Cytochrome c modulates the mitochondrial signaling pathway and polymorphonuclear neutrophil apoptosis in bile duct-ligated rats

Deng,Xuesong; Deng,Tongming; Ni,Yong; Zhan,Yongqiang; Huang,Wenlong; Liu,Jianfeng; Liao,Caixian

doi:10.3892/etm.2016.3313

Cytochrome c modulates the mitochondrial signaling pathway and polymorphonuclear neutrophil apoptosis in bile duct-ligated rats

Authors:
- Xuesong Deng
- Tongming Deng
- Yong Ni
- Yongqiang Zhan
- Wenlong Huang
- Jianfeng Liu
- Caixian Liao
View Affiliations

Affiliations: Department of Hepatobiliary Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Department of General Surgery, Baoan Central Hospital of Shenzhen, Shenzhen, Guangdong 518102, P.R. China, Department of Hepatobiliary Surgery, The Second People's Hospital of Shenzhen (The First Affiliated Hospital of Shenzhen University), Shenzhen, Guangdong 518035, P.R. China
Published online on: May 9, 2016 https://doi.org/10.3892/etm.2016.3313
Pages: 333-342

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

It has been observed that polymorphonuclear neutrophils (PMN) increase in number and function during obstructive jaundice (OJ). However, the precise mechanisms underlying PMN apoptosis during OJ remain poorly understood. The aim of the present study was to investigate the modulation of cytochrome c (Cytc) on the mitochondrial signaling pathway in bile duct‑ligated (BDL) rats and the effect on PMN apoptosis following the intravenous administration of Cytc. Rats were randomly divided into four groups: A control group, a sham group, a BDL group and a BDL + Cytc group (rats with common bile duct ligation as well as Cytc intravenous injection). Blood samples were collected from the inferior vein cava for biochemical analysis and separation of the PMN. PMN apoptosis was evaluated using flow cytometry. The mitochondrial membrane potential (ΔΨm) of PMN was detected by rhodamine‑123 staining. The Cytc protein expression levels were examined using western blotting. PMN mitochondria were observed using transmission electron microscopy. The results of the present study revealed that the PMN apoptosis rate in rats decreased gradually from 12 to 72 h following BDL to levels that were significantly lower than those of the control group and the sham group. Compared with the corresponding time point of the BDL group, the BDL + Cytc group showed a significantly increased PMN apoptosis rate. The mean fluorescence intensity (MFI) of ΔΨm decreased from 12 to 72 h following BDL, and was significantly increased compared with the control and sham groups. MFI in the BDL + Cytc group was higher compared with that in the BDL group. Cytc expression levels increased in the mitochondria and decreased in the cytoplasm from the 12 to 72 h in the BDL group, which was significantly different from that in the control and sham groups at the corresponding time points. Compared with the BDL group, Cytc expression levels in the cytoplasm for the BDL + Cytc group tended to gradually and significantly increase. Morphological changes in PMN mitochondria were marginal in BDL rats and marked in the BDL + Cytc group. In the BDL rats, PMN apoptosis was inhibited, a process induced by the mitochondrial apoptotic signaling pathway in which Cytc has an important role. High ΔΨm in the mitochondria and decreased Cytc expression levels in the cytoplasm result in PMN apoptosis inhibition. Intravenous injection of Cytc may help compensate for the lack of Cytc proteins in the cytoplasm, inducing PMN apoptosis following BDL.

