Chunxue Li ;Yating Zhan ;Rongrong Zhang;Qiqi Tao ;Zhichao Lang ;Jianjian Zheng
Journal of Ginseng Research
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v.47
no.4
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pp.515-523
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2023
Background: 20(S)-protopanaxadiol (PPD), one of the main components of ginseng, has anti-inflammatory, anti-estrogenic, and anti-tumor activities. It is known that activated hepatic stellate cells (HSCs) are the primary producers of extracellular matrix (ECM) in the liver, and the Wnt/β-catenin pathway participates in the activation of HSCs. We aimed to explore whether PPD inhibits liver fibrosis is associated with the Wnt/β-catenin pathway inactivation. Methods: The anti-fibrotic roles of PPD were examined both in vitro and in vivo. We also examined the levels of Wnt inhibitory factor 1 (WIF1), DNA methyltransferase 1 (DNMT1) and WIF1 methylation. Results: PPD obviously ameliorated liver fibrosis in carbon tetrachloride (CCl4)-treated mice and reduced collagen deposition. PPD also suppressed the activation and proliferation of primary HSCs. Notably, PPD inhibited the Wnt/β-catenin pathway, reduced TCF activity, and increased P-β-catenin and GSK-3β levels. Interestingly, WIF1 was found to mediate the inactivation of the Wnt/β-catenin pathway in PPD-treated HSCs. WIF1 silencing suppressed the inhibitory effects of PPD on HSC activation and also restored α-SMA and type I collagen levels. The downregulation of WIF1 expression was associated with the methylation of its promoter. PPD induced WIF1 demethylation and restored WIF1 expression. Further experiments confirmed that DNMT1 overexpression blocked the effects of PPD on WIF1 expression and demethylation and enhanced HSC activation. Conclusion: PPD up-regulates WIF1 levels and impairs Wnt/β-catenin pathway activation via the downregulation of DNMT1-mediated WIF1 methylation, leading to HSC inactivation. Therefore, PPD may be a promising therapeutic drug for patients with liver fibrosis.
Journal of the Korean Society of Food Science and Nutrition
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v.46
no.11
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pp.1278-1285
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2017
Accumulation of excess low density lipoprotein (LDL) cholesterol in the blood can initiate and accelerate atherosclerosis. Statins mediate the transactivation of proprotein convertase subtilisin/kexin type 9 (PCSK9), which in turn limits their cholesterol-lowering effects via LDL receptor (LDLR) degradation. The objective of this study was to investigate whether or not Allium tuberosum (AT) regulates LDLR and PCSK9. Mice were fed a low fat control diet (LD) or Western diet (WD) supplemented with AT (1%, w/w). AT significantly attenuated total and LDL cholesterol levels in mice fed WD (P<0.05). AT also significantly inhibited hepatic PCSK9 gene expression (P<0.05) while AT maintained hepatic LDLR gene expression. To further investigate AT-mediated PCSK9 regulation, HepG2 cells were treated with 10% delipidated serum (DLPS) in the presence or absence of AT. Non-toxic level of AT dose-dependently increased the LDLR protein level, and AT at $400{\mu}g/mL$ markedly inhibited PCSK9 protein expression. Similarly, AT significantly increased LDLR gene expression, whereas it significantly down-regulated PCSK9 gene expression. AT-mediated reduction of PCSK9 gene expression is likely due to decreased hepatic nuclear factor $1{\alpha}$ ($HNF1{\alpha}$) expression, but not SREBP2 in HepG2 cells under lipid-depleted conditions. AT-mediated PCSK9 inhibition contributed to LDLR protein stabilization via protection against LDLR lysosomal degradation in HepG2 cells under lipid-depleted conditions. Further investigation is warranted to determine the active components of AT and whether or not these components are effective in reducing hypercholesterolemia.
