Pretreatment with Recombinant Human Interleukin 2 (rhIL-2) Up-regulates PCNA-positive Cells after Partial Hepatectomy in Rat Liver

Jeon, Miae,Kwon, Hyun Jung,Kim, Yong Hyun,Han, Kook-Il,Nam, Kung-Woo,Baik, Yeongjun,Lee, Sunghee,Kim, Wan-Jong,Han, Man-Deuk 한국생물공학회 2014 Biotechnology and Bioprocess Engineering Vol.19 No.1

We prepared recombinant human interleukin-2 (rhIL-2) and studied its pretreated influence on liver regeneration and the blood profile in partially (67%) hepatectomized (PH) male Sprague-Dawley rats. Rats were injected in the tail vein with rhIL-2 three times per day for 3 consecutive days and 67% underwent a partial hepatectomy (PH). Five days after the PH, liver tissue and blood samples were analyzed for liver regeneration and hematological changes. The weight of the liver in the rhIL-2 pretreated groups increased in a dose-dependent manner; with the highest treatment ($24{\times}10^4$ IU/kg), the maximum liver weight of 88.6% was exhibited. The control group showed a gradual increase to 76.3% of the original liver weight. A histological analysis of the liver showed an increase in proliferating cell nuclear antigen (PCNA)-positive cells in rhIL-2 pretreated rat livers. The rate of hepatocyte proliferation also increased significantly in primary cultured rat liver cells following rhIL-2 treatment. These results suggest that pretreatment with rhIL-2 may play adjuvant roles in liver regeneration after PH.

Auxiliary partial orthotopic liver transplantation for adult onset type II citrullinemia

Bum Soo Kim,Sun Hyung Joo,Suk Hwan Lee,Jung Il Lee,Hyun Cheol Kim,남덕호,Ho Chul Park 대한외과학회 2011 Annals of Surgical Treatment and Research(ASRT) Vol.80 No.6

Adult-onset type II citrullinemia (CTLN2) is a disorder caused by an inborn error of metabolism affecting the liver. CTLN2 is an autosomal recessive disorder characterized by recurrent encephalopathy with hyperammonemia due to highly elevated plasma levels of citrulline and ammonia, caused by a deficiency of argininosuccinate synthetase in the liver. A small number of patients have undergone liver transplantation with favorable results. In Korea, the limitations of the deceased donor pool have made living donor liver transplantation a common alternative treatment option. We report the case of a patient with type II citrullinemia who was treated successfully with auxiliary partial orthotopic liver transplantation (APOLT) from a living donor. This is the first description of an APOLT for a patient with adult onset type II citrullinemia in Korea.

Catechol-O-Methyltransferase Activity from Regenerating Liver after Partial Hepatectomy in Rats

곽춘식,--,-- 대한의생명과학회 2005 Biomedical Science Letters Vol.11 No.1

The change of catechol-O-methyltransferase (COMT) activity during regeneration of rat liver was studied. Cytosolic, mitochondrial and microsomal COMTs activities were estimated in regenerating rat livers over a period of ten days after 70% (median and left lateral lobes) partial hepatectomy. The values of Km and Vmax in the hepatic enzymes were also measured. The activities of cytosolic and microsomal COMTs in regenerating rat liver after partial hepatectomy were found to be significantly increased between the second and the third day. Whereas the mitochondrial COMT activity did not change. The Vmax values of the cytosolic and microsomal COMTs in the regenerating rat liver were significantly increased at the second day after partial hepatectomy, however, the Km values of the above hepatic enzymes did not vary in all the experimental groups. Therefore, the results suggest that the biosynthesis of COMT was increased during the regeneration of rat liver.

