Evaluation of intestinal injury, inflammatory response and oxidative stress following intracerebral hemorrhage in mice

Cheng,Yijun; Zan,Jieyu; Song,Yaying; Yang,Guoyuan; Shang,Hanbing; Zhao,Weiguo

doi:10.3892/ijmm.2018.3755

Evaluation of intestinal injury, inflammatory response and oxidative stress following intracerebral hemorrhage in mice

Authors:
- Yijun Cheng
- Jieyu Zan
- Yaying Song
- Guoyuan Yang
- Hanbing Shang
- Weiguo Zhao
View Affiliations

Affiliations: Department of Neurosurgery, Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, P.R. China, Department of Pediatrics, Nantong First People's Hospital, Nantong, Jiangsu 226001, P.R. China, Department of Neurology, Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, P.R. China, Department of Neurosurgery, Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, P.R. China
Published online on: July 4, 2018 https://doi.org/10.3892/ijmm.2018.3755
Pages: 2120-2128

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

Intestinal injury is a common complication following intracerebral hemorrhage (ICH), which leads to malnutrition, impaired immunity and unsatisfactory prognosis. Previous studies have revealed the pathogenesis of intestinal injury following traumatic brain injury using ischemic stroke models. However, the effects of ICH on intestinal injury remain unknown. The present study aimed to investigate the pathological alterations and molecular mechanism, as well as the time course of intestinal injury following ICH in mice. Male C57BL/6 mice were randomly divided into the following seven groups (n=6 mice/group): Control group, which underwent a sham operation, and six ICH groups (2, 6, 12 and 24 h, and days 3 and 7). The ICH model was induced by stereotactically injecting autologous blood in two stages into the brain. Subsequently, intestinal tissue was stained with hematoxylin and eosin for histopathological examination. Small intestinal motility was measured by charcoal meal test, and gut barrier dysfunction was evaluated by detecting the plasma levels of endotoxin. Quantitative polymerase chain reaction (qPCR), immunohistochemistry and ELISA analysis were performed to evaluate the mRNA and protein expression levels of inflammatory cytokines [interleukin (IL)‑1β, IL‑6, tumor necrosis factor‑α, intercellular adhesion molecule 1, monocyte chemotactic protein 1 and chemokine (C‑C motif) ligand‑5] in intestinal tissue and serum. Furthermore, intestinal leukocyte infiltration was detected by measuring myeloperoxidase activity. Oxidative stress was indirectly detected by measuring reactive oxygen species‑associated markers (malondialdehyde content and superoxide dismutase activity assays) and the mRNA and protein expression levels of antioxidant genes [nuclear factor (erythroid‑derived 2)‑like 2, manganese superoxide dismutase and heme oxygenase 1] by qPCR and western blot analysis. The results demonstrated that significant destruction of the gut mucosa, delayed small intestinal motility, intestinal barrier dysfunction, and increased inflammatory responses and oxidative stress occurred rapidly in response to ICH. These symptoms occurred as early as 2 h after ICH and persisted for 7 days. These findings suggested that ICH may induce immediate and persistent damage to gut structure and barrier function, which may be associated with upregulation of inflammation and oxidative stress markers.