The objective of this research was to study the effect of astaxanthin (AST) on growth performance and antioxidant capacity in golden pompano (Trachinotus ovatus) both in vivo and in vitro. In the in vivo study, two diets were formulated with or without astaxanthin supplementation (D1 and D2; 0 and 200 mg/kg) to feed fish for 6 weeks. In the in vitro study, cells from hepatopancreas of golden pompano were isolated and four treatments with or without astaxanthin and $H_2O_2$ supplementation were applied (control group: without both astaxanthin and $H_2O_2$ treated; $H_2O_2$ group: just with $H_2O_2$ treated; $H_2O_2$ + AST group: with both astaxanthin and $H_2O_2$treated; AST group: just with AST treated). Results of the in vivo study showed that weight gain (WG) and special growth rate (SGR) significantly increased with astaxanthin supplemented (P < 0.05). Feed conversion ratio (FCR) of fish fed D2 diet was significantly lower than that of fish fed D1 diet (P < 0.05). Hepatic total antioxidant capacity (T-AOC) and the reduced glutathione (GSH) of golden pompano fed D2 diet were significant higher than those of fish fed D1 diet (P < 0.05). Superoxide dismutase (SOD) was significantly declined as astaxanthin was supplemented (P < 0.05). Results of the in vitro study showed that the cell viability of $H_2O_2$ group was 52.37% compared to the control group, and it was significantly elevated to 84.18% by astaxanthin supplementation ($H_2O_2$ + AST group) (P < 0.05). The total antioxidant capacity (T-AOC) and the reduced glutathione (GSH) of cell were significant decreased by oxidative stress from $H_2O_2$ (P < 0.05), but it could be raised by astaxanthin supplementation ($H_2O_2$ vs $H_2O_2$ + AST), and the malondialdehyde (MDA) was significant higher in $H_2O_2$ group (P < 0.05) and astaxanthin supplementation could alleviate the cells from lipid peroxidation injury. In conclusion, dietary astaxanthin supplementation can improve the growth performance of golden pompano. Moreover, astaxanthin can improve the golden pompano hepatic antioxidant capacity both in vivo and in vitro study by eliminating the reactive oxygen species.
Hepatitis B viral infection which affect about 10% of Korean population manifests asymptomatic carrier, chronic hepatitis and liver cirrhosis and even associates with hepatocellular carcinoma. Clinical manifestations induced by hepatitis B virus vary depending on the degree of immune response by cytotoxic T cells against viral epitope-presenting liver cells. Since hepatitis B virus presents high rate of mutaton that might change the presented epitope and eventually alter immune response, viral mutations, especially in promoters and enhancers, have an important implication in hepatic inflammation and viral replication. To identify mutations related to the hepatic inflammation, we investigated sequence variations of hepatitis B viral promotor regions in the presence or absence of symptoms in hepatitis B carriers. For this, sera from persistently hepatitis B virus-infected mother-child pairs were collected. After PCR amplifiation of all hepatitis B viral promoters (C promoter, S1 promoter, S2/S promoter, X promoter) using serum DNA from each pair, viral promotors were sequenced by automatic sequencer and then sequence data were analyzed by ClustalW. In most cases, the dominant type of maternal virus was transmitted to the child. However, in some children, some new host specific viral variants could be observed in Cp, S1p and S2/Sp. The mutations in C promoter did not seem to be vertically transmitted but arose in new host independently after the wild type had been transmitted. Enhancer I containing X promoter revealed high host specific variations as has been reported before. Two S promoters, S1p and S2/Sp, have shown some point mutations in children, but no deletion mutations were detected as in chronic hepatitis patients in whom deletion mutations are frequently found. In conclusion, the children with the vertically transmitted hepatitis B virus mostly retain the dominant type virus that had been transmitted. However, host specific variants tended to accumulate over time, possibly as clinical symptoms develop.
Objectives : In this study, effects of haepyoijintang (HIJ) on the increase in airway epithelial mucosubstances of rats and ATP-, PMA-, EGF- or TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells were investigated. Methods : Hypersecretion of airway mucus was induced by exposure of rats to $SO_2$ during 3 weeks. Effect of orally-administered HIJ during 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats was evaluated using histopathological analysis after staining the epithelial tissue with PAS-alcian blue. Possible cytotoxicity of HIJ was evaluated by examining the potential damage of kidney and liver functions by measuring serum GOT/GPT activities and serum BUN and creatinine concentrations of rats and the body weight gain during experiment, after administering HIJ orally. At the same time, the effect of HIJ on ATP-, PMA-, EGF- or TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of HIJ and treated with ATP ($200{\mu}M$), PMA (10 ng/ml), EGF (25 ng/ml) or TNF-${\alpha}$ (0.2 nM) for 24 hrs, to evaluate the effect of HIJ both on ATP-, PMA-, EGF- or TNF-${\alpha}$-induced MUC5AC mucin production using enzyme-linked immunosorbent assay (ELISA) and on gene expression by the same inducers using reverse transcription-polymerase chain reaction (RT-PCR). Results : (1) HIJ decreased the amount of intraepithelial mucosubstances of trachea of rats. (2) HIJ did not show renal and hepatic toxicities and did not affect body weight gain of rats during experiment. (3) HIJ significantly inhibited ATP-, PMA-, EGF-, and TNF-${\alpha}$-induced MUC5AC mucin productions from NCI-H292 cells. (4) HIJ significantly inhibited ATP-, PMA-, EGF-, and TNF-${\alpha}$-induced MUC5AC mucin gene expression from NCI-H292 cells. Conclusions : The result from the present study suggests that HIJ might control the production and gene expression of airway mucin observed in various respiratory diseases accompanied by mucus hypersecretion and do not show in vivo toxicity to liver and kidney functions after oral administration. Effect of HIJ with their diverse components should be further investigated using animal experimental models that can reflect the pathophysiology of airway diseases through future studies.
