Acute glucose fluctuation promotes <em>in vitro</em> intestinal epithelial cell apoptosis and inflammation via the NOX4/ROS/JAK/STAT3 signaling pathway

Chen,Bingyu; Jia,Yuanyuan; Lu,Dongxue; Sun,Zhiguang

doi:10.3892/etm.2021.10120

Acute glucose fluctuation promotes in vitro intestinal epithelial cell apoptosis and inflammation via the NOX4/ROS/JAK/STAT3 signaling pathway

Authors:
- Bingyu Chen
- Yuanyuan Jia
- Dongxue Lu
- Zhiguang Sun
View Affiliations

Affiliations: Department of Gastroenterology, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210000, P.R. China, Department of Medical Oncology, The Second People's Hospital of Huai'an, Huaian, Jiangsu 223001, P.R. China
Published online on: April 28, 2021 https://doi.org/10.3892/etm.2021.10120
Article Number: 688
Copyright: © Chen 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

High blood glucose commonly occurs in patients with diabetes mellitus, but little is known of its effects on intestinal epithelial cells, or its associated mechanisms of action therein. In the present study, intestinal epithelial cells were assigned to five groups: i) The normal glucose (NG) group, incubated in 5.0 mmol/l glucose; ii) the constant high glucose (CHG) group, treated with 25.0 mmol/l glucose; iii) the intermittent high glucose (IHG) group, treated with alternating doses of 5.0 and 25.0 mmol/l glucose every 8 h; iv) the mannose group, cultured in 25.0 mmol/l mannose (the osmotic control); and v) the IHG glucose + GKT137831 group, pretreated with 100 nmol/l NADPH oxidase 4 (NOX4) inhibitor, GKT137831, and then exposed to IHG. TNF‑α, IL‑1 and IL‑6 levels were quantified using ELISA kits. Intestinal epithelial cell apoptosis was assessed by flow cytometry and oxidative stress was evaluated by reactive oxygen species (ROS) and malondialdehyde (MDA) detection. The expression levels of proteins associated with apoptosis and involved in the signal transduction of Janus kinase (JAK)/STAT3 pathway were assessed using western blot analysis. The results indicated that NOX4 expression was significantly higher in the CHG group than in the NG group (P<0.01), but lower than in the IHG group (P<0.001). The IHG group exhibited apoptosis and oxidative stress accompanied by the most significant increase in MDA, ROS and inflammatory cytokine levels (P<0.001), which was followed by that of the CHG group. Additionally, the IHG group exhibited reduced Bcl‑2, as well as enhanced Bax and cleaved caspase‑3 levels compared with the CHG group (P<0.001). Furthermore, the level of phosphorylated (p‑)JAK/p‑STAT3 was increased to a greater extent in the IHG group than in the CHG group (P<0.001). In conclusion, the findings of the present study indicated that CHG may trigger intestinal epithelial cell apoptosis and inflammation through the NOX4/ROS/JAK/STAT3 pathway, which may be aggravated by acute glucose fluctuation.

