• Journal of Ginseng Research
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• 제41권3호
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• pp.392-402
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• 2017

Background: Previously, we reported that Korean Red Ginseng inhibited liver fibrosis in mice and reduced the expressions of fibrogenic genes in hepatic stellate cells (HSCs). The present study was undertaken to identify the major ginsenoside responsible for reducing the numbers of HSCs and the underlying mechanism involved. Methods: Using LX-2 cells (a human immortalized HSC line) and primary activated HSCs, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assays were conducted to examine the cytotoxic effects of ginsenosides. $H_2O_2$ productions, glutathione contents, lactate dehydrogenase activities, mitochondrial membrane permeabilities, apoptotic cell subpopulations, caspase-3/-7 activities, transferase dUTP nick end labeling (TUNEL) staining, and immunoblot analysis were performed to elucidate the molecular mechanism responsible for ginsenoside-mediated cytotoxicity. Involvement of the AMP-activated protein kinase (AMPK)-related signaling pathway was examined using a chemical inhibitor and small interfering RNA (siRNA) transfection. Results and conclusion: Of the 11 ginsenosides tested, 20S-protopanaxadiol (PPD) showed the most potent cytotoxic activity in both LX-2 cells and primary activated HSCs. Oxidative stress-mediated apoptosis induced by 20S-PPD was blocked by N-acetyl-$\text\tiny L$-cysteine pretreatment. In addition, 20S-PPD concentration-dependently increased the phosphorylation of AMPK, and compound C prevented 20S-PPD-induced cytotoxicity and mitochondrial dysfunction. Moreover, 20S-PPD increased the phosphorylation of liver kinase B1 (LKB1), an upstream kinase of AMPK. Likewise, transfection of LX-2 cells with LKB1 siRNA reduced the cytotoxic effect of 20S-PPD. Thus, 20S-PPD appears to induce HSC apoptosis by activating LKB1-AMPK and to be a therapeutic candidate for the prevention or treatment of liver fibrosis.

• 한방재활의학과학회지
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• 제21권1호
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• pp.35-46
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• 2011

Objectives : The present study investigated inflammatory effect of Rheum undulatum L. in lipopolysaccharide-exposed rats and Raw 264.7 cells. Methods : Male rats weighting $185.39{\pm}8.21g$ fed basal diet for 1 week and 32 rats were divided into a control group and 3 experimental groups. We fed a control group of rats a basal diet and administered normal saline(100 mg/kg, 1time/1day) for 6 weeks. And we fed basal diet and administered an extract of Rheum undulatum L.(100 mg/kg, 200 mg/kg, 300 mg/kg, 1time/1day) to each experimental group of rats. We measured the plasma concentration of $IL-1{\beta}$($interleukin-1{\beta}$), IL-6 and $TNF-{\alpha}$(tumor necrosis $factor-{\alpha}$), liver cytokines, Raw 264.7 macrophages cytokines. Results : The plasma concentration of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ peaked at 5h(hour) after LPS(lipopolysaccharides) injection, and the values of the Rheum undulatum L. extract groups were lower than those of the control group. In the increment of these cytokines concentration at 2h and 5h after LPS injection, the Rheum undulatum L. groups were lower than that of control group. The plasma concentration of IL-10 peaked at 5h after LPS injection, and the values of the Rheum undulatum L. extract groups were higher than those of the control group. In the increment of this cytokine concentration at 2h and 5h after LPS injection, the Rheum undulatum L. groups were higher than that of control group. Liver cytokines measurement was done at 5h after LPS injection. The concentration of liver $IL-1{\beta}$ and IL-6 in the Rheum undulatum L. groups was lower than that of the control group. The concentrations of liver $TNF-{\alpha}$, and IL-10 showed no significant differences among all the treatment groups. In the studies of lipopolysaccharide-exposed Raw 264.7 cells, the concentration of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ in the lipopolysaccharide-exposed cells groups was higher than that of control group(normal group), and in the lipopolysaccharide-exposed cells groups, these values showed a tendency to decrease in the Rheum undulatum L. groups. The concentration of IL-10 in the lipopolysaccharide-exposed cells groups was higher than that of control group(normal group), and in the lipopolysaccharide-exposed cells groups, the values showed a tendency to increase in the Rheum undulatum L. groups. Conclusions : These results indicate that the Rheum undulatum L. extracts have an functional material for inflammatory activities.

