摘要
目的:观察黄芪百合颗粒对高原低氧模型大鼠缺氧诱导因子-1α(HIF-1α)/核转录因子-κB(NF-κB)/NOD样受体热蛋白结构域相关蛋白3(NLRP3)信号通路的影响。方法:60只SPF级SD雄性大鼠随机分为正常组、模型组、地塞米松组(5 mg·kg^(-1))、黄芪百合颗粒高、中、低剂量组(4.1、2.05、1.025 g·kg^(-1))。其中各中药组连续灌胃给药14 d,1次/d,连续3 d腹腔注射地塞米松作为阳性药物组;第15天将模型组、地塞米松组、黄芪百合颗粒高、中、低剂量组至动物低压模拟舱中模拟高海拔低压低氧环境进行暴露,持续暴露3 d,腹主动脉采血并分离血清,处死后摘取脑组织;苏木素-伊红(HE)染色法观察脑组织病理变化;酶联免疫吸附测定法(ELISA)检测大鼠血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和白细胞介素-1β(IL-1β)的含量;蛋白免疫印迹法(Western blot)检测各组大鼠HIF-1α、NLRP3、磷酸化NF-κB(p-NF-κB)、NF-κB、消皮素D(GSDMD)和胱天蛋白酶-1(Caspase-1)蛋白的表达水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测各组大鼠HIF-1α、NLRP3、NF-κB p65、GSDMD和Caspase-1 mRNA的表达水平。结果:HE结果显示,与正常组比较,模型组大鼠脑组织病理切片可见锥体细胞排列松散且分布紊乱,大小不一;与模型组比较,地塞米松组及黄芪百合颗粒高、中剂量组锥体细胞病理改变减轻。ELISA结果显示,与正常组比较,模型组大鼠血清中TNF-α、IL-6和IL-1β的含量显著升高(P<0.01);与模型组比较,地塞米松组及黄芪百合颗粒高、中剂量组大鼠血清中TNF-α、IL-6和IL-1β的含量明显降低(P<0.05,P<0.01)。Western blot结果显示,与正常组比较,模型组大鼠脑组织中HIF-1α、NLRP3、p-NF-κB p65、GSDMD、Caspase-1蛋白相对表达水平均显著升高(P<0.01);与模型组比较,地塞米松组及黄芪百合颗粒高剂量组大鼠脑组织中HIF-1α、NLRP3、p-NF-κB p65、GSDMD、Caspase-1蛋白相
Objective:To observe the effect of Huangqi Baihe granules on the hypoxia-inducible factor 1α(HIF-1α)/nuclear factor-κB(NF-κB)/NOD-like receptor hot protein domain related protein 3(NLRP3)signaling pathway in a rat model of high altitude hypoxia.Method:Sixty male SPF SD rats were randomly divided into blank group,model group,dexamethasone group(5 mg·kg^(-1)),and high,middle,and low-dose groups of Huangqi Baihe granules(4.1,2.05,1.025 g·kg^(-1)).Among them,each Chinese medicine group was administrated orally for continuously 14 d,once a day,and the dexamethasone group was injected intraperitoneally for continuously 3 d as the positive control group.On the 15th d,the model group,dexamethasone group,and high,middle,and low dose groups of Huangqi Baihe granules were exposed to the simulated high altitude,low pressure,and low oxygen environment in the animal low-pressure simulation cabin,and the exposure lasted for 3 d.Blood was collected from the abdominal aorta and serum was separated,and the brain tissue was taken after being killed.Hematoxylin-eosin(HE)staining was used to observe the pathological changes in brain tissue.Enzyme-linked immunosorbent assay(ELISA)was used to detect the content of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and interleukin-1β(IL-1β)in rat serum.Western blot was used to detect HIF-1α,NLRP3,phosphorylated nuclear factor-κB(p-NF-κB),NF-κB,desquamation D(GSDMD),and cysteine aspartate-specitis protein-1(Caspase-1)in rats of each group.The mRNA expression levels of HIF-1α,NLRP3,NF-κB p65,GSDMD,and Caspase-1 were detected by real-time quantitative polymerase chain reaction(Real-time PCR).Result:The results of HE staining showed that as compared with the normal group,the pathological sections of brain tissues in the model group showed that pyramidal cells were loosely arranged and distributed in disorder,with different sizes.Compared with the model group,the pathological changes in pyramidal cells in the dexamethasone group and high and middle-dose groups of Huangqi Baih
作者
曾元丁
苏韫
龚红霞
曹旺杰
刘永琦
黄勇
冷光现
李莉霞
ZENG Yuanding;SU Yun;GONG Hongxia;CAO Wangjie;LIU Yongqi;HUANG Yong;LENG Guangxian;LI Lixia(School of Basic Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,China;Gansu University Key Laboratory for Molecular Medicine and Chinese Medicine Prevention and Treatment of Major Diseases,Lanzhou 730000,China;Lanzhou University Second Hospital,Lanzhou 730030,China;Tianshui Hospital of Integrated Traditional Chinese and Western Medicine,Tianshui 741000,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第10期134-141,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
甘肃省自然科学基金项目(21JR11RA144)
甘肃省高等学校青年博士基金项目(2022QB-102)
兰州市科技发展计划项目(2022-2-106)
甘肃中医药大学研究生创新创业基金项目(2022CX35)。