View Figures

View References

1	Wilson PWF, D'Agostino RB, Helen P, Lisa S and Meigs JB: Metabolic syndrome as a precursor of cardiovascular disease and type 2 diabetes mellitus. Circulation. 112:3066–3072. 2005.PubMed/NCBI View Article : Google Scholar
2	Brownlee M: Biochemistry and molecular cell biology of diabetic complications. Nature. 414:813–820. 2001.PubMed/NCBI View Article : Google Scholar
3	Papatheodorou K, Banach M, Bekiari E, Rizzo M and Edmonds M: Complications of diabetes 2017. J Diabetes Res. 2018(3086167)2018.PubMed/NCBI View Article : Google Scholar
4	Snelson M, de Pasquale C, Ekinci EI and Coughlan MT: Gut microbiome, prebiotics, intestinal permeability and diabetes complications. Best Pract Res Clin Endocrinol Metab, 2021 (Ahead of print).
5	Talley NJ, Young L, Bytzer P, Hammer J, Leemon M, Jones M and Horowitz M: Impact of chronic gastrointestinal symptoms in diabetes mellitus on health-related quality of life. Am J Gastroenterol. 96:71–76. 2001.PubMed/NCBI View Article : Google Scholar
6	Mooradian AD, Morley JE, Levine AS, Prigge WF and Gebhard RL: Abnormal intestinal permeability to sugars in diabetes mellitus. Diabetologia. 29:221–224. 1986.PubMed/NCBI View Article : Google Scholar
7	Meddings JB, Jarand J, Urbanski SJ, Hardin J and Gall DG: Increased gastrointestinal permeability is an early lesion in the spontaneously diabetic BB rat. Am J Physiol. 276:951–957. 1999.PubMed/NCBI View Article : Google Scholar
8	Neu J, Reverte CM, Mackey AD, Liboni K, Tuhacek-Tenace LM, Hatch M, Li N, Caicedo RA, Schatz DA and Atkinson M: Changes in intestinal morphology and permeability in the biobreeding rat before the onset of type 1 diabetes. J Pediatr Gastroenterol Nutr. 40:589–595. 2005.PubMed/NCBI View Article : Google Scholar
9	Chocarro-Calvo A, García-Martínez JM, Ardila-González S, De la Vieja A and García-Jiménez C: Glucose-induced β-catenin acetylation enhances Wnt signaling in cancer. Mol Cell. 49:474–486. 2013.PubMed/NCBI View Article : Google Scholar
10	Baynes JW and Thorpe SR: Role of oxidative stress in diabetic complications: A new perspective on an old paradigm. Diabetes. 43:1–9. 1999.PubMed/NCBI View Article : Google Scholar
11	Babior BM: NADPH oxidase. Curr Opin Immunol. 16:42–47. 2004.PubMed/NCBI View Article : Google Scholar
12	Ago T, Kuroda J, Kamouchi M, Sadoshima J and Kitazono T: Pathophysiological roles of NADPH oxidase/nox family proteins in the vascular system. Review and perspective. Circ J. 75:1791–1800. 2011.PubMed/NCBI View Article : Google Scholar
13	Asaba K, Tojo A, Onozato ML, Goto A, Quinn MT, Fujita T and Wilcox CS: Effects of NADPH oxidase inhibitor in diabetic nephropathy. Kidney Int. 67:1890–1898. 2005.PubMed/NCBI View Article : Google Scholar
14	Roe ND, Thomas DP and Ren J: Inhibition of NADPH oxidase alleviates experimental diabetes-induced myocardial contractile dysfunction. Diabetes Obes Metab. 13:465–473. 2011.PubMed/NCBI View Article : Google Scholar
15	Teixeira G, Szyndralewiez C, Molango S, Carnesecchi S, Heitz F, Wiesel P and Wood JM: Therapeutic potential of NADPH oxidase 1/4 inhibitors. Br J Pharmacol. 174:1667–1669. 2017.PubMed/NCBI View Article : Google Scholar
16	Wang JW, Pan YB, Cao YQ, Wang C, Jiang WD, Zhai WF and Lu JG: Loganin alleviates LPS-activated intestinal epithelial inflammation by regulating TLR4/NF-κB and JAK/STAT3 signaling pathways. Kaohsiung J Med Sci. 36:257–264. 2020.PubMed/NCBI View Article : Google Scholar
17	Hirano T, Ishihara KM and Hibi M: Roles of STAT3 in mediating the cell growth, differentiation and survival signals relayed through the IL-6 family of cytokine receptors. Oncogene. 19:2548–2556. 2000.PubMed/NCBI View Article : Google Scholar