View Figures

View References

1	Smith JA: Neutrophils, host defense, and inflammation: A double-edged sword. J Leuko Biol. 56:672–686. 1994.PubMed/NCBI
2	Tsuji K, Kubota Y, Yamamoto S, Yanagitani K, Amoh Y, Takaoka M, Ogura M, Kin H and Inoue K: Increased neutrophil chemotaxis in obstructive jaundice: An in vitro experiment in rats. J Gastroenterol Hepatol. 14:457–463. 1999. View Article : Google Scholar : PubMed/NCBI
3	Takaoka M, Kubota Y, Tsuji K, Yamamoto S, Ogura M, Yanagitani K, Shimatani M, Shibatani N and Inoue K: Human neutrophil functions in obstructive jaundice. Hepatogastroenterology. 48:71–75. 2001.PubMed/NCBI
4	Shibatani N, Yamamoto S, Kubota Y, Tsuji K, Takaoka M, Amoh Y, Matsushita M, Shimatani M, Imai Y and Inoue K: Neutrophil chemotaxis in bile duct-obstructed rats, and effect of internal biliary drainage. Hepatogastroenterology. 49:918–923. 2002.PubMed/NCBI
5	Shimatani M, Tsuji K, Aze Y, Yamamoto S, Shibatani N, Imai Y, Takamido S, Kubota Y and Okazaki K: Effects of obstructive jaundice on neutrophil production and acquisition of chemotactic activity in the bone marrow. J Gastroenterol Hepatol. 20:117–125. 2005. View Article : Google Scholar : PubMed/NCBI
6	Badger SA, Jones C, McCaigue M, Clements BW, Parks RW, Diamond T, McCallion K and Taylor MA: Cytokine response to portal endotoxaemia and neutrophil stimulation in obstructive jaundice. Eur J Gastroenterol Hepatol. 24:25–32. 2012. View Article : Google Scholar : PubMed/NCBI
7	Chitnis D, Dickerson C, Munster AM and Winchurch RA: Inhibition of apoptosis in polymorphonuclear neutrophils from burn patients. J Leuko Biol. 59:835–839. 1996.PubMed/NCBI
8	Ertel W, Keel M, Infanger M, Ungethum U, Steckholzer U and Trentz O: Circulating mediators in serum of injured patients with septic complications inhibit neutrophil apoptosis through up-regulation of protein-tyrosine phosphorylation. J Trauma. 44:767–775; discussion 775–776. 1998. View Article : Google Scholar : PubMed/NCBI
9	Fanning NF, Kell MR, Shorten GD, Kirwan WO, Bouchier-Hayes D, Cotter TG and Redmond HP: Circulating granulocyte macrophage colony-stimulating factor in plasma of patients with the systemic inflammatory response syndrome delays neutrophil apoptosis through inhibition of spontaneous reactive oxygen species generation. Shock. 11:167–174. 1999. View Article : Google Scholar : PubMed/NCBI
10	Chiu DF, Chen JC, Chen HM, Ng CJ, Shyr MH and Chen MF: Results of treating severe acute pancreatitis with gabexate is associated with neutrophil apoptosis activity. Hepatogastroenterology. 50:553–558. 2003.PubMed/NCBI
11	Matsuda T, Saito H, Fukatsu K, Han I, Inoue T, Furukawa S, Ikeda S and Hidemura A: Cytokine-modulated inhibition of neutrophil apoptosis at local site augments exudative neutrophil functions and reflects inflammatory response after surgery. Surgery. 129:76–85. 2001. View Article : Google Scholar : PubMed/NCBI
12	Simon HU: Neutrophil apoptosis pathways and their modifications in inflammation. Immunol Rev. 193:101–110. 2003. View Article : Google Scholar : PubMed/NCBI
13	Maier RV: Pathogenesis of multiple organ dysfunction syndrome-endotoxin, inflammatory cells, and their mediators: Cytokines and reactive oxygen species. Surg Infect (Larchmt). 1:197–204; discussion 204–205. 2000. View Article : Google Scholar : PubMed/NCBI
