NLRP3 inflammasome activation in gestational diabetes mellitus placentas is associated with hydrogen sulfide synthetase deficiency

Wu,Wei; Tan,Qing-Ying; Xi,Fang-Fang; Ruan,Yun; Wang,Jing; Luo,Qiong; Dou,Xiao-Bing; Hu,Tian-Xiao

doi:10.3892/etm.2021.11017

NLRP3 inflammasome activation in gestational diabetes mellitus placentas is associated with hydrogen sulfide synthetase deficiency

Authors:
- Wei Wu
- Qing-Ying Tan
- Fang-Fang Xi
- Yun Ruan
- Jing Wang
- Qiong Luo
- Xiao-Bing Dou
- Tian-Xiao Hu
View Affiliations

Affiliations: Department of Obstetrics, Women's Hospital School of Medicine Zhejiang University, Hangzhou, Zhejiang 310006, P.R. China, Department of Endocrinology, Chinese PLA 903rd Hospital (Former Chinese PLA 117th Hospital), Hangzhou, Zhejiang 310013, P.R. China, School of Life Sciences, Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310053, P.R. China
Published online on: November 30, 2021 https://doi.org/10.3892/etm.2021.11017
Article Number: 94
Copyright: © Wu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The placenta may play a key role in the activation of inflammation and initiation of insulin resistance (IR) during gestational diabetes mellitus (GDM) pathogenesis. Interleukin (IL)‑1β and IL‑18, regulated by NLR family pyrin domain containing‑3 (NLRP3) inflammasome, are important inflammatory cytokines in the initiation of maternal IR during GDM. However, the mechanism responsible for the regulatory of NLRP3 inflammasome in placenta remains unknown. Hydrogen sulfide (H₂S) exerts anti‑inflammatory function partially via suppressing the activation of the NLPR3 inflammasome. The present study aimed to investigate the role of NLRP3 inflammasome, H₂S synthetase cystathionine‑γ‑lyase (CSE) and cystathionine‑β‑synthetase (CBS) in placenta in the pathogenesis of GDM. Clinical placenta samples were collected from pregnant women with GDM (n=16) and healthy pregnant women at term (n=16). Western blot analysis was performed to detect the protein expression levels of NLRP3, cleaved caspase‑1, CBS and CSE in the placenta samples. Pearson's correlation analysis was performed to assess the correlation between NLRP3 inflammasome and H₂S synthetase. Human placental cells were cultured and treated with different concentrations of NaHS (0, 10, 25 and 50 nmol/l) or L‑cysteine (0, 0.25, 0.50 and 1.00 mmol/l). In addition, western blot analysis was performed to detect the protein expression levels of NLRP3 and cleaved caspase‑1, while ELISA was performed to measure the production of IL‑1β and IL‑18 in the culture media. The results demonstrated that the expression levels of NLRP3 and cleaved caspase‑1 increased, while the expression levels of CBS and CSE decreased in the placenta samples. In addition, the expression levels of NLRP3 and cleaved caspase‑1 were inversely correlated with the expression levels of CBS and CSE. Notably, NaHS and L‑cysteine significantly suppressed the expression levels of NLRP3 and cleaved caspase‑1, and the production of IL‑1 and IL‑18 in human placental cells. Taken together, the results of the present study suggest that H₂S synthetase deficiency in placenta may contribute to excessive activation of NLRP3 inflammasome in GDM.