18	Monnier L, Mas E, Ginet C, Michel F, Villon L, Cristol JP and Colette C: Activation of oxidative stress by acute glucose fluctuations compared with sustained chronic hyperglycemia in patients with type 2 diabetes. JAMA. 295:1681–1687. 2006.PubMed/NCBI View Article : Google Scholar
19	Ceriello A, Esposit K, Piconi L, Ihnat MA, Thorpe JE, Testa R, Boemi M and Giugliano D: Oscillating glucose is more deleterious to endothelial function and oxidative stress than mean glucose in normal and type 2 diabetic patients. Diabetes. 57:1349–1354. 2008.PubMed/NCBI View Article : Google Scholar
20	Lee MK, Kim IH, Choi YH and Nam TJ: A peptide from Porphyra yezoensisstimulates the proliferation of IEC-6 cells by activating the insulin-like growth factor I receptor signaling pathway. Int J Mol Med. 35:533–538. 2015.PubMed/NCBI View Article : Google Scholar
21	Shen JT, Li YS, Xia ZQ, Wen SH, Yao X, Yang WJ, Li C and Liu KX: Remifentanil preconditioning protects the small intestine against ischemia/reperfusion injury via intestinal δ- and µ-opioid receptors. Surgery. 159:548–559. 2016.PubMed/NCBI View Article : Google Scholar
22	Ying C, Wang S, Lu Y, Chen L, Mao Y, Ling H, Cheng X and Zhou X: Glucose fluctuation increased mesangial cell apoptosis related to AKT signal pathway. Arch Med Sci. 15:730–737. 2019.PubMed/NCBI View Article : Google Scholar
23	Hsieh CF, Liu CK, Lee CT, Yu LE and Wang JY: Acute glucose fluctuation impacts microglial activity, leading to inflammatory activation or self-degradation. Sci Rep. 9(840)2019.PubMed/NCBI View Article : Google Scholar
24	Quagliaro L, Piconi L, Assaloni R, Martinelli L, Motz E and Ceriello A: Intermittent high glucose enhances apoptosis related to oxidative stress in human umbilical vein endothelial cells: The Role of Protein Kinase C and NAD(P)H-Oxidase Activation. Diabetes. 52:2795–2804. 2003.PubMed/NCBI View Article : Google Scholar
25	Cho JH, Chang SA, Kwon HS, Choi YH, Ko SH, Moon SD, Yoo SJ, Song KH, Son HS, Kim HS, et al: Long-term effect of the Internet-based glucose monitoring system on HbA1c reduction and glucose stability: A 30-month follow-up study for diabetes management with a ubiquitous medical care system. Diabetes Care. 29:2625–2631. 2006.PubMed/NCBI View Article : Google Scholar
26	Liu Q, Huang QX, Lou FC, Zhang L and Hou WK, Yu S, Xu H, Wang Q, Zhang Y and Hou WK: Effects of glucose and insulin on the H9c2 (2-1) cell proliferation may be mediated through regulating glucose transporter 4 expression. Chin Med J (Engl). 126:4037–4042. 2013.PubMed/NCBI
27	Lopez R, Arumugam A, Joseph R, Monga K, Boopalan T, Agullo P, Gutierrez C, Nandy S, Subramani R, de la Rosa JM and Lakshmanaswamy R: Hyperglycemia enhances the proliferation of non-tumorigenic and malignant Mammary epithelial cells through increased leptin/IGF1R signaling and activation of AKT/mTOR. PLoS One. 8(e79708)2013.PubMed/NCBI View Article : Google Scholar
28	Ozmen A, Unek G, Kipmen-Korgun D and Korgun ET: The PI3K/Akt and MAPK-ERK1/2 pathways are altered in STZ induced diabetic rat placentas. Histol Histopathol. 29:743–756. 2013.PubMed/NCBI View Article : Google Scholar
29	Adachi T, Mori C, Sakurai K, Shihara N, Tsuda K and Yasuda K: Morphological changes and increased sucrase and isomaltase activity in small intestines of insulin-deficient and type 2 diabetic rats. Endocr J. 50:271–279. 2003.PubMed/NCBI View Article : Google Scholar
30	Rasool S, Kadla SA, Rasool V and Ganai BA: A comparative overview of general risk factors associated with the incidence of colorectal cancer. Tumor Biology. 34:2469–2476. 2013.PubMed/NCBI View Article : Google Scholar