• Nutrition Research and Practice
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• 제10권1호
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• pp.11-18
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• 2016

BACKGROUND/OBJECTIVES: Type 2 diabetes (T2D) is more frequently diagnosed and is characterized by hyperglycemia and insulin resistance. $\small{D}$-xylose, a sucrase inhibitor, may be useful as a functional sugar complement to inhibit increases in blood glucose levels. The objective of this study was to investigate the anti-diabetic effects of $\small{D}$-xylose both in vitro and stretpozotocin (STZ)-nicotinamide (NA)-induced models in vivo. MATERIALS/METHODS: Wistar rats were divided into the following groups: (i) normal control; (ii) diabetic control; (iii) diabetic rats supplemented with a diet where 5% of the total sucrose content in the diet was replaced with $\small{D}$-xylose; and (iv) diabetic rats supplemented with a diet where 10% of the total sucrose content in the diet was replaced with $\small{D}$-xylose. These groups were maintained for two weeks. The effects of $\small{D}$-xylose on blood glucose levels were examined using oral glucose tolerance test, insulin secretion assays, histology of liver and pancreas tissues, and analysis of phosphoenolpyruvate carboxylase (PEPCK) expression in liver tissues of a STZ-NA-induced experimental rat model. Levels of glucose uptake and insulin secretion by differentiated C2C12 muscle cells and INS-1 pancreatic ${\beta}$-cells were analyzed. RESULTS: In vivo, $\small{D}$-xylose supplementation significantly reduced fasting serum glucose levels (P < 0.05), it slightly reduced the area under the glucose curve, and increased insulin levels compared to the diabetic controls. $\small{D}$-xylose supplementation enhanced the regeneration of pancreas tissue and improved the arrangement of hepatocytes compared to the diabetic controls. Lower levels of PEPCK were detected in the liver tissues of $\small{D}$-xylose-supplemented rats (P < 0.05). In vitro, both 2-NBDG uptake by C2C12 cells and insulin secretion by INS-1 cells were increased with $\small{D}$-xylose supplementation in a dose-dependent manner compared to treatment with glucose alone. CONCLUSIONS: In this study, $\small{D}$-xylose exerted anti-diabetic effects in vivo by regulating blood glucose levels via regeneration of damaged pancreas and liver tissues and regulation of PEPCK, a key rate-limiting enzyme in the process of gluconeogenesis. In vitro, $\small{D}$-xylose induced the uptake of glucose by muscle cells and the secretion of insulin cells by ${\beta}$-cells. These mechanistic insights will facilitate the development of highly effective strategy for T2D.

• Applied Microscopy
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• 제34권3호
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• pp.171-178
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• 2004

메탈로사이오닌 (metallothionein: MT)은 시스테인(cysteine)이 다수 포함되어 있고, 저분자량인 단백질로서 중금속이온이나 다양한 세포 독성 인자에 의해 유도되며, 최근 발암과정 (carcinogenesis)이나 혹은 세포분화와 연관하여 활발하게 연구가 이루어지고 있다. 본 연구는 흰쥐를 수정시킨 후 발생과정 또는 출생 후 성장과정 단계에서 태아와 신생 흰쥐의 간 조직 및 세포내 MT의 분포 양상을 조사하고자 하였다. 흰쥐 간 조직내 MT의 분포는 면역조직화학적 방법을 적용하여 광학현미경으로 관찰하였고, 세포내 미세구조적 위치 (localization)는 금 입자 표지를 이용한 면역세포화학적 방법을 이용하여 투과전자현미경으로 관찰하였다. MT는 발생 13일이 경과한 흰쥐 태아의 간에서 처음으로 관찰되기 시작하여, 출생 당일의 신생 흰쥐에서 가장 높게 관찰되었다. 면역 조직화학 및 세포화학적 방법으로 관찰한 결과, MT 염색성은 발생 후기에서 출생후 10일째까지 비교적 높게 나타났다. MT에 대한 양성반응은 난형 세포 (oval cell)들과 간 실질세포 (liver parenchymal cell)들에서 뚜렷하게 존재하였다. 발생과정과 신생 흰쥐의 간 조직에서는 핵과 세포질 양쪽에서 공통적으로 양성 반응을 보였으나, 생후 30일 이후부터 성체에 이르는 시기까지는 세포질에 한정되어 있는 양상을 보였다. 금 입자 표지법에 의한 MT의 세포질내 분포를 보면, 발생 중인 흰쥐 간 세포에서는 핵과 세포질내에 고르게 산재되어 있었다. 즉, 핵내 염색질, 인, 조면소포체 및 cytosol에서도 MT에 대한 면역 금 입자들이 관찰되었으나, 미토콘드리아와 지질 소포 등에서는 거의 관찰되지 않았다. 상기의 결과들은 발생중 혹은 성장중인 흰쥐의 간에서 MT가 점차 증가하는 것으로 요약될 수 있는 것으로서, 이는 MT가 세포들의 증식과 분화에 금속 이온의 저장과 방출 혹은 전사인자 (transcription factor)로서 작용할 수 있음을 암시하는 것으로 판단된다.

