Huangdi Anxiao Capsules-containing serum protects cell model from cognitive dysfunction in diabetes via inhibiting NLRP3-mediated pyroptosis.
10.19540/j.cnki.cjcmm.20230614.702
- Author:
Xiao-Juan WANG
1
;
Yu-Lu WANG
1
;
Nan SHAO
1
;
Ting YE
1
;
Shu YE
1
;
Hua-Wu GAO
1
;
Yan WANG
1
Author Information
1. School of Integrated Chinese and Western Medicine, Anhui University of Chinese Medicine Hefei 230012, China Anhui Provincial Medical Basic Research and Innovation Center for the Prevention and Treatment of Neurodegenerative Diseases by Integrating Traditional Chinese and Western Medicine Hefei 230012, China.
- Publication Type:Journal Article
- Keywords:
Huangdi Anxiao Capsules-containing serum;
NLRP3 inflammasome;
PC12 cells;
pyroptosis
- MeSH:
Rats;
Animals;
NLR Family, Pyrin Domain-Containing 3 Protein/metabolism*;
Interleukin-18;
Pyroptosis/physiology*;
Diabetes Mellitus;
Caspases;
Glucose;
RNA, Messenger
- From:
China Journal of Chinese Materia Medica
2023;48(19):5315-5325
- CountryChina
- Language:Chinese
-
Abstract:
This study aims to investigate the effects and the molecular mechanism of Huangdi Anxiao Capsules(HDAX)-containing serum in protecting the rat adrenal pheochromocytoma(PC12) cells from diabetes-associated cognitive dysfunction induced by high glucose and whether the mechanism is related to the regulation of NOD-like receptor thermal protein domain associated protein 3(NLRP3)-mediated pyroptosis. The PC12 cell model of diabetes-associated cognitive dysfunction induced by high glucose was established and mcc950 was used to inhibit NLRP3. PC12 cells were randomized into control, model, HDAX-containing serum, mcc950, and HDAX-containing serum+mcc950 groups. Methyl thiazolyl tetrazolium(MTT) assay was employed to determine the viability, and Hoechst 33258/PI staining to detect pyroptosis of PC12 cells. Enzyme-linked immunosorbent assay(ELISA) was employed to measure the levels of interleukin-1 beta(IL-1β) and IL-18. Western blot was employed to determine the protein levels of postsynaptic density protein 95(PSD-95), NLRP3, apoptosis-associated speck-like protein containing a CARD(ASC), gasdermin D(GSDMD), GSDMD-N, and cleaved cysteinyl aspartate specific proteinase-1(caspase-1), and RT-PCR to determine the mRNA levels of NLRP3, ASC, GSDMD, and caspase-1. The immunofluorescence assay was adopted to measure the levels and distribution of NLRP3 and GSDMD-N in PC12 cells. Compared with the control group, the model group showed decreased cell proliferation, increased PI positive rate, down-regulated protein level of PSD-95, up-regulated protein levels of NLRP3, ASC, GSDMD-N, GSDMD, and cleaved caspase-1, up-regulated mRNA levels of NLRP3, ASC, GSDMD, and caspase-1, and elevated levels of IL-1β and IL-18. Compared with the model group, HDAX-containing serum, mcc950, and the combination of them improved cell survival rate and morphology, decreased the PI positive rate, down-regulated the protein levels of NLRP3, ASC, GSDMD-N, GSDMD, and cleaved caspase-1 and the mRNA levels of NLRP3, ASC, GSDMD, and caspase-1, and promoted the secretion of IL-1β and IL-18. The findings demonstrated that HDAX-containing serum can inhibit the pyroptosis-mediated by NLRP3 and protect PC12 cells from the cognitive dysfunction induced by high glucose.