31	Hata K, Kubota M, Shimizu M, Moriwaki H, Kuno T, Tanaka T, Hara A and Hirose Y: Monosodium glutamate-induced diabetic mice are susceptible to azoxymethane-induced colon tumorigenesis. Carcinogenesis. 33:702–707. 2012.PubMed/NCBI View Article : Google Scholar
32	King GL and Loeken MR: Hyperglycemia-induced oxidative stress in diabetic complications. Histochem Cell Biol. 122:333–338. 2004.PubMed/NCBI View Article : Google Scholar
33	Wu N, Shen H, Liu H, Wang Y, Bai Y and Han P: Acute blood glucose fluctuation enhances rat aorta endothelial cell apoptosis, oxidative stress and pro-inflammatory cytokine expression in vivo. Cardiovasc Diabetol. 15(109)2016.PubMed/NCBI View Article : Google Scholar
34	Sharma A, Tate M, Mathew G, Vince JE, Ritchie RH and Haan JB: Oxidative stress and NLRP3-inflammasome activity as significant drivers of diabetic cardiovascular complications: Therapeutic implications. Front Physiol. 9(114)2018.PubMed/NCBI View Article : Google Scholar
35	Lal MA, Brismar H, Eklöf AC and Aperia A: Role of oxidative stress in advanced glycation end product-induced mesangial cell activation. Kidney Int. 61:2006–2014. 2002.PubMed/NCBI View Article : Google Scholar
36	Zhang W, Zhao S, Li Y, Peng G and Han P: Acute blood glucose fluctuation induces myocardial apoptosis through oxidative stress and nuclear factor-ĸB activation. Cardiology. 124:11–17. 2013.PubMed/NCBI View Article : Google Scholar
37	Xue B, Wang L, Zhang Z, Wang R, Xia XX, Han PP, Cao LJ, Liu YH and Sun LQ: Puerarin may protect against Schwann cell damage induced by glucose fluctuation. J Nat Med. 71:472–481. 2017.PubMed/NCBI View Article : Google Scholar
38	Xia J, Hu S, Xu J, Hao H, Yin C and Xu D: The correlation between glucose fluctuation from self-monitored blood glucose and the major adverse cardiac events in diabetic patients with acute coronary syndrome during a 6-month follow-up by WeChat application. Clin Chem Lab Med. 56:2119–2124. 2018.PubMed/NCBI View Article : Google Scholar
39	Glucose tolerance and mortality. Comparison of WHO and American Diabetes Association diagnostic criteria. The DECODE study group. European Diabetes Epidemiology Group. Diabetes Epidemiology: Collaborative analysis Of Diagnostic criteria in Europe. Lancet. 354:617–621. 1999.PubMed/NCBI
40	Curtin JF, Donovan M and Cotter TG: Regulation and measurement of oxidative stress in apoptosis. J Immunol Methods. 265:49–72. 2002.PubMed/NCBI View Article : Google Scholar
41	Nikoletopoulou V, Markaki M, Palikaras K and Tavernarakis N: Crosstalk between apoptosis, necrosis and autophagy. Biochim Biophys Acta. 1833:3448–3459. 2013.PubMed/NCBI View Article : Google Scholar
42	Hussain T, Tan B, Yin Y, Blachier F, Tossou MC and Rahu N: Oxidative stress and inflammation: What Polyphenols Can Do for Us? Oxid Med Cell Longev. 2016(7432797)2016.PubMed/NCBI View Article : Google Scholar
43	Shen Y, Zhang Q, Huang Z, Zhu J, Qiu J, Ma W, Yang X, Ding F and Sun H: Isoquercitrin delays denervated soleus muscle atrophy by inhibiting oxidative stress and inflammation. Front Physiol. 11(988)2020.PubMed/NCBI View Article : Google Scholar
44	Meng XM, Ren GL, Gao L, Yang Q, Li HD, Wu WF, Huang C, Zhang L, Lv XW and Li J: NADPH oxidase 4 promotes cisplatin-induced acute kidney injury via ROS-mediated programmed cell death and inflammation. Lab Invest. 98:63–78. 2018.PubMed/NCBI View Article : Google Scholar
45	Sedeek M, Montezano AC, Hebert RL, Gray SP, Di Marco E, Jha JC, Cooper ME, Jandeleit-Dahm K, Schiffrin EL, Wilkinson-Berka JL and Touyz RM: Oxidative stress, Nox isoforms and complications of diabetes-potential targets for novel therapies. J Cardiovasc Transl Res. 5:509–518. 2012.PubMed/NCBI View Article : Google Scholar
