Investigation of Changes in Liver Microanatomy in the Steatosis Model Created by Permanent Canula in Rats

Year 2022, Volume: 12 Issue: 3, 659 - 664, 28.09.2022

Asrın Nalbant , Bayram Ufuk Şakul , Ferruh Yücel

https://doi.org/10.33808/clinexphealthsci.948391

Abstract

Objective: The knowledge of nonalcoholic fatty liver disease (NAFLD) and Nonalcoholic Steatohepatitis (NASH) is limited to the findings from available suitable models for this disease. A number of rodent models have been described in which relevant liver pathology develops in an appropriate metabolic context. In this experimental study, it was aimed to create a new liver fat model by giving fat from the portal vein of rats and to visualize the changes in the liver with advanced microscopic techniques.
Methods: 28 female rats were used in the study. Permanent intraabdominal cannulas were inserted into the portal vein of the rats. Rats were randomly divided four group. Intralipid 20% substance was injected through cannula to the experimental groups during the test period. Control group received saline at the same rate. At the end of the experiment, the animals were visualized with a laser speckle microscope and livers were divided into sections according to the stereological method. The sections were painted with Hematoxylin-Eosin, Oil red o, Masson trichoma, Bodipy, Nile red. Sections were evaluated under a microscope.
Results: Ballooning, inflammation and fibrosis were observed in the 2 week intralipid group. In the 1 week intralipid group, the rate of parenchyma decreased while the sinusoid rate increased, and sinusoid rate increased significantly in the 2 week intralipid (p˂0.05).
Conclusion: According to the findings, steatohepatitis was detected in the 2 week intralipid, whereas only steatosis was observed in the 1 week intralipid. Thus, it was concluded that the newly formed rat model causes steatosis.