• Biomolecules & Therapeutics
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• 제16권4호
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• pp.306-311
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• 2008

Although hepatic microcirculatory dysfunction occurs during endotoxemia, the mechanism responsible for this remains unclear. Since Kupffer cells provide signals that regulate hepatic response in inflammation, this study was designed to investigate the role of Kupffer cells in the imbalance in the expression of vasoactive mediators. Endotoxemia was induced by intraperitoneal E. coli endotoxin (LPS, 1 mg/kg body weight). Kupffer cells were inactivated with gadolinium chloride ($GdCl_3$, 7.5 mg/kg body weight, intravenously) 2 days prior to LPS exposure. Liver samples were taken 6 h following LPS exposure for RT-PCR analysis of mRNA for genes of interest: endothelin (ET-1), its receptors $ET_A$ and $ET_B$, inducible nitric oxide synthase (iNOS), heme oxygenase (HO-1), and tumor necrosis factor-$\alpha$ (TNF-$\alpha$). mRNA levels for iNOS and TNF-$\alpha$ were significantly increased 31.8-fold and 26.7-fold in LPS-treated animals, respectively. This increase was markedly attenuated by $GdCl_3$, HO-1 expression significantly increased in LPS-treated animals, with no significant difference between saline and $GdCl_3$ groups. ET-1 was increased by LPS. mRNA levels for $ET_A$ receptor showed no change, whereas $ET_B$ transcripts increased in LPS-treated animals. The increase in $ET_B$ transcripts was potentiated by $GdCl_3$. We conclude that activation of Kupffer cells plays an important role in the imbalanced hepatic vasoregulatory gene expression induced by endotoxin.

• 한국동물위생학회지
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• 제29권3호
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• pp.347-364
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• 2006

This study is concerned with assessment of diethylnitrosamine (DEN 0.01 %) induced liver cell carcinogenesis by measurement of changes preceding the development of neoplasms. Therefore, it was undertaken to investigate changes of liver-specific enzyme activities in Sprague-Dawley (SD) rats by ad libitum feeding of DEN. And also. the changes of hepatic morphology in SD rats were detected by haematoxylineosin stain and immunohistochemistry (PCNA). 5- Fluorouracil (5- FU) is one of the most widely used anticancer agents for digestive cancers including hepatocellular carcinoma, and is known to affect the cell cycle and induce apoptosis of cancer cells. In the present study, SD rats were given drinking water containing 0.01% diethylnitrosamine (DEN) for 8 weeks. Minor behavioral change, brittleness of hair and decreased amount of water and diet intake were observed in rats 4 weeks after DEN administration. The body and liver weights were significantly (p < 0.05) decreased in rats 11 weeks after DEN administration. The liver weight ratio to body weight was rather stable and not significantly decreased in the all treatment groups. The liver specific enzyme activities (AST, ALT, ${\gamma}$-GTP) were significantly increased in all treatment groups compared to control group (p < 0.05). Variable size of liver tumor and hepatomegaly were observed in rats treated with DEN after 10 weeks. Numerous vacuoles were seen on the midzonal and or peripheral areas of hepatic lobules. The large and polymorphological hepatocytes with eosinophilic cytoplasm or densely basophilic mitotic nucleoli were seen. Several proliferative small round cells were seen on vacuolated and necrotic areas in peripheral hepatic lobules or portal areas. PCNA-positive cells were seen on the vacuolated portal areas and peripheral areas of hepatic lobules in the areas of small round cells. We examined functional and morphological changes of livers by 5 - FU treatments on DEN -treated rat. The DEN -treated rats compared to 5 - FU -treated rats after DEN treatment for 8 weeks. The serum total protein and triglyceride were significantly (p < 0.05) decreased, and the liver enzyme activities of AST and ALT were significantly(p < 0.05) increased. After 8 weeks, in the non-5-FU -treated group, the size of liver tumor were varied and hepatomegaly were observed, hepatocellular vacuolization, necrosis and steatosis were observed on the midzonal and peripheral areas of hepatic lobules. The large and polymorphological hepatocytes were seen, the interlobular connective tissues were proliferated. PCNA positive cells were seen in the portal areas and peripheral areas of hepatic lobules in the non-5-FU-treated group. In hepatocytes, condensation of nuclear chromatin and vacuolization were observed, shape of the nuclei were irregular, the degraded nuclei and organelles were observed. The livers of rats in the 5 - FU treatment group were seen grossly brilliant, red-brown color, and the vacuolated and degenerated regions, hyperplastic nodules were not nearly observed. In the electron microscope, the cytoplasm of the hepatocytes contained a large number of mitochondria, rough endoplasmic reticulum, developed organelles surrounding nuclei. The above findings suggest that 5 - FU will be effective as anti -liver tumor drug.