쥐 재생간에서의 Arylamine N-Methyltransferase 활성도

주일,김여희,곽춘식 啓明大學校 醫科大學 2002 계명의대학술지 Vol.21 No.1

재생간에서 AMT 활성도 변동을 알아보기 위하여 쥐의 간엽을 부분 절제한 후 각 시기의 재생간에서 이 효소의 활성도를 측정하는 한편 혈청에서 이 효소의 활성도를 측정하였다. 또한 재생간에서 이 효소의 Km치와 Vmax치도 함께 측정하였다. 쥐 재생간의 AMT 활성도는 세포질과 미토콘드리아 분획에서는 간엽 절제 후 1일 및 2일에, 마이크로솜 분획에서는 간엽 절제 후 1일부터 3일까지 통계학적으로 유의한 증가를 나타내었다. 그러나 혈청에서 이 효소의 활성도는 변동이 없었다. 간엽 절제 후 2일의 쥐 재생간에서 세포질, 미토콘드리아 및 마이크로솜의 AMT의 Km치는 변동이 없었다. 그러나 이들 분획에서 이 효소의Vmax치는 유의한 증가를 나타내었다. 이상의 결과로 보아 쥐간의 AMT는 간재생이 활발한 시기의 재생간에서는 그 합성이 증가되는 효소로 생각된다. Changes of arylamine N-methyltransferase (AMT) activity during regeneration of rat liver was studied. Cytosolic, mitochondrial and microsomal AMT activities were determined in regenerating rat livers over a period of ten days after 70% (median and left lateral lobes) partial hepatectomy. The enzyme activity in serum, and Km and Vmax values of the hepatic enzyme were also measured. The activities of cytosolic and mitochondrial AMT were found to be significantly increased between the first and the second day of regenerating rat liver, whereas microsomal AMT increased between the first and the third day after partial hepatectpmy. On the other hand, the serum AMT activity did not change. The Vmax values of the cytosolic, mitochondrial and microsomal AMT in the regenerating rat liver were significantly increased at the second day after partial hepatectomy, however, the Km values of the above hepatic enzymes did not vary in all experimental groups. Therefore, the above results indicate that the biosynthesis of AMT was increased during the regeneration of rat liver.

MicroRNA-183 accelerates the proliferation of hepatocyte during liver regeneration through targeting programmed cell death protein 6

Hou Fangxing,Li Xing,Wang Yanfeng,Xiao Xiangzuo 한국유전학회 2022 Genes & Genomics Vol.44 No.8

Background: Liver regeneration is a highly orchestrated process concerning the modulation of various microRNAs (miRs). miR-183 was recently found to be involved in the process of liver regeneration, that miR-183 was remarkably up-regulated at 2-6 h after partial hepatectomy. Objective: This study was aimed to explore the mechanism of miR-183 in on liver regeneration. Methods: After partial hepatectomy (PH) or transfection, we measured the changes of miR-183 and programmed cell death protein 6 (PDCD6) levels in rats and the hepatocytes. The histopathology was observed with hematoxylin-eosin staining. The miR-183 mimic and inhibitor plasmids were intravenously injected into rats, and the liver weight/body weight ratio was calculated. The prediction of TargetScan and the validation of luciferase activity assay were employed to confirm the targeting relationship between miR-183 and PDCD6. The viability, apoptosis and cell cycle of transfected rat hepatocyte BRL-3A were determined via MTT and flow cytometry assays. Results: MiR-183 expression showed a contrary tendency with that of PDCD6 during liver regeneration. Enhanced miR-183 in rats could notably increase liver/body weight ratio, while its inhibition did conversely. Overexpressed PDCD6, a target of miR-183, repressed the viability and cell cycle in hepatocytes, whereas its silence led to contrary results. Overexpressed miR-183 in BRL-3A cells enhanced cell viability and promoted the cell cycle yet suppressed apoptosis, whereas its inhibition showed contrary results, which were offset by PDCD6.