View Figures

View References

1	Catalano PM: Trying to understand gestational diabetes. Diabet Med. 31:273–281. 2014.PubMed/NCBI View Article : Google Scholar
2	Yan B, Yu Y, Lin M, Li Z, Wang L, Huang P, Song H, Shi X, Yang S, Li X, et al: High, but stable, trend in the prevalence of gestational diabetes mellitus: A population-based study in Xiamen, China. J Diabetes Investig. 10:1358–1364. 2019.PubMed/NCBI View Article : Google Scholar
3	Wang M, Hu RY, Gong WW, Pan J, Fei FR, Wang H, Zhou XY, Zhong JM and Yu M: Trends in prevalence of gestational diabetes mellitus in Zhejiang Province, China, 2016-2018. Nutr Metab (Lond). 18(12)2021.PubMed/NCBI View Article : Google Scholar
4	Gualdani E, Di Cianni G, Seghieri M, Francesconi P and Seghieri G: Pregnancy outcomes and maternal characteristics in women with pregestational and gestational diabetes: A retrospective study on 206,917 singleton live births. Acta Diabetol. 58:1169–1176. 2021.PubMed/NCBI View Article : Google Scholar
5	de Sousa RAL, de Lima EV, da Silva TP, de Souza RV, Figueiredo CP, Passos GF and Clarke JR: Late Cognitive Consequences of Gestational Diabetes to the Offspring, in a New Mouse Model. Mol Neurobiol. 56:7754–7764. 2019.PubMed/NCBI View Article : Google Scholar
6	Kajantie E, Osmond C and Eriksson JG: Gestational hypertension is associated with increased risk of type 2 diabetes in adult offspring: the Helsinki Birth Cohort Study. Am J Obstet Gynecol. 216:281 e281–281 e287. 2017.PubMed/NCBI View Article : Google Scholar
7	Cinkajzlová A, Anderlová K, Šimják P, Lacinová Z, Kloučková J, Kratochvílová H, Krejčí H, Pařízek A, Mráz M, Kršek M, et al: Subclinical Inflammation and Adipose Tissue Lymphocytes in Pregnant Females With Gestational Diabetes Mellitus. J Clin Endocrinol Metab. 105(dgaa528)2020.PubMed/NCBI View Article : Google Scholar
8	Olmos-Ortiz A, Flores-Espinosa P, Díaz L, Velázquez P, Ramírez-Isarraraz C and Zaga-Clavellina V: Immunoendocrine Dysregulation during Gestational Diabetes Mellitus: The Central Role of the Placenta. Int J Mol Sci. 22(8087)2021.PubMed/NCBI View Article : Google Scholar
9	Rancourt RC, Ott R, Ziska T, Schellong K, Melchior K, Henrich W and Plagemann A: Visceral Adipose Tissue Inflammatory Factors (TNF-Alpha, SOCS3) in Gestational Diabetes (GDM): Epigenetics as a Clue in GDM Pathophysiology. Int J Mol Sci. 21(479)2020.PubMed/NCBI View Article : Google Scholar
10	Keckstein S, Pritz S, Amann N, Meister S, Beyer S, Jegen M, Kuhn C, Hutter S, Knabl J, Mahner S, et al: Sex Specific Expression of Interleukin 7, 8 and 15 in Placentas of Women with Gestational Diabetes. Int J Mol Sci. 21(8026)2020.PubMed/NCBI View Article : Google Scholar
11	Tsiotra PC, Halvatsiotis P, Patsouras K, Maratou E, Salamalekis G, Raptis SA, Dimitriadis G and Boutati E: Circulating adipokines and mRNA expression in adipose tissue and the placenta in women with gestational diabetes mellitus. Peptides. 101:157–166. 2018.PubMed/NCBI View Article : Google Scholar
12	Simpson S, Smith L and Bowe J: Placental peptides regulating islet adaptation to pregnancy: Clinical potential in gestational diabetes mellitus. Curr Opin Pharmacol. 43:59–65. 2018.PubMed/NCBI View Article : Google Scholar
13	Hill DJ: Placental control of metabolic adaptations in the mother for an optimal pregnancy outcome. What goes wrong in gestational diabetes? Placenta. 69:162–168. 2018.PubMed/NCBI View Article : Google Scholar
14	Ngala RA, Fondjo LA, Gmagna P, Ghartey FN and Awe MA: Placental peptides metabolism and maternal factors as predictors of risk of gestational diabetes in pregnant women. A case-control study. PLoS One. 12(e0181613)2017.PubMed/NCBI View Article : Google Scholar