The present study was designed to investigate i) the action of various nucleotides on membrane permeability of rat red blood cell and hepatocyte for $Na^{+}$ and $Rb^{+}$ ii) the characteristics of purinoceptors on these cell membranes. Blood from Sprague-Dawley rats was obtained by carotid arterial cannulation. Red blood cells were then washed 3 times with saline at $4{\circ}C$. Hepatic parenchymal cells were isolated from rat livers by using a modification of the Berry and Friend (1969) method. For the $Na^{+}$ influx studies, isolated RBC and hepatocyte were incubated in incubation medium containing $^{22}Na^{+}0.2\;{\mu}Ci/ml$ at $37^{\circ}C$. After various time intervals samples were removed from the incubation flask and washed out 3 times with ice-cold washing solutions. Cells were destroyed by adding Triton X-100 and TCA solution. After centrifugation, the supernatants were assayed for $^{22}Na^{+}$ by gamma counter. $^{86}Rb^{+}$ was used to simulate $K^{+}$ in these $K^{+}efflux$ studies. Isolated hepatocytes were incubated for 60 min in the loading solution containing $^{86}Rb^{+}\;10\;{\mu}Ci/ml$ at $37^{\circ}C$. After loading, the cells washed out 3 times by centrifugation with washing solution. The cells were incubated in buffer solution at $37^{\circ}C$. At intervals thereafter, samples were removed and centrifuged. The supernatants were analyzed for $^{86}Rb^{+}$ by liquid scintillation counter. The main results of the experiments were: 1) ATP and ATPP increased in both $^{22}Na^{+}$ influx and $^{86}Rb^{+}$ efflux in the red blood cell. Although ADP showed a tendency to increase in RBC membrane permeability for $^{22}Na^{+}$ and $^{86}Rb^{+}$, the changes were not significantly different from the control. 2) The Significant changes in $^{22}Na^{+}$ and $^{86}Rb^{+}$ flux by ATP were also demonstrated in hepatocyte. ATPP and ADP showed a tendency to increase in hepatocyte membrane permeability for both ions. 3) Other nucleoside triphosphates-ITP, GTP and CTP-did not change in membrane permeability for $^{22}Na^{+}$ and $^{86}Rb^{+}$ in RBC and hepatocyte. In conclusion, not only ATP but also ATPP activate purinoceptors and change in membrane permeability for $Na^{+}$ and $K^{+}$. In order to activate purinoceptors on the cell membrane, the nucleotides have to possess intact adenine moiety and three phosphates or more in its molecule.
Journal of the Korean Society of Food Science and Nutrition
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v.36
no.1
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pp.8-13
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2007
This study was conducted to investigate the effect of mycelia of Lentinus edoes mushroom-cultured Astragalus membranaceus Bunge (LAM) on proliferation of cancer cell lines (Hep3B, MCF-7 and HeLa), sarcoma 180 (S-180), and anti-allergy. In an anti-cancer test using Hep3B (hepatic cancer cell), MCF-7 (breast cancer cell) and HeLa (uterine cancer cell), LAM extract showed higher antiproliferating effect than that of Astragalus membranaceus Bunge (AM) extract. In an anti-cancer testing using Hep3B cells and MCF-7 cells, LAM extract showed growth-inhibitory effect of 65.23% at 3 mg/mL and 69.23% at 5 mg/mL, respectively. In an anti-cancer testing using HeLa cells, LAM extract showed growth-inhibitory effect of 42.01% at 5 mg/mL. In addition, LAM showed the tumor suppressive effect in mice injected with S-180 cells. The growth-inhibitory rates against tumor cells were 47% for LAM and 37% for AM. LAM inhibited histamine release from rat peritoneal mast cells activated by compound 48/80. These results suggest that Lentinus edodes mushroom-cultured herb has an antiproliferating effect against cancer cell lines (Hep3B, MCF-7 and HeLa), S-180 tumor and will be beneficial in the treatment of allergic reaction.
