天麻素对新生大鼠缺氧缺血性脑损伤后星形胶质细胞表型和晚期糖基化终末产物受体表达的影响  

Effects of gastrodin on astrocyte phenotype and the receptor of advanced glycation endproducts expression after hypoxic⁃ischemic brain damage in neonatal rats

作　　者：王鹏翔 任雪琪 左涵珺 万程[2] 石金沙 石浩龙 赵敏[1] 李娟娟[1] WANG Peng-xiang;REN Xue-qi;ZUO Han-jun;WAN Cheng;SHI Jin-sha;SHI Hao-long;ZHAO Min;LI Juan-juan(Department of Human Anatomy,Histology and embryology,Kunming Medical University,Kunming 650500,China;Department of Medical Imaging,the First Affiliated Hospital of Kunming Medical University,Kunming 650032,China)

机构地区：[1]昆明医科大学人体解剖学与组织胚胎学系,昆明650500 [2]昆明医科大学第一附属医院医学影像科,昆明650032

出　　处：《解剖学报》2024年第6期677-684,共8页Acta Anatomica Sinica

基　　金：国家自然科学基金(31960194);云南省科技厅-昆明医科大学应用基础研究联合专项重点项目[2019FE001(-003)];云南省神经系统疾病临床医学中心子课题(ZX2019030501)。

摘　　要：目的研究新生大鼠缺氧缺血性脑损伤(HIBD)后反应性星形胶质细胞表型和晚期糖基化终末产物受体(RAGE)的表达及天麻素(GAS)干预对其的影响。方法利用48只新生3 d的SD大鼠构建HIBD模型,随机分为假手术组(sham)、HIBD模型组(HIBD)及HIBD后GAS(100 mg/kg)干预组(HIBD+GAS)。Western blotting和免疫组织化学染色检测HIBD后1 d、3 d组缺血侧大脑胼胝体区A1型星形胶质细胞标志物C3、A2型星形胶质细胞标志物S100A10、RAGE、肿瘤坏死因子α(TNF-α)、脑源性神经营养因子(BDNF)和胰岛素样生长因子1(IGF-1)的表达。体外复制TNC-1细胞氧糖剥夺模型(OGD),随机分为对照组(Ctrl)、氧糖剥夺组(OGD)、氧糖剥夺后GAS干预(0.34 mmol/L)组(OGD+GAS)和单纯GAS组(GAS)。Western blotting和免疫荧光染色检测各实验组RAGE、TNF-α、BDNF和IGF-1的蛋白表达。结果与假手术组相比,HIBD后1、3 d缺血侧胼胝体区C3、S100A10、RAGE、TNF-α和IGF-1蛋白表达明显升高(P<0.05);1 d组BDNF蛋白表达降低,而3 d组表达升高(P<0.05)。GAS干预后1 d、3 d组C3、RAGE和TNF-α表达较HIBD组降低(P<0.05),而BDNF和IGF-1蛋白表达进一步升高(P<0.05);GAS干预后3 d组S100A10的表达较HIBD组升高(P<0.05)。且HIBD后1 d、3 d组C3、S100A10、RAGE的免疫组织化学检测结果与Western blotting结果一致。此外,在OGD激活的星形胶质细胞中RAGE、TNF-α表达显著升高(P<0.05),GAS干预后两者表达显著降低(P<0.05),而BDNF、IGF-1表达显著升高(P<0.05)。结论GAS可抑制HIBD后星形胶质细胞中RAGE信号的上调,抑制A1型星形胶质细胞和炎性因子的表达,促进A2型星形胶质细胞和营养因子的表达,发挥神经保护功能。Objective To investigate the activated phenotype and the expression of the receptor of advanced glycation endproducts(RAGE)of astrocytes after hypoxic⁃ischemic brain damage(HIBD)in neonatal rats and the effects of gastrodin(GAS)intervention on them.Methods Totally 48 neonatal 3 days SD rats were used to construct HIBD model and randomly divided into sham group,HIBD group and HIBD+GAS group(100 mg/kg),and the expressions of A1 type astrocyte marker C3,A2 type astrocyte marker S100A10,RAGE,tumor necrosis factor⁃α(TNF⁃α),brain⁃derived neurotrophic factor(BDNF),and insulin⁃like growth factor(IGF⁃1)in the corpus callosum of the ischemic side were detected by Western blotting and immunohistochemical staining on day 1 and day 3 after HIBD.TNC⁃1 cells were divided into control group,oxygen glucose deprivation(OGD)group,OGD+GAS(0.34 mmol/L)group and GAS group,and then the protein expressions of RAGE,TNF⁃α,BDNF and IGF⁃1 were detected by Western blotting and immunofluorescence.Results In vivo,Western blotting showed that compared with the sham group,the protein expression levels of C3,S100A10,RAGE,TNF⁃αand IGF⁃1 in the 1 day and 3 days groups after HIBD group in 1 day group were significantly higher than those in the sham group(P<0.05),but the protein expression level of BDNF decreased in 1 day group and increased in 3 days group(P<0.05).Compared with the HIBD group,the C3,RAGE and TNF⁃αprotein expression levels were significantly attenuated in the HIBD+GAS group(P<0.05),and the protein expression levels of BDNF and IGF⁃1 further increased(P<0.05).The protein expression of S100A10 in the 3 days group was higher than that in the HIBD group after GAS treatment(P<0.05).The immunohistochemical staining results of C3,S100A10,and RAGE in the 1 day and 3 days groups after HIBD were consistent with Western blotting results.Furthermore,the protein expressions of RAGE and TNF⁃αwere significantly enhanced in OGD⁃stimulated astrocytes(P<0.05).After GAS intervention,while the expressions of both RAGE

关 键 词：晚期糖基化终末产物受体 星形胶质细胞 缺氧缺血性脑损伤 天麻素 免疫印迹法 新生大鼠 

分 类 号：R329.1[医药卫生—人体解剖和组织胚胎学]