Keywords

steatosis, NAFLD, laser speckle, portal vein, confocal microscope

References

• Michael C. Kew. Obesity as a cause of hepatocellular carcinoma. Ann Hepatol. 2015; 14 (3):299-303.
• Ooi GJ, PaBurton PR, Doyle L, Wentworth JM, Bhathal PS, Sikaris K, Cowley MA, Roberts SK, Kemp W, O’Brien PE, Brown WA. Modified thresholds for fibrosis risk scores in nonalcoholic fatty liver disease are necessary in the obese. Obes Surg. 2017;27: 115-125.
• Takahashi Y, Fukusato T. Histopathology of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis. World J Gastroenterol. 2014; 20(42): 15539–15548.
• Takahashi Y, Soejima Y, Fukusato T. Animal models of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis. World J Gastroenterol. 2012; 18(19): 2300–2308.
• Botham KM, Napolitano M. Bravo E. The emerging role of disturbed CoQ metabolism in nonalcoholic fatty liver disease development and progression. Nutrients 2015;7(12):9834-9846.
• Schattenberg JM, Galle PR. Animal models of non-alcoholic steatohepatitis of mice and man. Dig Dis. 2010; 28 (1): 247-254.
• Hebbard L, George J. Animal models of nonalcoholic fatty liver disease Nat. Rev. Gastroenterol. Hepatol. 2011; 8:35.
• Ritsma L, Steller EJA, Ellenbroek, SIJ, Kranenburg O, Borel Rinkes IHNM, Rheenen JV. Surgical implantation of an abdominal imaging window for intravital microscopy. Nat Protoc. 2013; 8 (3): 583-594.
• Gervaz P, Scholl B, Gillet M. Permanent access tothe portal vein in rats: experimental model. European Surgical Research 2000; 32: 203-206.
• Meisel JA, Le HD, Meijer VE, Nose V, Gura KM, Mulkern RV, Sharif MRA, Puder M. Comparison of 5 intravenous lipid emulsions and their effects on hepatic steatosis in a murine model. Journal of Pediatric Surgery 2011; 46: 666-673.
• Kalish BT, Le HD, Gura KM, Bistrian BR, Puder M. A metabolic analysis of two intravenous lipid emulsions in an murine model. Plos one 2013; 8(4): e59653.
• Altunkaynak BZ, Ozbek E. Overweight and structural alterations of the liver in female rats fed a high-fat diet: A stereological and histological study. Turk J Gastroenterol. 2009; 20(2): 93-103.
• Bedi KS, Thomas YM, Davies CA, Dobbing J. Synapse-to-neuron ratios of the frontal and cerebellar cortex of 30-day-old and adult rats undernourished during early postnatal life. J Comp Neurol. 1980; 193:49–56.
• Imajo K, Yoneda M, Kessoku T, Ogawa Y, Maeda S, Sumida Y, Hyogo H, Eguchi Y, Wada K, Nakajima A. Rodent models of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis. Int J Mol Sci. 2013; 14 (11); 21833.
• Brunt EM. Nonalcoholic fatty liver disease: what the pathologist can tell the clinician. Dig Dis. 2012; 1: 61-68.
• Shih TH, Huang CE, Lee YE, Chen CL, Wang CH, Huang CJ, Cheng KW, Wu SC, Juang SE, Jawan B, Yang SC. Preoperative portal vein velocity/size and effect on living donor liver transplantation recipient hemodynamics during inferior vena cava clamping. Transplant Proc. 2016; 48(4): 1049-1051.
• Noorafshan A, Esmail-Zadeh B, Bahmanpour S, Poost-Pasand A. Early stereological changes in liver of Sprague-Dawley rats after streptozotocin injection. Indian J Gastroenterol. 2005; 24: 104-107.
• Yahyazedeh A, Altunkaynak BZ, Akgül N, Akgül HM. A histopathological and stereological study of liver damage in female rats caused by mercury vapor. Biotech Histochem. 2017; 92(5): 338-346.
• Brancatelli G, Furlan A, Calandra A, Dioguardi Burgio M. Hepatic sinusoidal dilatation. Abdom Radiol (NY). 2018 doi: 10.1007/s00261.018.1465-1468.
• Sakamoto M, Tsujikawa H, Effendi K, Ojima H, Harada K, Zen Y, Kondo F, Nakano M, Kage M, Sumida Y, Hashimoto E, Yamada G, Okanoue T, Koike K. Pathological findings of nonalcoholic steatohepatitis and nonalcoholic fatty liver disease. Pathol Int. 2017; 67 (1): 1-7.
• Fan JG, Qiao L. Commonly used animal models of nonalcoholic steatohepatitis. Hepatobiliary Pancreat Dis Int. 2009; 8 (3): 233-240.
• Lu XY, Xi T, Lau WY, Dong H, Xian ZH, Hua Yu, Zhu Z, Shen F, Wu MC, Cong WM. Pathobiological features of small hepatocellular carcinoma: Correlation between tumor size and biological behavior. J Cancer Res Clin Oncol. 2011;137 (4): 567-575.
• Leung C, Herath CB, Jia Z, Goodwin M, Mak KY, Watt MJ. Dietary glycotoxins exacerbate progression of experimental fatty liver disease. J Hepatol. 2014; 60: 832-838
• Choi YJ, Lee CH, Lee KY, Jung SH, Lee BH. Increased hepatic fatty acid uptake and esterification contribute to tetracyclineinduced steatosis in mice. Toxicol Sci.2015; 145(2): 273-282.
• Hirooka M, Koizumi Y, Miyake T, Ochi H, Tokumoto Y, Tada F, Matsuura B, Abe M, Hiasa Y. Nonalcoholic fatty liver disease: portal hypertension due to outflow block in patients without cirrhosis. Radiology 2015; 274(2): 597-604.
• Seifalian AM, Piasecki C, Agarwal A, Davidson BR. The effect of graded steatosis on flow in the hepatic parenchymal microcirculation. Transplantation 1999; 68(6): 780-784.
• Pereira ENGDS, Silvares RR, Flores EEI, Rodrigues KL, Ramos IP, Silva IJ, Machado MP, Miranda RA, Pazos-Moura CC, Gonçalves-de-Albuquerque CF, Faria-Neto HCC, Tibiriça E, Daliry A. Hepatic microvascular dysfunction and increased advanced glycation end products are components of nonalcoholic fatty liver disease. PLoS One 2017;12(6): e0179654.
• McCuskey RS, Ito Y, Robertson GR, McCuskey MK, Perry M, Farrell GC. Hepatic microvascular dysfunction during evolution of dietary steatohepatitis in mice. Hepatology 2004;40(2):386-393.
• Oh MK, Winn J, Poordad F. Review article: diagnosis and treatment of non-alcoholic fatty liver disease. Aliment Pharmacol Ther. 2008; 28: 503–522.
• Çolak Y, Tuncer I. Nonalcoholic liver diseases and steatohepatitis. J Ist Faculty Med 2010; 73:3.