• 한국동물학회지
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• 제16권2호
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• pp.79-96
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• 1973

붕어(Carassius carassius)에 미치는 동의 영향을 밝히기 위하여 다음과 같은 연구를 하였다. 1) 셀루로오스 아세테이트 전기영동법에 의한 젖산수소이탈효소 아이소자임 상, 2) 분광비색법에 의한 LDH활성과 LDH효소계에 대한 동의 영향, 3) 한천 박층 전기영동법에 의한 에스테라아제 아이소자임 상, 4) 전분 겔 전기영동법에 의한 혈색소 상, 및 5) 조직 학적 연구. 1. LDH 아이소자임 영동대는 정상 붕어의 아가미에 2개 (LDH-3 및 LDH-5), 간에 3개(LDH-2, LDH-4, 및 LDH-5), 그리고 근육에서 2개 (LDH-3 및 LDH-4) 나타났다. LDH-1은 이상의 세가지 조직에서 나타나지 않았다. 동을 처리한 붕어의 간에서는 LDH-3이 나타났고 근육에서는 LDH-5가 나타났다. 그러나 아가미에서는 새로운 영동대가 나타나지 않았다. 2. LDH활성은 정상 붕어의 아가미, 간, 및 근육 중 아가미에서 가장 낯았고 근육에서 가장 높았으며, 아가미를 제외한 간과 근육에서는 1일부터 10일간의 동처리에 따라 점차적으로 감소되었다. 3. 동을 처리한 붕어의 간과 근육의 LDH활성 감소는 주로 체내 M-LDH에 대한 동의 억제에 기인된다. 4. 정상 붕어의 아가미, 간, 근육, 혈액, 뇌, 및 신장의 에스테라아제 아이소자임 영동대 수는 각각 3,6,2,2,2 및 2개 였고 이들은 동을 처리한 경우에서도 같았다. 동처리군의 아가미, 간, 혈액 그리고 신장의 에스테라아제 영동대의 상대이동도는 대조군의 그것들과 상이하였다. 5. 정상 붕어의 혈색소 영동대는 양극에 1개 있었다. 동을 처리한 붕어의 혈색소 영동대의 이동도는 정상 붕어의 그 것보다 약간 빠른 것으로 보였다. 6. 동(20ppm)에 5일간 처리한 붕어의 조직학적 연구는 다음과 같았다. 1) 아가미에서는 처음에 표피층이 분리되었다가 분해된 후 완전히 파괴되었다. 2) 간에서는 많은 지방립이 나타났다. 3) 정상 붕어와 동에 처리된 붕어의 근육 사이에서는 조직학적 차이가 없었다. 4) 신장족직은 기부요세관에 손상을 받았다. 요세관세포의 높이가 작아진 후 요세관의 내강이 확대되었다. 많은 기부요세관이 분홍빛의 과립성 형태와 여러 단계의 변성을 나타내었다.