14	Colotta F, Re F, Polentarutti N, Sozzani S and Mantovani A: Modulation of granulocyte survival and programmed cell death by cytokines and bacterial products. Blood. 80:2012–2020. 1992.PubMed/NCBI
15	Esmann L, Idel C, Sarkar A, Hellberg L, Behnen M, Möller S, van Zandbergen G, Klinger M, Köhl J, Bussmeyer U, et al: Phagocytosis of apoptotic cells by neutrophil granulocytes: Diminished proinflammatory neutrophil functions in the presence of apoptotic cells. J Immunol. 184:391–400. 2010. View Article : Google Scholar : PubMed/NCBI
16	Twomey C and McCarthy JV: Pathways of apoptosis and importance in development. J Cell Mol Med. 9:345–359. 2005. View Article : Google Scholar : PubMed/NCBI
17	Schwartz JT, Barker JH, Kaufman J, Fayram DC, McCracken JM and Allen LA: Francisella tularensis inhibits the intrinsic and extrinsic pathways to delay constitutive apoptosis and prolong human neutrophil lifespan. J Immunol. 188:3351–3363. 2012. View Article : Google Scholar : PubMed/NCBI
18	Green DR: Apoptotic pathways: Ten minutes to dead. Cell. 121:671–674. 2005. View Article : Google Scholar : PubMed/NCBI
19	Ottonello L, Frumento G, Arduino N, Bertolotto M, Dapino P, Mancini M and Dallegri F: Differential regulation of spontaneous and immune complex-induced neutrophil apoptosis by proinflammatory cytokines. Role of oxidants, Bax and caspase-3. J Leukoc Biol. 72:125–132. 2002.PubMed/NCBI
20	Le'Negrate G, Rostagno P, Auberger P, Rossi B and Hofman P: Downregulation of caspases and Fas ligand expression, and increased lifespan of neutrophils after transmigration across intestinal epithelium. Cell Death Differ. 10:153–162. 2003. View Article : Google Scholar : PubMed/NCBI
21	Nauseef WM: Isolation of human neutrophils from venous blood. Methods Mol Biol. 412:15–20. 2007. View Article : Google Scholar : PubMed/NCBI
22	Scorrano L: Opening the doors to cytochrome c: Changes in mitochondrial shape and apoptosis. Int J Biochem Cell Biol. 41:1875–1883. 2009. View Article : Google Scholar : PubMed/NCBI
23	Gustafsson AB and Gottlieb RA: Heart mitochondria: Gates of life and death. Cardiovasc Res. 77:334–343. 2008. View Article : Google Scholar : PubMed/NCBI
24	Yang Y, Xing D, Zhou F and Chen Q: Mitochondrial autophagy protects against heat shock-induced apoptosis through reducing cytosolic cytochrome c release and downstream caspase-3 activation. Biochem Biophys Res Commun. 395:190–195. 2010. View Article : Google Scholar : PubMed/NCBI
25	Mei Y, Yong J, Liu H, Shi Y, Meinkoth J, Dreyfuss G and Yang X: tRNA binds to cytochrome c and inhibits caspase activation. Mol Cell. 37:668–678. 2010. View Article : Google Scholar : PubMed/NCBI
26	Kurtoglu M and Lampidis TJ: From delocalized lipophilic cations to hypoxia: Blocking tumor cell mitochondrial function leads to therapeutic gain with glycolytic inhibitors. Mol Nutr Food Res. 53:68–75. 2009. View Article : Google Scholar : PubMed/NCBI
27	Fox S, Leitch AE, Duffin R, Haslett C and Rossi AG: Neutrophil apoptosis: Relevance to the innate immune response and inflammatory disease. J Innate Immun. 2:216–227. 2010. View Article : Google Scholar : PubMed/NCBI
28	Taneja R, Parodo J, Jia SH, Kapus A, Rotstein OD and Marshall JC: Delayed neutrophil apoptosis in sepsis is associated with maintenance of mitochondrial transmembrane potential and reduced caspase-9 activity. Crit Care Med. 32:1460–1499. 2004. View Article : Google Scholar : PubMed/NCBI
