普鲁卡因调节HMGB1-RAGE信号通路对脑缺血再灌注大鼠的神经保护作用研究

Neuroprotective Effect of Procaine on Cerebral Ischemia-reperfusion Rats by Regulating HMGB1-RAGE Signaling Pathway

目的探讨普鲁卡因(PCA)调节高迁移率族蛋白1-晚期糖基化终末产物受体(HMGB1-RAGE)信号通路对脑缺血/再灌注(CI/R)大鼠的神经保护作用。方法采用线栓法制备CI/R大鼠模型,缺血处理2 h,再灌注24 h。将大鼠随机分为假手术组,模型组(CI/R组),普鲁卡因低、中、高剂量组(PCA-L组、PCA-M组、PCA-H组),普鲁卡因高剂量+HMGB1重组蛋白组(PCA-H+HMGB1组),各组n=10。对大鼠进行神经功能评分,苏木精-伊红染色法观察大鼠脑组织海马CA1区组织病理变化,Tunel染色法检测大鼠脑组织海马CA1区神经元凋亡,酶联免疫吸附(ELISA)法检测大鼠脑组织中TNF-α、IL-1β水平;免疫组化法检测大鼠脑组织NGF表达;Western blot法检测大鼠脑组织中HMGB1、RAGE表达。结果与假手术组比较,CI/R组脑组织损伤严重,神经功能评分、神经元凋亡率、IL-1β及TNF-α水平、NGF表达、HMGB1及RAGE表达均显著升高(均P<0.05);与CI/R组比较,PCA-L组、PCA-M组、PCA-H组大鼠脑组织损伤相对减轻,神经功能评分、神经元凋亡率、IL-1β及TNF-α水平、HMGB1及RAGE表达均呈显著降低,NGF表达显著升高,呈剂量依赖效应(P<0.05);与PCA-H组比较,PCA-H+HMGB1组大鼠脑组织损伤加重,神经功能评分、神经元凋亡率、IL-1β及TNF-α水平、HMGB1及RAGE表达均显著升高,NGF表达显著降低(均P<0.05)。结论普鲁卡因对CI/R大鼠的神经具有一定的保护作用,其机制可能与抑制HMGB1-RAGE信号通路激活相关。

Aim To investigate the neuroprotective effect of procaine(PCA)on cerebral ischemiareperfusion(CI/R)rats by regulating the high-mobility group protein-advanced glycation end product receptor(HMGB1-RAGE)signaling pathway.Methods A rat model of cerebral ischemia-reperfusion was prepared using thread occlusion method,with ischemia treatment for 2 hours and reperfusion for 6 hours.The rats were randomly divided into a sham surgery group(sham group),a model group(CI/R group),procaine low,medium and high dose groups(PCA-L group,PCA-M group,PCA-H group),a high dose of procaine+HMGB1 recombinant protein group(PCA-HMGB1 group),10 rats in each group.Neurological function scoring was performed on these rats.HE staining method was applied to observe the pathological changes in the hippocampal CA1 region of rat brain tissue.Tunel staining method was applied to detect neuronal apoptosis in the hippocampal CA1 region of rat brain tissue.Enzyme linked immunosorbent assay(ELISA)was applied to detect levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in rat brain tissue.Immunohistochemical method was applied to detect the expression of nerve growth factor(NGF)in rat brain tissue,and Western blot method was applied to detect the expression of HMGB1 and RAGE in rat brain tissue.Results Compared with the sham group,the brain tissue damage of rats in the CI/R group was severe,and the neurological function score,neuronal apoptosis rate,levels of IL-1βand TNF-α,the expression of NGF,HMGB1 and RAGE were obviously increased(P<0.05).Compared with the CI/R group,the brain tissue damage of rats in the PCA-L,PCA-M,and PCA-H groups was relatively reduced,the neurological function score,neuronal apoptosis rate,levels of IL-1βand TNF-α,the expression of HMGB1 and RAGE were obviously reduced,and the expression of NGF was obviously increased,in a dose-dependent manner(P<0.05).Compared with the PCA-H group,the brain tissue damage in the PCA-HMGB1 group was aggravated,the neurological function score,neuronal apoptosis rate,levels of IL-1βand TNF-α,the expression of HMGB1 and RAGE were obviously increased,and the expression of NGF was obviously reduced(P<0.05).Conclusion Procaine has a protective effect on the nerves of cerebral ischemia-reperfusion rats,and its mechanism may be related to inhibiting the activation of HMGB1-RAGE signaling pathway.