栀子苷介导GLP-1R/Akt信号通路改善大鼠脑缺血再灌注损伤和神经元凋亡的研究  

Geniposide-mediated GLP-1R/Akt signaling pathway improves cerebral ischemia-reperfusion injury and neuronal apoptosis in rats

作　　者：谭慧敏 周亮[2] 邹慧禅 TAN Huimin;ZHOU Liang;ZOU Huichan(Department of Neurology,City Center Hospital of Zengcheng District in Guangzhou(Zengcheng Branch of Nanfang Hospital),Guangzhou 511300,China;Department of Neurology,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China;Department of Pharmacy,City Center Hospital of Zengcheng District in Guangzhou(Zengcheng Branch of Nanfang Hospital),Guangzhou 511300,China)

机构地区：[1]广州市增城区中心医院(南方医院增城分院)神经内科,广东广州511300 [2]南方医科大学南方医院神经内科,广东广州510515 [3]广州市增城区中心医院(南方医院增城分院)药剂科,广东广州511300

出　　处：《药物评价研究》2022年第9期1822-1829,共8页Drug Evaluation Research

基　　金：2021年广东省医学科研基金(B2022009)。

摘　　要：目的观察栀子苷对脑缺血再灌注损伤(CIRI)模型大鼠的神经保护作用及对胰高血糖素样肽-1受体(GLP-1R)/蛋白激酶B(Akt)信号通路的影响。方法50只SD大鼠随机分为假手术组,模型组,栀子苷低、高剂量组及尼莫地平片组,每组10只。采用线栓法制备CIRI大鼠模型,缺血2 h再灌注24 h后,栀子苷低、高剂量组大鼠分别ig给予10、40 mg·kg^(-1)的栀子苷,尼莫地平片组大鼠ig给予8.1 mg·kg^(-1)尼莫地平,假手术组和模型组大鼠ig等体积生理盐水,每天1次,连续7 d。造模后及给药结束后,分别对各组大鼠进行神经功能缺损评分;给药结束后,采用酶联免疫吸附法(ELISA)检测血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)及白细胞介素-6(IL-6)水平,苏木素-伊红(HE)染色检测脑组织病理学变化,尼氏染色检测神经元与尼氏小体变化,免疫组化染色检测脑组织B淋巴细胞瘤-2(Bcl-2)和Bcl-2关联X蛋白(Bax)表达,蛋白质印迹法(Western blotting)检测脑组织细胞色素C(Cyt-C)、半胱氨酸蛋白酶-3(Caspase-3)、半胱氨酸蛋白酶-9(Caspase-9)、胰高血糖素样肽-1受体(GLP-1R)及磷酸化蛋白激酶B(p-Akt)蛋白表达水平。结果与假手术组比较,模型组大鼠神经功能缺损评分显著升高(P<0.05),血清中TNF-α、IL-1β、IL-6水平显著升高(P<0.05);皮质细胞间质疏松、增宽,有大量神经元细胞质萎缩和细胞核损伤;脑神经元皱缩,尼氏小体变小,数目明显减少。大鼠脑组织Bcl-2表达显著降低(P<0.05),Bax表达显著升高(P<0.05);脑组织中Cyt-C、Caspase-3、Caspase-9蛋白表达水平显著升高(P<0.05),GLP-1R和p-Akt蛋白表达水平显著降低(P<0.05)。与模型组比较,栀子苷高剂量组和尼莫地平片组大鼠神经功能缺损评分均显著降低(P<0.05),血清中TNF-α、IL-1β、IL-6水平显著降低(P<0.05),皮质细胞较为整齐,神经元细胞和尼氏小体损伤减小,Bcl-2表达显著升高(P<0.05),Bax表达显著降低(P<0.05),Objective To investigate effect of geniposide on cerebral ischemia-reperfusion injury(CIRI)and its effects on glucagon like peptide-1 receptor(GLP-1R)/protein kinase B(Akt)signaling pathway.Methods Fifty SD rats were randomly divided into sham operation group,model group,low-dose geniposide group,high-dose geniposide group,and nimodipine tablet group,with 10 rats in each group.The thread embolism method was used to establish the CIRI model.After 2 h of ischemia and 24 h of reperfusion,the rats in low-dose and high-dose geniposide groups were given 10 mg·kg^(-1) and 40 mg·kg^(-1) geniposide by ig,respectively,rats in the nimodipine tablet group were given 8.1 mg·kg^(-1) nimodipine ig,rats in the sham operation group and model group were given an equal volume of normal saline,once a day for seven consecutive days.After modeling and administration,the neurological deficits of rats in each group were scored.Enzyme linked immunosorbent assay(ELISA)was used to detect the content of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)in serum.Hematoxylin-eosin(HE)staining was used to detect brain histopathological changes.Nissl staining was used to detect changes in neurons and Nissl bodies.Immunohistochemical staining was used to detect the expression of B lymphocyte tumor-2(Bcl-2)and Bcl-2 associated X protein(Bax)in brain tissue.Western blotting was used to detect cytochrome C(Cyt-C),Caspase-3,and Caspase-9,glucagon-like peptide-1 receptor(GLP-1R)and phosphorylated protein kinase B(p-Akt)protein expression levels.Results Compared with the sham operation group,the neurological deficit score of the model group was significantly higher(P<0.05),the level of TNF-α,IL-1β,and IL-6 increased significantly(P<0.05),cortical intercellular matrix was loose and widened,and there were a large number of neuronal cytoplasmic atrophy and nuclear damage,the brain neurons shrink,the Nissl body became smaller,and the number decreased significantly,the expression of Bcl-2 was significantly decreased(P<0.05

关 键 词：栀子苷 脑缺血再灌注损伤 神经元凋亡 胰高血糖素样肽-1受体(GLP-1R) 蛋白激酶B(Akt) 

分 类 号：R285.5[医药卫生—中药学]