Liver regeneration is a result of highly coordinated proliferation of hepatocytes and nonparenchymal liver cells. Partial hepatectomy (PH) is the most often used stimulus to study liver regeneration because, compared with other methods that use hepatic toxins, it is not associated with the tissue injury and inflammation, and the initiation of the regenerative stimulus is precisely defined. Granulocyte macrophage-colony stimulating factor (GM-CSF), which is a cytokine able to regulate the proliferation and differentiation of epithelial cells, was first identified as the most potent mitogen for bone marrow. Particularly, rrhGM-CSF, which is highly glycosylated and sustained longer than any other types of GM-CSF in the blood circulation, was specifically produced from rice cell culture. In this experiment, effects of rrhGM-CSF administration were evaluated in the regenerating liver after 78% PH of rats. Morphological changes induced by PH were characterized by destroyed hepatocyte plate around the central vein and enlarged nuclear cytoplasmic ratio and increased hepatocytes with two nuclei. And then, proliferation of liver cells (parenchymal and nonparenchymal) and rearrangement of plates and lobules seemed to be carried out during liver regeneration. These alterations in the experimental group preceded those of the control. Since proliferating cell nuclear antigen (PCNA) is known to be a nuclear protein maximally elevated in the S phase of proliferating cells, the protein was used as a marker of liver regeneration after PH in rats. PCNA levels by western blot analysis and immunohistology were compared between the two groups. PCNA protein expression of two groups at 12 hr and 24 hr after injury showed similar pattern. The protein expression showed the peak at 3 days in both groups, however, the protein level of the experimental group was higher than that of the control. On immunohistochemical observations, the reaction product of PCNA was localized at the nuclei of proliferating cells and the positive reaction in experimental group at 3 days was clearly stronger than that in control group. The results by Western blotting and immunohistology for PCNA showed similar pattern in terms of the protein levels. In conclusion, rrhGM-CSF administration during liver regeneration after 78% PH accelerated breakdown and restoration of the hepatic plate and expression of PCNA. These results suggest that rrhGM-CSF might play an important role during liver regeneration in rats.
Journal of the Korean Society of Food Science and Nutrition
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v.36
no.4
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pp.424-430
/
2007
This study was conducted to investigate the effect of mycelia of Lentinus edoes mushroom-cultured Lonicera japonica Thunberg (LLJ) on proliferation of the cancer cell lines (Hep3B, MCF-7 and HeLa), sarcoma 180 (S-180) and antiallergy. In an anti-cancer test using Hep3B (hepatic cancer cell), MCF-7 (breast cancer cell) and HeLa (uterine cancer cell), LLJ extract showed higher antiproliferating effect than that of LJ (Lonicera japonica Thunberg) extract. In an anti-cancer testing using Hep3B cells, LLJ extract showed growth-inhibitory effect of $85.60{\pm}4.66%$ at 3mg/rnL. In an anti-cancer testing using MCF-7 cells, LLJ and LJ extracts showed high antiproliferating effect. LLJ showed the tumor suppressive effect in mice injected with S-180 cells. The growth-inhibitory rates against tumor cells were 61% for LLJ, 37% for LJ. LLJ inhibited histamine release from rat peritoneal mast cells activated by compound 48/80. These results suggest that Lentinus edodes mushroom-cultured herb has an antiproliferating effect against cancer cell lines (Hep3B, MCF-7 and HeLa) and S-180 tumor, and will be beneficial in the treatment of allergic reaction.
The hypothesis that calcium provoke $O_2^-$ formation by Kupffer cells and may contribute to carbon tetrachloride $(CCl_4)$ induced liver injury was studied in SD rats. In $CCl_4-treated$ animals, hepatic malonaldehyde (nmole/gm liver) and plasma ALT (IU/ml) levels elevated significantly from $119.63{\pm}13.00$ to $268.97{\pm}14.82$ and from $17.3{\pm}0.18$ to $806.08{\pm}37.63$, respectively, compared to those in controls. Activation of Kupffer cells with high dose of retinol (250,000 IU/kg/day, po, for 7 day) significantly enhanced ALT levels, while inactivation of Kupffer cells with gadolinium chloride (7.5 mg/kg/day, ip, for 2 day) attenuated the increase of serum ALT level following $CCl_4$ treatment. Diltiazem (10 mg/kg/day, ip for 2 day) given in combination with retinol led to a marked decrease in ALT levels compare to the level in rats treated only with retinol against $CCl_4$ treatment. In order to determine any alterations in cytochrome P450 activities, the P450 content and the CYP2E1 activity were measured and all $CCl_4-treated$ rats showed significantly lower levels compared to those in controls and vehicle-treated animals. There were significant increases in glutathione peroxidase in all $CCl_4-treated$ rats except diltiazem treated groups. No difference was found among untreated and vehicle-treated rats. It is concluded that Kupffer cells contribute to $CCl_4-induced$ liver injury and that calcium antagonist attenuated the increased $CCl_4-induced$ liver injury due to activation of Kupffer cells.
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