文献类型：期刊文献

中文题名：基于HIF-1α/NF-κB/NLRP3信号通路探讨黄芪百合颗粒对高原低氧模型大鼠急性脑损伤的保护作用

英文题名：Protective Effect of Huangqi Baihe Granules on Acute Brain Injury in Rats with High Altitude Hypoxia Based on HIF-1α/NF-κB/NLRP3 Signal Pathway

作者：曾元丁[1,2];苏韫[1,2];龚红霞[1,2];曹旺杰[1,2];刘永琦[1,2];黄勇[1,2];冷光现[3];李莉霞[4]

第一作者：曾元丁

机构：[1]甘肃中医药大学基础医学院,兰州730000;[2]甘肃省高校重大疾病分子医学与中医药防治研究省级重点实验室,兰州730000;[3]兰州大学第二医院,兰州730030;[4]天水市中西医结合医院,甘肃天水741000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）

年份：2023

卷号：29

期号：10

起止页码：134

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

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)。

语种：中文

中文关键词：黄芪百合颗粒;高原低氧;脑损伤;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;炎症反应

外文关键词：Huangqi Baihe granules;high altitude hypoxia;brain injury;hypoxia-inducible factor 1α(HIF-1α)/nuclear factor-κB(NF-κB)/NOD-like receptor hot protein domain related protein 3(NLRP3)signaling pathway;inflammatory reaction

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蛋白相对表达均明显降低(P<0.05,P<0.01),黄芪百合颗粒中剂量组大鼠脑组织中HIF-1α、NLRP3、p-NF-κB p65、Caspase-1蛋白相对表达均显著降低(P<0.01),黄芪百合颗粒低剂量组大鼠脑组织中HIF-1α蛋白相对表达降低(P<0.05)。Real-time PCR分析显示,与正常组比较,模型组大鼠脑组织HIF-1α、NLRP3、NF-κB p65、GSDMD和Caspase-1mRNA表达水平显著增加(P<0.01);与模型组比较,地塞米松组大鼠脑组织中HIF-1α、NLRP3、NF-κBp65、GSDMD和Caspase-1mRNA表达水平显著降低(P<0.01);黄芪百合颗粒高剂量组大鼠脑组织中HIF-1α、NF-κBp65、GSDMD和Caspase-1 mRNA表达水平显著降低(P<0.01);黄芪百合颗粒中剂量组大鼠脑组织中HIF-1α、NLRP3和Caspase-1 mRNA表达水平降低(P<0.05,P<0.01)。结论:黄芪百合颗粒对低压低氧大鼠急性脑损伤的保护作用可能与HIF-1α/NF-κB/NLRP3信号通路相关。
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 Baihe granules were reduced.The results of ELISA showed that as compared with the normal group,the content of TNF-α,IL-6,and IL-1βin the serum of rats in the model group was significantly higher(P<0.01).Compared with the model group,the content of TNF-α,IL-6,and IL-1βin the serum of rats in the dexamethasone group and high and middle-dose groups of Huangqi Baihe granules decreased significantly(P<0.05,P<0.01).The results of Western blot showed that as compared with the normal group,the relative protein expression levels of HIF-1α,NLRP3,p-NF-κB p65,GSDMD,and Caspase-1 in the brain tissue of the model group were significantly higher(P<0.01).As compared with the model group,the relative expressions of HIF-1α,NLRP3,p-NF-κB p65,GSDMD,and Caspase-1 in the brain tissue of rats in the dexamethasone group and the high-dose group of Huangqi Baihe granules were significantly decreased(P<0.05,P<0.01).The relative protein expression levels of HIF-1α,NLRP3,p-NF-κB p65,and Caspase-1 in the brain tissue of rats in the middle-dose group of Huangqi Baihe granules decreased significantly(P<0.01),and the relative protein expression of HIF-1αin the brain tissue of rats in the low-dose group of Huangqi Baihe granules was reduced(P<0.05).The Real-time PCR analysis showed that as compared with the normal group,the mRNA expression levels of HIF-1α,NLRP3,NF-κB p65,GSDMD,and Caspase-1 in the brain tissue of the model group were significantly increased(P<0.01).As compared with the model group,the mRNA expression levels of HIF-1α,NLRP3,NF-κB p65,GSDMD,and Caspase-1 in the brain tissue of rats in the dexamethasone group were significantly decreased(P<0.01).The mRNA expression levels of HIF-1α,NF-κB p65,GSDMD,and Caspase-1 in the brain tissue of rats in the high-dose group of Huangqi Baihe granules decreased significantly(P<0.01).The mRNA expression levels of HIF-1α,NLRP3,and Caspase-1in the brain tissue of rats in the middle-dose group of Huangqi granules decreased(P<0.05,P<0.01).Conclusion:The protective effect of Huangqi Baihe granules on acute brain injury in low-pressure hypoxic rats may be related to the HIF-1α/NF-κB/NLRP3 signaling pathway.