• Nutrition Research and Practice
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• 제12권2호
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• pp.110-117
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• 2018

BACKGROUND/OBJECTIVES: Non-alcoholic fatty liver disease (NAFLD) is a leading cause of chronic liver disease and is closely associated with metabolic syndrome. In the present study, we observed the effect of ethanol extract of Allium fistulosum (EAF) on NAFLD and have suggested the possibility of using EAF as a natural product for application in the development of a treatment for NAFLD. MATERIALS/METHODS: The preventive effect on hepatic lipid accumulation was estimated by using an oleic acid (OA)-induced NAFLD model in vitro and a Western diet (high-fat high-sucrose; WD)-induced obese mouse model. Animals were divided into three groups (n = 7): normal diet group (ND), WD group, and WD plus 1% EAF group. RESULTS: EAF reduced OA-stimulated lipid accumulation in HepG2 cells in the absence of cellular cytotoxicity and significantly blocked transcriptional activation of sterol regulatory element-binding protein 1 and fatty acid synthase genes. Subsequently, we investigated these effects in vivo in mice fed either ND or WD in the presence or absence of EAF supplementation. In comparison to the ND controls, the WD-fed mice exhibited increases in body weight, liver weight, epididymal fat weight, and accumulation of fat in hepatocytes, and these effects were significantly attenuated by EAF supplementation. CONCLUSIONS: Allium fistulosum attenuates the development of NAFLD, and EAF elicits anti-lipogenic activity in liver. Therefore, EAF represents a promising candidate for use in the development of novel therapeutic drugs or drug combinations for the prevention and treatment of NAFLD.

• 대한융합한의학회지
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• 제5권1호
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• pp.45-54
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• 2023

Objectives : Alcohol-induced liver disease advances as to reactive oxygen species (ROS) and cellular lipid peroxidation increase. We examined the hepatoprotective effects of Zingiber officinale Roscoe rhizome extract (ZR), Pueraria lobata Ohwi flower extracts (PF), and a newly developed herbal juice (HJ), which was a combination of ZR and PF extracts, against ethanol-induced hepatotoxicity. Methods: The study utilized the human hepatoma cell line HepG2 cells to validate the hepatoprotective effect of HJ (50~200 ㎍/mL) against ethanol (EtOH, 700 mM)-induced liver damage. Results: HJ effectively reduced the protein expression of sterol regulatory element-binding transcription factor 1, adiponectin, and AMP-activated protein kinase in EtOH-induced HepG2 cells. The levels of ROS, total cholesterol, and triglycerides, which are the result of various synthesis and lipogenesis processes induced by EtOH in the liver, were reduced by HJ. Furthermore, the activities of alcohol dehydrogenase and aldehyde dehydrogenase, enzymes linked to alcohol degradation, were more effectively downregulated by HJ treatment compared to treatment with ZR and PF alone, all without causing cytotoxic effects. Conclusions: HJ protects the liver by inhibiting EtOH-induced lipogenesis, lowering ROS generation, and improving alcohol degradation, which is more effective than ZR and PF alone. Further, in vivo experiments can offer additional evidence regarding the effectiveness, safety, and underlying mechanism of action of HJ.

• Parasites, Hosts and Diseases
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• 제62권1호
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• pp.30-41
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• 2024

The dense granule protein of Toxoplasma gondii, inhibitor of signal transducer and activator of transcription 1 (IST) is an inhibitor of signal transducer and activator of transcription 1 (STAT1) transcriptional activity that binds to STAT1 and regulates the expression of inflammatory molecules in host cells. A sterile inflammatory liver injury in pathological acute liver failures occurs when excessive innate immune function, such as the massive release of IFN-γ and TNF-α, is activated without infection. In relation to inflammatory liver injury, we hypothesized that Toxoplasma gondii inhibitor of STAT1 transcription (TgIST) can inhibit the inflammatory response induced by activating the STAT1/IRF-1 mechanism in liver inflammation. This study used IFN-γ and TNF-α as inflammatory inducers at the cellular level of murine hepatocytes (Hepa-1c1c7) to determine whether TgIST inhibits the STAT1/IRF-1 axis. In stable cells transfected with TgIST, STAT1 expression decreased with a decrease in interferon regulatory factor (IRF)-1 levels. Furthermore, STAT1 inhibition of TgIST resulted in lower levels of NF-κB and COX2, as well as significantly lower levels of class II transactivator (CIITA), iNOS, and chemokines (CLXCL9/10/11). TgIST also significantly reduced the expression of hepatocyte proapoptotic markers (Caspase3/8/9, P53, and BAX), which are linked to sterile inflammatory liver injury. TgIST also reduced the expression of adhesion (ICAM-1 and VCAM-1) and infiltration markers of programmed death-ligand 1 (PD-L1) induced by hepatocyte and tissue damage. TgIST restored the cell apoptosis induced by IFN-γ/TNF-α stimulation. These results suggest that TgIST can inhibit STAT1-mediated inflammatory and apoptotic responses in hepatocytes stimulated with proinflammatory cytokines.