15	Skajaa GO, Fuglsang J, Knorr S, Møller N, Ovesen P and Kampmann U: Changes in insulin sensitivity and insulin secretion during pregnancy and post partum in women with gestational diabetes. BMJ Open Diabetes Res Care. 8(e001728)2020.PubMed/NCBI View Article : Google Scholar
16	Waters TP, Kim SY, Sharma AJ, Schnellinger P, Bobo JK, Woodruff RT, Cubbins LA, Haghiac M, Minium J, Presley L, et al: Longitudinal changes in glucose metabolism in women with gestational diabetes, from late pregnancy to the postpartum period. Diabetologia. 63:385–394. 2020.PubMed/NCBI View Article : Google Scholar
17	Liu T, Deng JM, Liu YL, Chang L and Jiang YM: The relationship between gestational diabetes mellitus and interleukin 1beta gene polymorphisms in southwest of China. Medicine (Baltimore). 99(e22679)2020.PubMed/NCBI View Article : Google Scholar
18	Fatima SS, Alam F, Chaudhry B and Khan TA: Elevated levels of chemerin, leptin, and interleukin-18 in gestational diabetes mellitus. J Matern Fetal Neonatal Med. 30:1023–1028. 2017.PubMed/NCBI View Article : Google Scholar
19	Gomes CP, Torloni MR, Gueuvoghlanian-Silva BY, Alexandre SM, Mattar R and Daher S: Cytokine levels in gestational diabetes mellitus: A systematic review of the literature. Am J Reprod Immunol. 69:545–557. 2013.PubMed/NCBI View Article : Google Scholar
20	Schulze F, Wehner J, Kratschmar DV, Makshana V, Meier DT, Häuselmann SP, Dalmas E, Thienel C, Dror E, Wiedemann SJ, et al: Inhibition of IL-1beta improves Glycaemia in a Mouse Model for Gestational Diabetes. Sci Rep. 10(3035)2020.PubMed/NCBI View Article : Google Scholar
21	Zhou F, Li C and Zhang SY: NLRP3 inflammasome: A new therapeutic target for high-risk reproductive disorders? Chin Med J (Engl). 134:20–27. 2020.PubMed/NCBI View Article : Google Scholar
22	Fang X, Wang Y, Zhang Y, Li Y, Kwak-Kim J and Wu L: NLRP3 Inflammasome and Its Critical Role in Gynecological Disorders and Obstetrical Complications. Front Immunol. 11(555826)2021.PubMed/NCBI View Article : Google Scholar
23	Gora IM, Ciechanowska A and Ladyzynski P: NLRP3 Inflammasome at the Interface of Inflammation, Endothelial Dysfunction, and Type 2 Diabetes. Cells. 10(10)2021.PubMed/NCBI View Article : Google Scholar
24	Yu ZW, Zhang J, Li X, Wang Y, Fu YH and Gao XY: A new research hot spot: The role of NLRP3 inflammasome activation, a key step in pyroptosis, in diabetes and diabetic complications. Life Sci. 240(117138)2020.PubMed/NCBI View Article : Google Scholar
25	Mastrocola R, Aragno M, Alloatti G, Collino M, Penna C and Pagliaro P: Metaflammation: Tissue-Specific Alterations of the NLRP3 Inflammasome Platform in Metabolic Syndrome. Curr Med Chem. 25:1294–1310. 2018.PubMed/NCBI View Article : Google Scholar
26	Zhang R, Zhang X, Xing B, Zhao J, Zhang P, Shi D and Yang F: Astragaloside IV attenuates gestational diabetes mellitus via targeting NLRP3 inflammasome in genetic mice. Reprod Biol Endocrinol. 17(77)2019.PubMed/NCBI View Article : Google Scholar
27	Chen CQ, Xin H and Zhu YZ: Hydrogen sulfide: Third gaseous transmitter, but with great pharmacological potential. Acta Pharmacol Sin. 28:1709–1716. 2007.PubMed/NCBI View Article : Google Scholar
28	Tang C, Li X and Du J: Hydrogen sulfide as a new endogenous gaseous transmitter in the cardiovascular system. Curr Vasc Pharmacol. 4:17–22. 2006.PubMed/NCBI View Article : Google Scholar