View Figures

View References

1	Keep RF, Hua Y and Xi G: Intracerebral haemorrhage: Mechanisms of injury and therapeutic targets. Lancet Neurol. 11:720–731. 2012. View Article : Google Scholar : PubMed/NCBI
2	Adeoye O and Broderick JP: Advances in the management of intracerebral hemorrhage. Nat Rev Neurol. 6:593–601. 2010. View Article : Google Scholar : PubMed/NCBI
3	Lu WY, Rhoney DH, Boling WB, Johnson JD and Smith TC: A review of stress ulcer prophylaxis in the neurosurgical intensive care unit. Neurosurgery. 41:416–425; discussion 425–426. 1997. View Article : Google Scholar : PubMed/NCBI
4	Misra UK, Kalita J, Pandey S and Mandal SK: Predictors of gastrointestinal bleeding in acute intracerebral haemorrhage. J Neurol Sci. 208:25–29. 2003. View Article : Google Scholar : PubMed/NCBI
5	Yang TC, Li JG, Shi HM, Yu DM, Shan K, Li LX, Dong XY and Ren TH: Gastrointestinal bleeding after intracerebral hemorrhage: A retrospective review of 808 cases. Am J Med Sci. 346:279–282. 2013. View Article : Google Scholar
6	Shinohara Y, Yanagihara T, Abe K, Yoshimine T, Fujinaka T, Chuma T, Ochi F, Nagayama M, Ogawa A, Suzuki N, et al: III. Intracerebral hemorrhage. J Stroke Cerebrovasc Dis. 20(Suppl 4): S74–S99. 2011. View Article : Google Scholar : PubMed/NCBI
7	Wang WJ, Lu JJ, Wang YJ, Wang CX, Wang YL, Hoff K, Yang ZH, Liu LP, Wang AX and Zhao XQ; China National Stroke Registry (CNSR): Clinical characteristics, management, and functional outcomes in Chinese patients within the first year after intracerebral hemorrhage: Analysis from China National Stroke Registry. CNS Neurosci Ther. 18:773–780. 2012. View Article : Google Scholar : PubMed/NCBI
8	Li X, Hammer AM, Rendon JL and Choudhry MA: Intestine immune homeostasis after alcohol and burn injury. Shock. 43:540–548. 2015. View Article : Google Scholar : PubMed/NCBI
9	Phillips NA, Welc SS, Wallet SM, King MA and Clanton TL: Protection of intestinal injury during heat stroke in mice by interleukin-6 pretreatment. J Physiol. 593:739–753. 2015. View Article : Google Scholar :
10	Timmermans K, Sir Ö, Kox M, Vaneker M, de Jong C, Gerretsen J, Edwards M, Scheffer GJ and Pickkers P: Circulating iFABP Levels as a marker of intestinal damage in trauma patients. Shock. 43:117–120. 2015. View Article : Google Scholar
11	Hang CH, Shi JX, Li JS, Wu W and Yin HX: Alterations of intestinal mucosa structure and barrier function following traumatic brain injury in rats. World J Gastroenterol. 9:2776–2781. 2003. View Article : Google Scholar : PubMed/NCBI
12	Xu X, Zhu Y and Chuai J: Changes in serum ghrelin and small intestinal motility in rats with ischemic stroke. Anat Rec (Hoboken). 295:307–312. 2012. View Article : Google Scholar
13	Low G, Jaremko JL and Lomas DJ: Extravascular complications following abdominal organ transplantation. Clin Radiol. 70:898–908. 2015. View Article : Google Scholar : PubMed/NCBI
14	Doig CJ, Sutherland LR, Sandham JD, Fick GH, Verhoef M and Meddings JB: Increased intestinal permeability is associated with the development of multiple organ dysfunction syndrome in critically ill ICU patients. Am J Respir Crit Care Med. 158:444–451. 1998. View Article : Google Scholar : PubMed/NCBI
15	National Research Council: Guide for the care and use of laboratory animals. 8th edition. National Academies Press; Washington DC: pp. 1–246. 2011
16	Chiu CJ, McArdle AH, Brown R, Scott HJ and Gurd FN: Intestinal mucosal lesion in low-flow states. I. A morphological, hemodynamic, and metabolic reappraisal. Arch Surg. 101:478–483. 1970. View Article : Google Scholar : PubMed/NCBI
17	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
18	Molmenti EP, Ziambaras T and Perlmutter DH: Evidence for an acute phase response in human intestinal epithelial cells. J Biol Chem. 268:14116–14124. 1993.PubMed/NCBI
19	Liu XH, Yang YW, Dai HT, Cai SW, Chen RH and Ye ZQ: Protective role of adiponectin in a rat model of intestinal ischemia reperfusion injury. World J Gastroenterol. 21:13250–13258. 2015. View Article : Google Scholar : PubMed/NCBI
20	Arndt H, Kubes P, Grisham MB, Gonzalez E and Granger DN: Granulocyte turnover in the feline intestine. Inflammation. 16:549–559. 1992. View Article : Google Scholar : PubMed/NCBI
21	Nussbaum C, Klinke A, Adam M, Baldus S and Sperandio M: Myeloperoxidase: A leukocyte-derived protagonist of inflammation and cardiovascular disease. Antioxid Redox Signal. 18:692–713. 2013. View Article : Google Scholar
