Li Na, Mu Ya-Ping, Liu Chun-Ying, Wang Yang, Li Xiao-Feng, Wang Xue-Wei
Integrated Traditional Chinese and Western Medicine, Liaoning University of Traditional Chinese Medicine, Shenyang 110847, China.
Zhongguo Dang Dai Er Ke Za Zhi. 2021 Apr;23(4):402-409. doi: 10.7499/j.issn.1008-8830.2010053.
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).
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).
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 ( < 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 ( < 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 ( < 0.05).
AS-IV may alleviate HIBD in neonatal rats by inhibiting the expression of NLRP3, GSDMD, caspase-1, and IL-1β.
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。
选取24只7日龄Sprague-Dawley大鼠，随机分为假手术组、HIBD组和AS-IV治疗组，每组8只。造模24小时后，取脑组织进行苏木精-伊红染色、yo-PRO-1染色和EthD-2免疫荧光染色，观察AS-IV在体内的脑保护作用。采用HT22细胞制备氧糖剥夺（OGD）模型，并建立AS-IV浓度梯度（50-400μmol/L）。采用CCK-8法检测HT22细胞活力。采用RT-PCR和蛋白质印迹法观察不同浓度AS-IV对NLRP3、gasdermin D（GSDMD）、半胱天冬酶-1（caspase-1）和白细胞介素-1β（IL-1β）mRNA及蛋白表达的影响。
Yo-Pro-1和EthD-2染色显示，与假手术组相比，HIBD组焦亡细胞增多，有少量坏死细胞，而AS-IV组焦亡和坏死细胞均减少。与假手术组相比，HIBD组NLRP3、IL-1β、caspase-1和GSDMD蛋白表达水平显著升高（<0.05）。与HIBD组相比，AS-IV组NLRP3、caspase-1和GSDMD蛋白表达水平显著降低（<0.05）。HT22细胞实验显示，与OGD组相比，AS-IV组NLRP3、GSDMD、caspase-1和IL-1β的mRNA和蛋白表达均受到抑制，200μmol/L浓度时治疗效果最佳（<0.05）。
AS-IV可能通过抑制NLRP3、GSDMD、caspase-1和IL-1β的表达减轻新生大鼠HIBD。
