[Effect of astragaloside IV on the expression of NOD-like receptor protein 3 inflammasome in neonatal rats with hypoxic-ischemic brain damage]

Zhongguo Dang Dai Er Ke Za Zhi. 2021 Apr;23(4):402-409. doi: 10.7499/j.issn.1008-8830.2010053.
[Article in Chinese]

Abstract

Objective: To study the effect of astragaloside IV (AS-IV) on NOD-like receptor protein 3 (NLRP3) inflammasome in neonatal rats with hypoxic-ischemic brain damage (HIBD).

Methods: A total of 24 Sprague-Dawley rats, aged 7 days, were randomly divided into a sham-operation group, an HIBD group, and an AS-IV treatment group, with 8 rats in each group. After 24 hours of modeling, brain tissue was collected for hematoxylin-eosin staining, yo-PRO-1 staining, and EthD-2 immunofluorescent staining in order to observe the cerebral protection effect of AS-IV in vivo. HT22 cells were used to prepare a model of oxygen-glycogen deprivation (OGD), and a concentration gradient (50-400 μmol/L) was established for AS-IV. CCK-8 assay was used to measure the viability of HT22 cells. RT-PCR and Western blot were used to observe the effect of different concentrations of AS-IV on the mRNA and protein expression of NLRP3, gasdermin D (GSDMD), caspase-1, and interleukin-1β (IL-1β).

Results: Yo-Pro-1 and EthD-2 staining showed that compared with the sham-operation group, the HIBD group had an increase in pyroptotic cells with a small number of necrotic cells, and the AS-IV group had reductions in both pyroptotic and necrotic cells. Compared with the sham-operation group, the HIBD group had significantly higher protein expression levels of NLRP3, IL-1β, caspase-1, and GSDMD (P < 0.05). Compared with the HIBD group, the AS-IV group had significant reductions in the protein expression levels of NLRP3, caspase-1, and GSDMD (P < 0.05). HT22 cell experiment showed that compared with the OGD group, the AS-IV group had inhibited mRNA and protein expression of NLRP3, GSDMD, caspase-1, and IL-1β, with the best therapeutic effect at the concentration of 200 μmol/L (P < 0.05).

Conclusions: AS-IV may alleviate HIBD in neonatal rats by inhibiting the expression of NLRP3, GSDMD, caspase-1, and IL-1β.

目的: 探讨黄芪甲苷(AS-IV)对缺氧缺血性脑损伤(HIBD)新生大鼠Nod样受体蛋白3(NLRP3)炎性小体的影响。

方法: 将24只7日龄Sprague-Dawley大鼠随机分为假手术组、HIBD组、AS-IV治疗组(AS-IV组),每组8只。各组动物造模24 h后取脑组织进行苏木精-伊红染色、yo-PRO-1和EthD-2免疫荧光染色,以观察AS-IV在体内的脑保护作用。构建HT22细胞氧糖剥夺(OGD)模型,设置AS-IV浓度梯度(50~400 μmol/L)进行干预。采用CCK8法检测HT22细胞活力,RT-PCR和Western blot法检测不同浓度AS-IV对NLRP3、Gasdermin D蛋白(GSDMD)、半胱氨酸蛋白酶-1(Caspase-1)、白细胞介素1β(IL-1β)mRNA及其蛋白表达的影响。

结果: Yo-PRO-1和EthD-2染色结果显示,与假手术组比较,HIBD组焦亡细胞增多,同时可见少量坏死细胞,AS-IV组焦亡细胞和坏死细胞均减少。HIBD组脑组织NLRP3、IL-1β、Caspase-1及GSDMD蛋白表达水平显著高于假手术组(P < 0.05);与HIBD组比较,AS-IV组NLRP3、Caspase-1及GSDMD蛋白表达水平显著降低(P < 0.05)。HT22细胞实验结果显示,与OGD组比较,AS-IV可以抑制NLRP3、GSDMD、Caspase-1、IL-1β mRNA及其蛋白表达,200 μmol/L时治疗效果最佳(P < 0.05)。

结论: AS-IV可能通过抑制NLRP3、GSDMD、Caspase-1、IL-1β的表达减轻新生大鼠HIBD。

MeSH terms

  • Animals
  • Animals, Newborn
  • Brain
  • Hypoxia-Ischemia, Brain* / drug therapy
  • Inflammasomes*
  • NLR Proteins
  • Rats
  • Rats, Sprague-Dawley
  • Saponins
  • Triterpenes

Substances

  • Inflammasomes
  • NLR Proteins
  • Saponins
  • Triterpenes
  • astragaloside A

Grants and funding

辽宁省自然科学基金计划项目(20180550290);沈阳市中青年科技创新人才支持计划项目(RC180035)