29	Hu TX, Zhang NN, Ruan Y, Tan QY and Wang J: Hydrogen sulfide modulates high glucose-induced NLRP3 inflammasome activation in 3T3-L1 adipocytes. Exp Ther Med. 19:771–776. 2020.PubMed/NCBI View Article : Google Scholar
30	Teng Y, Xuan S, Jiang M, Tian L, Tian J and Chang Q: Expression of H₂S in Gestational Diabetes Mellitus and Correlation Analysis with Inflammatory Markers IL-6 and TNF-α. J Diabetes Res. 2020(3085840)2020.PubMed/NCBI View Article : Google Scholar
31	Renga B: Hydrogen sulfide generation in mammals: The molecular biology of cystathionine-β- synthase (CBS) and cystathionine-γ-lyase (CSE). Inflamm Allergy Drug Targets. 10:85–91. 2011.PubMed/NCBI View Article : Google Scholar
32	Hu T, Wang G, Zhu Z, Huang Y, Gu H and Ni X: Increased ADAM10 expression in preeclamptic placentas is associated with decreased expression of hydrogen sulfide production enzymes. Placenta. 36:947–950. 2015.PubMed/NCBI View Article : Google Scholar
33	Sun Q, Chen Z, He P, Li Y, Ding X, Huang Y, Gu H and Ni X: Reduced Expression of Hydrogen Sulfide-Generating Enzymes Down-Regulates 15-Hydroxyprostaglandin Dehydrogenase in Chorion during Term and Preterm Labor. Am J Pathol. 188:63–71. 2018.PubMed/NCBI View Article : Google Scholar
34	Lu L, Kingdom J, Burton GJ and Cindrova-Davies T: Placental Stem Villus Arterial Remodeling Associated with Reduced Hydrogen Sulfide Synthesis Contributes to Human Fetal Growth Restriction. Am J Pathol. 187:908–920. 2017.PubMed/NCBI View Article : Google Scholar
35	Ganguly E, Hula N, Spaans F, Cooke CM, Davidge ST and Ganguly E: Placenta-targeted treatment strategies: An opportunity to impact fetal development and improve offspring health later in life. Pharmacol Res. 157(104836)2020.PubMed/NCBI View Article : Google Scholar
36	Tsai K, Tullis B, Jensen T, Graff T, Reynolds P and Arroyo J: Differential expression of mTOR related molecules in the placenta from gestational diabetes mellitus (GDM), intrauterine growth restriction (IUGR) and preeclampsia patients. Reprod Biol. 21(100503)2021.PubMed/NCBI View Article : Google Scholar
37	Awamleh Z, Butcher DT, Hanley A, Retnakaran R, Haertle L, Haaf T, Hamilton J and Weksberg R: Exposure to Gestational Diabetes Mellitus (GDM) alters DNA methylation in placenta and fetal cord blood. Diabetes Res Clin Pract. 174(108690)2021.PubMed/NCBI View Article : Google Scholar
38	Sarina Li DF, Feng ZQ, Du J, Zhao WH, Huang N, Jia JC, Wu ZY, Alamusi Wang YY, Ji XL and Yu L: Mechanism of Placenta Damage in Gestational Diabetes Mellitus by Investigating TXNIP of Patient Samples and Gene Functional Research in Cell Line. Diabetes Ther. 10:2265–2288. 2019.PubMed/NCBI View Article : Google Scholar
39	Dinarello CA: Overview of the IL-1 family in innate inflammation and acquired immunity. Immunol Rev. 281:8–27. 2018.PubMed/NCBI View Article : Google Scholar
40	Ballak DB, Stienstra R, Tack CJ, Dinarello CA and van Diepen JA: IL-1 family members in the pathogenesis and treatment of metabolic disease: Focus on adipose tissue inflammation and insulin resistance. Cytokine. 75:280–290. 2015.PubMed/NCBI View Article : Google Scholar
41	Zochio GP, Possomato-Vieira JS, Chimini JS, da Silva MLS and Dias-Junior CA: Effects of fast versus slow-releasing hydrogen sulfide donors in hypertension in pregnancy and fetoplacental growth restriction. Naunyn Schmiedebergs Arch Pharmacol. 392:1561–1568. 2019.PubMed/NCBI View Article : Google Scholar
42	You X, Chen Z, Zhao H, Xu C, Liu W, Sun Q, He P, Gu H and Ni X: Endogenous hydrogen sulfide contributes to uterine quiescence during pregnancy. Reproduction. 153:535–543. 2017.PubMed/NCBI View Article : Google Scholar
