Neutrophil depletion reduces interstitial cajal-like cell injury and alleviates inflammation-induced motor dysfunction in guinea-pig gallbladder during acute cholecystitis

Document Type : Original Article


1 Department of Gastroenterology, The First Affiliated Hospital of Chongqing Medical University, No. 1, Youyi Road, Yuzhong District, Chongqing 400016, China

2 Department of Gastroenterology, Renmin Hospital of Wuhan University, No. 238, Jiefang Road, Wuhan 430060, Hubei Province, China

3 Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, No. 1, Youyi Road, Yuzhong District, Chongqing 400016, China


Objective(s): Gallbladder interstitial Cajal-like cells (ICLCs) are known as some of the players in the complex motility mechanisms affecting gallbladder motility. This study aims to explore the mechanism of guinea-pig gallbladder motility disorders during Acute Cholecystitis (AC), focusing on the relationships between neutrophil alterations, gallbladder ICLCs, and smooth muscle contractility.
Materials and Methods: Forty-eight guinea pigs were randomly divided into four groups: normal, sham, common bile duct ligation (CBDL), and anti-PMN (anti-polymorphonuclear antibody treated +CBDL). Hematoxylin and eosin-stained slides from each gallbladder sample were examined for inflammation, and myeloperoxidase (MPO) activity was evaluated. The contractile response of gallbladder muscle to Ach, CCK-8, and KCl was registered by a tension transducer, and ultrastructure features of ICLCs were observed.
Results: Pretreatment with anti-PMN significantly reduced the circulating neutrophils by 80% and also considerably decreased the gallbladder MPO activity by 52.9% compared with the CBDL group (P<0.05). After adding Ach, CCK-8, and KCl, the contraction ability in CBDL and anti-PMN groups was lower than those of normal and sham groups (P<0.05), and they were increased substantially in the anti-PMN group compared with the CBDL group (P<0.05). Transmission electron microscopy confirmed that the cytoplasm of the neutrophils was full of granules, and neutrophils contacted closely with ICLCs. The ultrastructure of ICLCs in the anti-PMN group was less inflamed and the endoplasmic reticulum was mildly dilated, and cell processes also increased.
Conclusion: Anti-PMN could relieve the ultrastructure injury of ICLCs and alleviate gallbladder dysmotility during AC. Neutrophils may damage gallbladder ICLCs at first followed by dysmotility.