46	Sedeek M, Nasrallah R, Touyz RM and Hebert RL: NADPH oxidases, reactive oxygen species, and the kidney: Friend and foe. J Am Soc Nephrol. 24:1512–1518. 2013.PubMed/NCBI View Article : Google Scholar
47	Chu FF, Esworthy RS, Shen B, Gao Q and Doroshow JH: Dexamethasone and Tofacitinib suppress NADPH oxidase expression and alleviate very-early-onset ileocolitis in mice deficient in GSH peroxidase 1 and 2. Life Sci. 239(116884)2019.PubMed/NCBI View Article : Google Scholar
48	Lindquist RL, Bayat-Sarmadi J, Leben R, Niesner R and Hauser AE: NAD(P)H oxidase activity in the small intestine is predominantly found in enterocytes, not professional phagocytes. Int J Mol Sci. 19(1365)2018.PubMed/NCBI View Article : Google Scholar
49	Yan J, Wang C, Jin Y, Meng Q, Liu Q, Liu Z, Liu K and Sun H: Catalpol ameliorates hepatic insulin resistance in type 2 diabetes through acting on AMPK/NOX4/PI3K/AKT pathway. Pharmacol Res. 130:466–480. 2018.PubMed/NCBI View Article : Google Scholar
50	Sun L, Li W, Li W, Xiong L, Li G and Ma R: Astragaloside IV prevents damage to human mesangial cells through the inhibition of the NADPH oxidase/ROS/Akt/NF-κB pathway under high glucose conditions. Int J Mol Med. 34:167–176. 2014.PubMed/NCBI View Article : Google Scholar
51	Chen F, Qian LH, Deng B, Liu ZM, Zhao Y and Le YY: Resveratrol protects vascular endothelial cells from high glucose-induced apoptosis through inhibition of NADPH oxidase activation-driven oxidative stress. CNS Neurosci Ther. 19:675–681. 2013.PubMed/NCBI View Article : Google Scholar
52	Chowdhury SR, Saleh A, Akude E, Smith DR, Morrow D, Tessler L, Calcutt NA and Fernyhough P: Ciliary neurotrophic factor reverses aberrant mitochondrial bioenergetics through the JAK/STAT pathway in cultured sensory neurons derived from streptozotocin-induced diabetic rodents. Cell Mol Neurobiol. 34:643–649. 2014.PubMed/NCBI View Article : Google Scholar
53	Li Q, Lin Y, Wang S, Zhang L and Guo L: GLP-1 inhibits high-glucose-induced oxidative injury of vascular endothelial cells. Sci Rep. 7(8008)2017.PubMed/NCBI View Article : Google Scholar
54	Liu M, Yan L, Liang B, Li Z, Jiang Z, Chu C and Yang J: Hydrogen sulfide attenuates myocardial fibrosis in diabetic rats through the JAK/STAT signaling pathway. Int J Mol Med. 41:1867–1876. 2018.PubMed/NCBI View Article : Google Scholar
55	Marrero MB, Banes-Berceli AK, Stern DM and Eaton DC: Role of the JAK/STAT signaling pathway in diabetic nephropathy. Am J Physiol Renal Physiol. 290:F762–F768. 2006.PubMed/NCBI View Article : Google Scholar
56	Wang X, Shaw S, Amiri F, Eaton DC and Marrero MB: Inhibition of the Jak/STAT signaling pathway prevents the high glucose-induced increase in tgf-beta and fibronectin synthesis in mesangial cells. Diabetes. 51:3505–3509. 2002.PubMed/NCBI View Article : Google Scholar
57	Yoshikawa H, Matsubara K, Qian GS, Jackson P, Groopman JD, Manning JE, Harris CC and Herman JG: SOCS-1, a negative regulator of the JAK/STAT pathway, is silenced by methylation in human hepatocellular carcinoma and shows growth-suppression activity. Nat Genet. 28:29–35. 2001.PubMed/NCBI View Article : Google Scholar
58	Das A, Salloum FN, Durrant D, Ockaili R and Kukreja RC: Rapamycin protects against myocardial ischemia-reperfusion injury through JAK2-STAT3 signaling pathway. J Mol Cell Cardiol. 53:858–869. 2012.PubMed/NCBI View Article : Google Scholar
59	Sun X, Chen RC, Yang ZH, Sun GB, Wang M, Ma XJ, Yang LJ and Sun XB: Taxifolin prevents diabetic cardiomyopathy in vivo and in vitro by inhibition of oxidative stress and cell apoptosis. Food Chem Toxicol. 63:221–232. 2014.PubMed/NCBI View Article : Google Scholar