43	Jia Q, Mehmood S, Liu X, Ma S and Yang R: Hydrogen sulfide mitigates myocardial inflammation by inhibiting nucleotide-binding oligomerization domain-like receptor protein 3 inflammasome activation in diabetic rats. Exp Biol Med (Maywood). 245:221–230. 2020.PubMed/NCBI View Article : Google Scholar
44	Zheng Q, Pan L and Ji Y: H 2S protects against diabetes-accelerated atherosclerosis by preventing the activation of NLRP3 inflammasome. J Biomed Res. 34:94–102. 2019.PubMed/NCBI View Article : Google Scholar
45	Su Y, Wang Y, Liu M and Chen H: Hydrogen sulfide attenuates renal I/R-induced activation of the inflammatory response and apoptosis via regulating Nrf2-mediated NLRP3 signaling pathway inhibition. Mol Med Rep. 24(518)2021.PubMed/NCBI View Article : Google Scholar
46	Nguyen TTP, Kim DY, Lee YG, Lee YS, Truong XT, Lee JH, Song DK, Kwon TK, Park SH, Jung CH, et al: SREBP-1c impairs ULK1 sulfhydration-mediated autophagic flux to promote hepatic steatosis in high-fat-diet-fed mice. Mol Cell. 81:3820–3832.e7. 2021.PubMed/NCBI View Article : Google Scholar
47	Peh MT, Anwar AB, Ng DS, Atan MS, Kumar SD and Moore PK: Effect of feeding a high fat diet on hydrogen sulfide (H₂S) metabolism in the mouse. Nitric Oxide. 41:138–145. 2014.PubMed/NCBI View Article : Google Scholar
48	Moreira AM, Grisote SA, Francescato HDC, Coimbra TM, Elias LLK, Antunes-Rodrigues J and Ruginsk SG: Effects of endogenous H₂S production inhibition on the homeostatic responses induced by acute high-salt diet consumption. Mol Cell Biochem. 476:715–725. 2021.PubMed/NCBI View Article : Google Scholar
49	Zheng W and Liu C: The cystathionine γ-lyase/hydrogen sulfide pathway mediates the trimetazidine-induced protection of H9c2 cells against hypoxia/reoxygenation-induced apoptosis and oxidative stress. Anatol J Cardiol. 22:102–111. 2019.PubMed/NCBI View Article : Google Scholar
50	Bibli SI and Fleming I: Oxidative Post-Translational Modifications: A Focus on Cysteine S-Sulfhydration and the Regulation of Endothelial Fitness. Antioxid Redox Signal: Sep 29, 2021 (Epub ahead of print). doi: 10.1089/ars.2021.0162.
51	Manna P and Jain SK: Vitamin D up-regulates glucose transporter 4 (GLUT4) translocation and glucose utilization mediated by cystathionine-γ-lyase (CSE) activation and H₂S formation in 3T3L1 adipocytes. J Biol Chem. 287:42324–42332. 2012.PubMed/NCBI View Article : Google Scholar
52	Brown J, Alwan NA, West J, Brown S, McKinlay CJ, Farrar D and Crowther CA: Lifestyle interventions for the treatment of women with gestational diabetes. Cochrane Database Syst Rev. 5(CD011970)2017.PubMed/NCBI View Article : Google Scholar
53	Jiang XC, Liang ZD, Chen DL, Jia JP, Hu JR and Hu L: Correlation of Homocysteine, AHSG, CRP with Insulin Resistance, 25-(OH)2-VitD, Blood Lipids in Gestational Diabetes Patients. Clin Lab. 67(67)2021.PubMed/NCBI View Article : Google Scholar
54	Chen W, Li Y, Gao B, Li J, Zheng M and Chen X: Serum 25-hydroxyvitamin D levels in relation to lipids and clinical outcomes in pregnant women with gestational diabetes mellitus: An observational cohort study. BMJ Open. 10(e039905)2020.PubMed/NCBI View Article : Google Scholar
55	Akarsu S, Bagirzade M, Omeroglu S and Büke B: Placental vascularization and apoptosis in Type-1 and gestational DM. J Matern Fetal Neonatal Med. 30:1045–1050. 2017.PubMed/NCBI View Article : Google Scholar
56	Kasture V, Sahay A and Joshi S: Cell death mechanisms and their roles in pregnancy related disorders. Adv Protein Chem Struct Biol. 126:195–225. 2021.PubMed/NCBI View Article : Google Scholar