Partial Hepatectomy in Acetylation-Deficient BubR1 Mice Corroborates that Chromosome Missegregation Initiates Tumorigenesis

Yoo-Kyung Lee,Inai Park,이현숙 대한내분비학회 2014 Endocrinology and metabolism Vol.29 No.4

Background: Aneuploidy has been suggested as one of the major causes of cancer from the time of Boveri. In support of this notion, many studies have shown that cancer cells exhibit aneuploidy. However, there are evidences that do not support the aneuploidy hypothesis. We have previously reported that the spindle assembly checkpoint protein BubR1 is acetylated in mitosis and that the acetylation of BubR1 is crucial for checkpoint maintenance and chromosome-spindle attachment. Mice heterozygous for acetylation-deficient BubR1 (K243R/+) spontaneously develop cancer with chromosome instability. As K243R/+ mice develop hepatocellular carcinoma, we set out to test if chromosome mis-segregation was the cause of their liver cancer. Methods: Primary hepatocytes in the regenerating liver after partial hepatectomy (PH) were analyzed and compared for various mitotic parameters. Results: Primary hepatocytes isolated from K243R/+ mice after PH displayed a marked increase of chromosome misalignment, accompanied by an increase of micronuclei. In comparison, the number of nuclei per cell and the centrosome numbers were not different between wild-type and K243R/+ mice. Taken together, chromosome mis-segregation provokes tumorigenesis in mouse liver. Conclusion: Our results corroborate that PH provides a reliable tool for assessing mitotic infidelity and cancer in mice.

Polydeoxyribonucleotide administration promotes the expression of proliferating cell nuclear antigen during liver regeneration in rats

윤광호,김완종,한상균,한국일,남궁우,한만덕 한국통합생물학회 2015 Animal cells and systems Vol.19 No.1

Polydeoxyribonucleotide (PDRN) is an agonist of the A2A adenosine receptors. We evaluated the effects of PDRN on liverregeneration induced by partial hepatectomy (PH) in rats, focusing on cell proliferation. Proliferating cell nuclear antigen(PCNA) as an indicator of cell proliferation was targeted to determine whether PDRN treatment could promote proliferationof hepatic cells in regenerating liver. Male Sprague-Dawley rats were divided into three groups: normal group (n = 3)without PH (sham operated), control groups (n = 12) with saline injection after PH, and experimental groups (n = 12) withi.p. injection of 8 mg/kg of PDRN immediately after PH. Light microscopically, liver regeneration induced by PH involvedproliferation of various hepatic cells including parenchymal cells and Kupffer cells. The disintegration (1 or 2 days afterPH) and remodeling (4 days after PH) of hepatic plate and increase of sinusoids (6 days after PH) were sequentiallyobserved. These regenerative processes occurred relatively earlier in the experimental groups. By western blotting, theexpression of PCNA increased in the early stage, reached its peak at 1 and 2 days after PH, and decreased thereafter. Byimmunohistochemistry, the intensities of PCNA staining in the experimental groups were more obvious at various daysafter PH. By electron microscopy using immunogold labeling for PCNA, the gold particles were largely observed inheterochromatin and nucleolus in hepatic cells. The results of immunochemistry and western blotting for PCNA showed thesimilar pattern in most groups. These findings suggest that PDRN treatment to hepatectomized rat could accelerate liverregeneration through rapid cell proliferation.

Hepatitis B virus inhibits the liver regeneration via epigenetic regulation of urokinase-type plasminogen activator

( Kyun Hwan Kim ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.-

The liver regeneration after liver damage caused by toxins and pathogens are critical for liver homeostasis. The retardation of liver proliferation was reported in hepatitis B virus (HBV) X protein (HBx)-transgenic mice. However, the underlying mechanism on HBx-mediated disturbance of liver regeneration was not elucidated. Here, we investigated the molecular mechanism on the inhibition of liver regeneration using liver cell lines and mouse model. Liver regeneration after partial hepatectomy (PH) was significantly inhibited in the mice model of hydrodynamic injection of HBx-expressing plasmid. Mice of hydrodynamically injected with HBx or replication-competent wtHBV plasmids showed the prominent reduction in regenerated liver weight than those of the empty vector or HBx-deficient HBV genome. Mechanism studies have revealed that the expression of urokinase-type plasminogen activator (uPA), which regulates the activation of hepatocyte growth factor (HGF), was significantly decreased in liver tissues of HBV or HBx-expressing mice. The down regulation of uPA was further confirmed using liver cell lines transiently or stably transfected with HBx and HBV genome. Furthermore, we demonstrate that HBx represses uPA expression through epigenetic regulation on uPA promoter in mice liver tissues and human liver cell lines. Expression of HBx strongly induced the hypermethylation of uPA promoter through DNMT3A2. Taken together, these data suggest that the infection of HBV can impair the liver regeneration through the epigenetic regulation of liver regeneration signal by HBx.