1.    Treinen C, Lomelin D, Krause C, Goede M, Oleynikov D. Acute acalculous cholecystitis in the critically ill: Risk factors and surgical strategies. Langenbecks Arch Surg 2015; 400:421-427.
2.    Xiao ZL, Chen Q, Biancani P, Behar J. Abnormalities of gallbladder muscle associated with acute inflammation in guinea pigs. Am J Physiol Gastrointest Liver Physiol 2001; 281:G490- G497.
3.    Parkman HP, Bogar LJ, Bartula LL, Pagano AP, Thomas RM, Myers SI. Effect of experimental acalculous cholecystitis on gallbladder smooth muscle contractility. Dig Dis Sci 1999; 44:2235-2243.
4.    Gomez-Pinilla PJ, Morales S, Camello-Almaraz C, Moreno R, Pozo MJ, Camello PJ. Changes in guinea pig gallbladder smooth muscle Ca2+ homeostasis by acute acalculous cholecystitis. Am J Physiol Gastrointest Liver Physiol 2006; 290:G14-G22.
5.    Pasternak A, Szura M, Mazur M, Mróz I, Matyja M, Matyja A. Number and distribution of interstitial cells of Cajal in human gallbladder. Folia Med Cracov 2014; 54: 71-77.
6.    Huang ZP, Qiu H, Yu BP. Distribution changes of interstitial cells of Cajal during cholesterol gallstone formation in guinea pigs fed a high cholesterol diet. Int J Clin Exp Pathol 2018; 11:1653–1659.
7.    Yang XJ, Yang J, Liu Z, Yang G, Shen ZJ. Telocytes damage in endometriosis-affected rat oviduct and potential impact on fertility. J Cell Mol Med 2015; 19:452–462.
8.    Popescu LM, Gherghiceanu M, Cretoiu D, Radu E. The connective connection: interstitial cells of Cajal (ICC) and ICC-like cells establish synapses with immunoreactive cells. Electron microscope study in situ. J Cell Mol Med 2005; 9:714–730. 
9.    Bettolli M, De Carli C, Cornejo-Palma D, Jolin-Dahel K, Wang XY, Huizinga J, Krantis A, et al. Interstitial cell of Cajal loss correlates with the degree of inflammation in the human appendix and reverses after inflammation. J Pediatr Surg 2012; 47:1891-1899.
10.    Fan Y, Wu S, Fu B, Weng C, Wang X. The role of interstitial Cajal-like cells in the formation of cholesterol stones in guinea pig gallbladder. Hepatol Int 2015; 9:612–620.
11.    Pasternak A, Matyja A, Gil K, Gajda M, Tomaszewski KA, Gajda M, Tomaszewski KA, et al. Interstitial cajal-like cells and bile lithogenicity in the pathogenesis of gall-stone disease. Pol Przegl Chir 2013; 85:311-316..
12.    Huang ZP, Qiu H, Yang Y, Zhang L, Yang B, Lin MJ, Yu BP. The role of interstitial cells of cajal in acute cholecystitis in guinea pig gallbladder. Cell Physiol Biochem 2016; 38:1775-1784. 
13.    Huang ZP, Qiu H, Yang Y, Yu BP. Effect of Neutrophils on Gallbladder Interstitial Cajal-Like Cells in Guinea Pig Model of Acute Cholecystitis. Cell Physiol Biochem 2016; 39:2033–2043. 
14.    Huang ZP, Qiu H, Yu BP. Acute cholecystitis reduces interstitial cells of cajal in porcine gallbladder through decreased mRNA synthesis. Cell Physiol Biochem 2018; 47:535–544.
15.    Zhang L, Pan C, Yang B, Xiao Y, Yu B. Enhanced expression of cystathionine β-synthase and cystathionine γ-lyase during acute cholecystitis-induced gallbladder inflammation. PLoS One 2013; 8:e82711.
16.    Moxon-Emre I, Schlichter LC. Neutrophil depletion reduces blood-brain barrier breakdown, axon injury, and inflammation after intracerebral hemorrhage. J Neuropathol Exp Neurol 2011, 70:218-235.
17.    Wang AJ, Wang TE, Lin CC, Lin SC, Shih SC. Clinical predictors of severe gallbladder complications in acute acalculous cholecystitis. World J Gastroenterol 2003; 9:2821-2823.
18.    Soylu S, Aydin C, Bagcivan I, Yildirim S, Koyuncu A, Topcu O, Arici S. Effects of NO/L-arginine pathway on gallbladder contractility in bile duct ligated guinea pigs. J Surg Res 2009; 155:70-76.
19.    Panteghini M, Malchiodi A, Calarco M, Bonora R. Clinical and diagnostic significance of aspartate aminotransferase isoenzymes in sera of patients with liver diseases. J Clin Chem Clin Biochem 1984;22:153-158.
20.    Stein TA, Burns GP, Wise L. Diagnostic value of liver function tests in bile duct obstruction. J Surg Res 1989; 46:226-229.
21.    Bouchery T, Harris N. Neutrophil-macrophage cooperation and its impact on tissue repair. Immunol Cell Biol 2019; 97:289–298.
22.    Ryu JK, Tran KC, McLarnon JG. Depletion of neutrophils reduces neuronal degeneration and inflammatory responses induced by quinolinic acid in vivo. Glia 2007; 55:439-451.
23.    Lavoie B, Balemba OB, Nelson MT, Ward SM, Mawe GM. Morphological and physiological evidence for interstitial cell of Cajal-like cells in the guinea pig gallbladder. J Physiol 2007; 579:487-501.
24.    Lin MJ, Chen L, Huang ZP, Qiu H, Yu BP. Neutrophils injure gallbladder interstitial Cajal-like cells in a guinea pig model of acute cholecystitis. J Cell Physiol 2019; 234:4291-4301.
25.    Kaji N, Horiguchi K, Iino S, Nakayama S, Ohwada T, Otani Y, Firman, et al. Nitric oxide-induced oxidative stress impairs pacemaker function of murine interstitial cells of Cajal during inflammation. Pharmacol Res 2016; 111:838–848.
26.    Díaz-Flores L, Gutiérrez R, García MP, Sáez FJ, Aparicio F, Díaz-Flores L Jr, Madrid JF. Uptake and intracytoplasmic storage of pigmented particles by human CD34+ stromal cells/telocytes: endocytic property of telocytes. J Cell Mol Med 2014;18:2478-2487.
27.    Xu D, Yu BP, Luo HS, Chen LD. Control of gallbladder contractions by cholecystokinin through cholecystokinin-A receptors on gallbladder interstitial cells of Cajal. World J Gastroenterol 2008; 14:2882–2887.
28.    Parkman HP, Pagano AP, Ringold MA, Ryan JP. Effect of modulating voltage-dependent calcium channels on cholecystokinin and acetylcholine-induced contractions of the guinea pig gallbladder. Regul Pept 1996; 63:31-37.