大鼠48只,采用随机数字表法分为假手术组、模型组、治疗组。假手术组正常饲养,未做任何处理,其余两组大鼠均进行骨折造模。其中治疗组给予加巴喷丁治疗,模型组给予等量生理盐水干预,连续给药12 d。采用高架十字迷宫实验评估大鼠焦虑情绪;采用酶联免疫吸附(ELISA)检测白细胞介素-4(IL-4)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、超氧化物歧化酶(SOD)、丙二醛(MDA),采用比色法检测谷胱甘肽过氧化物酶(GSH-Px)。采用苏木素伊红(HE)染色评估海马组织病理形态特征;采用蛋白质印迹法检测TNF-α、NF-κB蛋白表达量。结果假手术组、治疗组、模型组总穿臂次数(TE)、进入比例(OE)、停留时间比例(OT)比较,假手术组大鼠TE[(12.31±2.59)次比(10.29±2.37)次、(7.25±2.16)次]、OE[(40.25±5.41)%比(37.02±5.13)%、(32.77±5.43)%]、OT[(23.67±4.22)%比(21.94±5.31)%、(16.47±2.35)%]均高于治疗组、模型组大鼠,治疗组大鼠TE、OE、OT均高于模型组(P<0.05)。假手术组大鼠IL-4[(41.23±5.37)ng/L比(50.23±6.78)ng/L、(59.44±6.92)ng/L]、IL-6[(101.45±12.36)ng/L比(114.32±12.74)ng/L、(135.49±16.78)ng/L]、IL-1β[(87.64±8.74)ng/L比(101.23±10.45)ng/L、(110.34±10.25)ng/L]均低于治疗组、模型组大鼠,治疗组大鼠IL-4、IL-6、IL-1β均低于模型组(P<0.05)。假手术组大鼠GSH-Px、SOD均高于治疗组、模型组大鼠,MDA低于两组;且治疗组大鼠GSH-Px、SOD均高于模型组,MDA低于该组(P<0.05)。假手术组大鼠NF-κB、TNF-α蛋白表达均低于治疗组、模型组大鼠,治疗组大鼠NF-κB、TNF-α蛋白表达均低于模型组(P<0.05)。结论骨折导致大鼠焦虑情绪增加,而加巴喷丁可以有效削弱骨折大鼠的焦虑情绪,可能与加巴喷丁减弱海马炎症反应有关,从而增强NF-κB/TNF-α信号通路对海马神经的保护作用。

Objective To investigate the effect of gabapentin on anxiety,hippocampal neuroinflammation and human nuclear factor kappa B(NF-κB)/tumor necrosis factorα(TNF-α)signaling pathway induced by fracture model in rats.Methods From January to June 2022,forty-eight healthy male specific pathogen-free(SPF)SD rats were selected and assigned into sham operation group,model group,and treatment group by random number table method.The rats in the sham operation group were fed normally without any treat⁃ment,while the other two groups of rats were subjected to fracture modeling.The treatment group was given gabapentin,and the model group was given the same amount of normal saline for continuous administration for 12 days.The elevated plus maze test was used to evaluate rat anxiety,enzyme linked immunosorbent assay(ELISA)to detect interleukin-4(IL-4),interleukin-6(IL-6),interleukin-1β(IL-1β),superoxide dismutase(SOD),malondialdehyde(MDA),and colorimetry to detect glutathione peroxidase(GSH-Px).Hematoxy⁃lin eosin(HE)staining was used to evaluate the histopathological characteristics of hippocampus,and Western blotting was used to de⁃tect the expression levels of TNF-αand NF-κB.Results In the sham operation group,total pass times(TE)[(12.31±2.59)vs.(10.29±2.37),(7.25±2.16)],entry ratio(OE)[(40.25±5.41)%vs.(37.02±5.13)%,(32.77±5.43)%],and residence time ratio(OT)[(23.67±4.22)%vs.(21.94±5.31)%,(16.47±2.35)%]were higher than those in the treatment group and model group.The TE,OE and OT of rats in the treatment group were higher than those in the model group(P<0.05).The IL-4[(41.23±5.37)ng/L vs.(50.23±6.78)ng/L,(59.44±6.92)ng/L],IL-6[(101.45±12.36)ng/L vs.(114.32±12.74)ng/L,(135.49±16.78)ng/L],and IL-1β[(87.64±8.74)ng/L vs.(101.23±10.45)ng/L,(110.34±10.25)ng/L]in the sham operation group were lower than those in the treatment group and the model group,and the IL-4,IL-6 and IL-1βin the treatment group were lower than those in the model group(P<0.05).The GSH-Px and SOD of the rats in the sham operation group were higher than those of the treatment group and the model group,while the MDA in the sham operation group was lower than that of the other two groups,and the GSH-Px and SOD of the rats in the treatment group were lower than those of the model group,and the MDA was lower than that of the model group(P<0.05).The protein expressions of NF-κB and TNF-αin the sham operation group were lower than those in the treatment group and the model group,and the protein expressions of NF-κB and TNF-αin the treatment group were lower than those in the model group(P<0.05).Conclusion Fractures lead to increased anxiety in rats,while gabapentin can effectively attenuate anxiety in fractured rats,which may be related to gabapentin attenuating hippocam⁃pal inflammatory response,thereby enhancing the protective effect of NF-κB/TNF-αsignaling pathway on hippocampal nerves.