Hepatitis B virus inhibits the liver regeneration via epigenetic regulation of urokinase-type plasminogen activator

( Kyun Hwan Kim ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1

The liver regeneration after liver damage caused by toxins and pathogens are critical for liver homeostasis. The retardation of liver proliferation was reported in hepatitis B virus (HBV) X protein (HBx)-transgenic mice. However, the underlying mechanism on HBx-mediated disturbance of liver regeneration was not elucidated. Here, we investigated the molecular mechanism on the inhibition of liver regeneration using liver cell lines and mouse model. Liver regeneration after partial hepatectomy (PH) was significantly inhibited in the mice model of hydrodynamic injection of HBx expressing plasmid. Mice of hydrodynamically injected with HBx or replication-competent wtHBV plasmids showed the prominent reduction in regenerated liver weight than those of the empty vector or HBx-deficient HBV genome. Mechanism studies have revealed that the expression of urokinase-type plasminogen activator (uPA), which regulates the activation of hepatocyte growth factor (HGF), was significantly decreased in liver tissues of HBV or HBx-expressing mice. The down regulation of uPA was further confirmed using liver cell lines transiently or stably transfected with HBx and HBV genome. Furthermore, we demonstrate that HBx represses uPA expression through epigenetic regulation on uPA promoter in mice liver tissues and human liver cell lines. Expression of HBx strongly induced the hypermethylation of uPA promoter through DNMT3A2. Taken together, these data suggest that the infection of HBV can impair the liver regeneration through the epigenetic regulation of liver regeneration signal by HBx

Mettl14 mutation restrains liver regeneration by attenuating mitogens derived from non-parenchymal liver cells

Insook Yang,Seung Yeon Oh,Suin Jang,Il Yong Kim,You Me Sung,성제경 생화학분자생물학회 2022 BMB Reports Vol.55 No.12

Liver regeneration is a well-known systemic homeostatic phenomenon. The N6-methyladenosine (m6A) modification pathwayhas been associated with liver regeneration and hepatocellularcarcinoma. m6A methyltransferases, such as methyltransferase 3(METTL3) and methyltransferase 14 (METTL14), are involvedin the hepatocyte-specific-regenerative pathway. To illustratethe role of METTL14, secreted from non-parenchymal liver cells,in the initiation phase of liver regeneration, we performed 70%partial hepatectomy (PH) in Mettl14 heterozygous (HET) andwild-type (WT) mice. Next, we analyzed the ratio of liver weightto body weight and the expression of mitogenic stimulatorsderived from non-parenchymal liver cells. Furthermore, weevaluated the expression of cell cycle-related genes and thehepatocyte proliferation rate via MKI67-immunostaining. Duringregeneration after PH, the weight ratio was lower in Mettl14HET mice compared to WT mice. The expressions of hepatocytegrowth factor (HGF) and tumor necrosis factor (TNF)-α,mitogens derived from non-parenchymal liver cells that stimulatethe cell cycle, as well as the expressions of cyclin B1 andD1, which regulate the cell cycle, and the number of MKI67-positive cells, which indicate proliferative hepatocyte in the lateG1-M phase, were significantly reduced in Mettl14 HET mice72 h after PH. Our findings demonstrate that global Mettl14mutation may interrupt the homeostasis of liver regenerationafter an acute injury like PH by restraining certain mitogens,such as HGF and TNF-α, derived from sinusoidal endothelialcells, stellate cells, and Kupffer cells. These results providenew insights into the role of METTL14 in the clinical treatmentstrategies of liver disease.