29	Kole AJ, Knight ER and Deshmukh M: Activation of apoptosis by cytoplasmic microinjection of cytochrome c. J Vis Exp pii. 27732011.
30	Kroemer G, Dallaporta B and Resche-Rigon M: The mitochondrial death/life regulator in apoptosis and necrosis. Annu Rev Physiol. 60:619–642. 1998. View Article : Google Scholar : PubMed/NCBI
31	Lundqvist-Gustafsson H, Norrman S, Nilsson J and Wilsson A: Involvement of p38-mitogen protein kinase in Staphylococcus aureus-induced neutrophil apoptosis. J Leukoc Biol. 70:642–648. 2001.PubMed/NCBI
32	Altavilla D, Saitta A, Squadrito G, Galeano M, Venuti SF, Guarini S, Bazzani C, Bertolini A, Caputi AP and Squadrito F: Evidence for a role of nuclear factor-kappaB in acute hypovolemic hemorrhagic shock. Surgery. 131:50–58. 2002. View Article : Google Scholar : PubMed/NCBI
33	Sousa LP, Lopes F, Silva DM, Tavares LP, Vieira AT, Rezende BM, Carmo AF, Russo RC, Garcia CC, Bonjardim CA, et al: PDE4 inhibition drives resolution of neutrophilic inflammation by inducing apoptosis in a PKA-PI3K/Akt-dependent and NF-kappaB-independent manner. J Leukoc Biol. 87:895–904. 2010. View Article : Google Scholar : PubMed/NCBI
34	Fernandes CA, Fievez L, Ucakar B, Neyrinck AM, Fillee C, Huaux F, Delzenne NM, Bureau F and Vanbever R: Nicotinamide enhances apoptosis of G(M)-CSF-treated neutrophils and attenuates endotoxin-induced airway inflammation in mice. Am J Physiol Lung Cell Mol Physiol. 300:L354–L361. 2011. View Article : Google Scholar : PubMed/NCBI
35	Paunel-Görgülü A, Zörnig M, Lögters T, Altrichter J, Rabenhorst U, Cinatl J, Windolf J and Scholz M: Mcl-1 mediated impairment of the intrinsic apoptosis pathway in circulating neutrophils from critically ill patients can be overcome by Fas stimulation. J Immunol. 183:6198–6206. 2009. View Article : Google Scholar : PubMed/NCBI
36	Nwakoby IE, Reddy K, Patel P, Shah N, Sharma S, Bhaskaran M, Gibbons N, Kapasi AA and Singhal PC: Fas-mediated apoptosis of neutrophils in sera of patients with infection. Infect Immun. 69:3343–3349. 2001. View Article : Google Scholar : PubMed/NCBI
37	Dunican AL, Lenemoth SJ, Grutkoski P, Ayala A and Simms HH: TNF alpha-induced suppression of PMN apoptosis is mediated through interleukin-8 production. Shock. 14:284–288; discussion 288–289. 2000. View Article : Google Scholar : PubMed/NCBI
38	Geering B and Simon HU: A novel signaling pathway in TNFα-induced neutrophil apoptosis. Cell Cycle. 10:2821–2822. 2011. View Article : Google Scholar : PubMed/NCBI
39	Maianski NA, Mul FP, van Buul JD, Roos D and Kuijpers TW: Granulocyte colony-stimulating factor inhibits the mitochondria-dependent activation of caspase-3 in neutrophils. Blood. 99:672–679. 2002. View Article : Google Scholar : PubMed/NCBI
40	Maianski NA, Roos D and Kuijpers TW: Tumor necrosis factor alpha induces a caspase-independent death pathway in human neutrophils. Blood. 101:1987–1995. 2003. View Article : Google Scholar : PubMed/NCBI
41	Maianski NA, Geissler J, Srinivasula SM, Alnemri ES, Roos D and Kuijpers TW: Functional characterization of mitochondria in neutrophils: A role restricted to apoptosis. Cell Death Differ. 11:143–153. 2004. View Article : Google Scholar : PubMed/NCBI