22	Xue B, Kasparek MS, Müller MH and Kreis ME: Modulation of intestinal afferent nerve sensitivity to inflammatory mediators following systemic endotoxin in mice. Neurogastroenterol Motil. 27:550–558. 2015. View Article : Google Scholar : PubMed/NCBI
23	Bohatschek M, Werner A and Raivich G: Systemic LPS injection leads to granulocyte influx into normal and injured brain: Effects of ICAM-1 deficiency. Exp Neurol. 172:137–152. 2001. View Article : Google Scholar : PubMed/NCBI
24	Hang CH, Shi JX, Li JS, Li WQ and Wu W: Expressions of intestinal NF-kappaB, TNF-α, and IL-6 following traumatic brain injury in rats. J Surg Res. 123:188–193. 2005. View Article : Google Scholar : PubMed/NCBI
25	Hang CH, Shi JX, Li JS, Li WQ and Yin HX: Upregulation of intestinal nuclear factor kappa B and intercellular adhesion molecule-1 following traumatic brain injury in rats. World J Gastroenterol. 11:1149–1154. 2005. View Article : Google Scholar : PubMed/NCBI
26	Hang CH, Shi JX, Tian J, Li JS, Wu W and Yin HX: Effect of systemic LPS injection on cortical NF-kappaB activity and inflammatory response following traumatic brain injury in rats. Brain Res. 1026:23–32. 2004. View Article : Google Scholar : PubMed/NCBI
27	Kalff JC, Schraut WH, Billiar TR, Simmons RL and Bauer AJ: Role of inducible nitric oxide synthase in postoperative intestinal smooth muscle dysfunction in rodents. Gastroenterology. 118:316–327. 2000. View Article : Google Scholar : PubMed/NCBI
28	Wehner S, Vilz TO, Stoffels B and Kalff JC: Immune mediators of postoperative ileus. Langenbecks Arch Surg. 397:591–601. 2012. View Article : Google Scholar : PubMed/NCBI
29	The FO, de Jonge WJ, Bennink RJ, van den Wijngaard RM and Boeckxstaens GE: The ICAM-1 antisense oligonucleotide ISIS-3082 prevents the development of postoperative ileus in mice. Br J Pharmacol. 146:252–258. 2005. View Article : Google Scholar : PubMed/NCBI
30	Wehner S, Schwarz NT, Hundsdoerfer R, Hierholzer C, Tweardy DJ, Billiar TR, Bauer AJ and Kalff JC: Induction of IL-6 within the rodent intestinal muscularis after intestinal surgical stress. Surgery. 137:436–446. 2005. View Article : Google Scholar : PubMed/NCBI
31	Marchiando AM, Shen L, Graham WV, Weber CR, Schwarz BT, Austin JR II, Raleigh DR, Guan Y, Watson AJ, Montrose MH, et al: Caveolin-1-dependent occludin endocytosis is required for TNF-induced tight junction regulation in vivo. J Cell Biol. 189:111–126. 2010. View Article : Google Scholar : PubMed/NCBI
32	Turner JR: ‘Putting the squeeze’ on the tight junction: Understanding cytoskeletal regulation. Semin Cell Dev Biol. 11:301–308. 2000. View Article : Google Scholar : PubMed/NCBI
33	Giudice A, Arra C and Turco MC: Review of molecular mechanisms involved in the activation of the Nrf2-ARE signaling pathway by chemopreventive agents. Methods Mol Biol. 647:37–74. 2010. View Article : Google Scholar : PubMed/NCBI
34	Nguyen T, Nioi P and Pickett CB: The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J Biol Chem. 284:13291–13295. 2009. View Article : Google Scholar : PubMed/NCBI
35	Jin W, Wang H, Ji Y, Hu Q, Yan W, Chen G and Yin H: Increased intestinal inflammatory response and gut barrier dysfunction in Nrf2-deficient mice after traumatic brain injury. Cytokine. 44:135–140. 2008. View Article : Google Scholar : PubMed/NCBI
36	Jin W, Wang HD, Hu ZG, Yan W, Chen G and Yin HX and Yin HX: Transcription factor Nrf2 plays a pivotal role in protection against traumatic brain injury-induced acute intestinal mucosal injury in mice. J Surg Res. 157:251–260. 2009. View Article : Google Scholar : PubMed/NCBI
37	Rangasamy T, Guo J, Mitzner WA, Roman J, Singh A, Fryer AD, Yamamoto M, Kensler TW, Tuder RM, Georas SN, et al: Disruption of Nrf2 enhances susceptibility to severe airway inflammation and asthma in mice. J Exp Med. 202:47–59. 2005. View Article : Google Scholar : PubMed/NCBI
38	Lee JM and Johnson JA: An important role of Nrf2-ARE pathway in the cellular defense mechanism. J Biochem Mol Biol. 37:139–143. 2004.PubMed/NCBI
39	Shang H, Yang D, Zhang W, Li T, Ren X, Wang X and Zhao W: Time course of Keap1-Nrf2 pathway expression after experimental intracerebral haemorrhage: Correlation with brain oedema and neurological deficit. Free Radic Res. 47:368–375. 2013. View Article : Google Scholar : PubMed/NCBI