• Toxicological Research
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• 제11권2호
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• pp.181-185
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• 1995

To know the mechanism of biphenyl dimethyl dicarboxylate (DDB) in the protection of chemically induced hepatotoxicity, the activity of glutamic pyruvic tran.saminase (GPT) and the level of lipid peroxidation metabolite (malondialdehyde, MDA) and ATP content in hepatocytes were determined in serum and primarily cultured hepatocytes. For in vibo study, rats were pretreated with DDB (300 mg/ kg, p.o.)for 7 days. DDB pretreatment efficiently reduced the elevation of serum GPT activity induced by carbon tetrachloride (1.6 ml/kg, s.c.) and acetaminophen administration (1500 mg/kg, i.p.). In ex vivo study, hepatocytes were isolated from the rats pretreated with DDB (300 mg/kg, p.o.)for 7 days and cultured for 12 hrs before inducing cytotoxicity with chemicals. The MDA formation and the GPT release induced by adriamycin $(1\times10^{-4} mg/ml)$ and cisplatin $(2\times10^{-4} mg/ml)$ were markedly decreased in the hepatocytes from the rats pretreated with DDB as compared to vehicle only. However, DDB pretreatment did not prevent the decrease of ATP contents of hepatocytes induced by cisplatin and adriamycin. In in vitro experiment, DDB was pretreated in primary cultured hepatocytes for 3 days. DDB enhanced the decreases of ATP contents induced by cisplatin and adriamycln. These results suggest that DDB may protect the hepatocytes from injury induced by hepatotoxlcants through inhibiting the lipid peroxidation.

• Fisheries and Aquatic Sciences
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• 제1권1호
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• pp.19-23
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• 1998

Effects of pituitary and thyroid hormones on estradiol-induced vitellogenin (VTG) induction were electrophoretically examined in primary hepatocyte cultures of rainbow trout. Hepatocytes were precultured for 2 days and then estradiol-17 $\beta$ $(E_2,\;2 \times 10^{-6}M)$>, triiodothyronine $(T_3,\;10^{-8}-10^{-6}M)$, bovine growth hormone (bGH, 10-100 ng/ml), ovine prolactin (oPRL, 100-500 ng/ml), and pituitary extract (PE) of rainbow trout (0.75PE/dish) were added to the incubation medium. The hepatocytes were cultured for 7 more days. The addition of oPRL to the incubation medium was not effective in increasing VTG production at any concentrations. The addition of PE to the incubation medium with $E_2$ was not effective in increasing VTG production. The addition of bGH to the incubation medium with $E_2$ was not effective in increasing the rate of VTG production at concentrations of 10-50 ng/ml. However, a higher concentration of bGH, 100 ng/ml, increased VTG production. The various concentrations of $T_3$ were ineffective in stimulating VTG production. These results suggest that GH could be one of stimulus factors for VTG production in rainbow trout.

• Journal of Animal Science and Technology
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• 제65권1호
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• pp.16-31
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• 2023

Cultured meat is a potential sustainable food generated by the in vitro myogenesis of muscle satellite (stem) cells (MSCs). The self-renewal and differentiation properties of MSCs are of primary interest for cultured meat production. MSC proliferation and differentiation are influenced by a variety of growth factors such as insulin-like growth factors (IGF-1 and IGF-2), transforming growth factor beta (TGF-β), fibroblast growth factors (FGF-2 and FGF-21), platelet-derived growth factor (PDGF) and hepatocyte growth factor (HGF) and by hormones like insulin, testosterone, glucocorticoids, and thyroid hormones. In this review, we investigated the roles of growth factors and hormones during cultured meat production because these factors provide signals for MSC growth and structural stability. The aim of this article is to provide the important idea about different growth factors such as FGF (enhance the cell proliferation and differentiation), IGF-1 (increase the number of myoblasts), PDGF (myoblast proliferation), TGF-β1 (muscle repair) and hormones such as insulin (cell survival and growth), testosterone (muscle fiber size), dexamethasone (myoblast proliferation and differentiation), and thyroid hormones (amount and diameter of muscle fibers and determine the usual pattern of fiber distributions) as media components during myogenesis for cultured meat production.

• 한국어류학회지
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• 제12권3호
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• pp.180-185
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• 2000

플라스틱의 원재료로 이용되는 bisphenol-A (BPA)가 vitellogenin (VTG)의 합성에 미치는 영향을 넙치의 간세포를 초대 배양하여 조사하였다. 간세포는 2일간 사전 배양을 하였으며, estradiol-$17\beta(E_2)$ 및 BPA를 배양액에 동시 첨가하였다. 간세포는 6일간 배양한 후 배양액을 회수하여 SDS-PAGE로 분석하였다. BPA는 농도 의존적으로 총 단백질에 대한 VTG 비율은 증가하였으며, $10^{-6}M$과 $10^{-5}M$의 농도에서 유의한 차이를 나타내었다(P<0.05). 특히, BPA $10^{-5}M$의 농도에서의 총 단백질에 대한 VTG 비율은 26.36%로 $E_2$만을 첨가한 대조군과 유의한 차이를 나타내지 않았다. $E_2$와 BPA를 단독 또는 동시에 첨가하여 유도된 VTG합성은 배양액에 tamoxifen$(10^{-6}M)$을 첨가함으로서 $E_2$를 첨가한 대조군의 약 80%로 유의하게 억제되었다. 그리고, in vivo에서 $E_2$에 의해 유도된 VTG합성은 in vitro에서 간세포를 6일간 배양함에 따라 $E_2$를 첨가한 대조군의 약 22%까지 억제되었다. 이러한 감소효과는 BPA의 농도 의존적으로 지연되는 결과를 나타내었다. 따라서 넙치에서의 BPA는 $E_2$ receptor와의 반응을 통한 에스트로겐 작용에 의해 VTG합성을 유도하고, 난황형성기에서의 VTG 합성을 장시간 지속시킬 가능성이 있는 것으로 추측된다.

• Toxicological Research
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• 제23권4호
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• pp.301-309
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• 2007

Paecilomyces tenuipes (PT), one of the Ascomycetes family, has been used for medicinal purposes due to its broad pharmacological activities. The present study was undertaken to investigate the hepatoprotective effects of PT water extracts against $CCl_4$-induced hepatotoxicity in primary cultures of adult rat hepatocytes. When the extract of PT was directly added into the culture medium at 1, 2, and 5 mg/ml, the extracts not only reduce the $CCl_4$-induced elevation of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase, and lipid peroxide, but also protect cultured hepatocytes from $CCl_4$-induced reduction of reduced glutathione, glutathione reductase, glutathione-S-transferase, glutathione peroxidase, catalase and superoxide dismutase. In addition, the effects of PT water extracts on cytochrome P450 enzymes were relatively marginal, indicating that the hepatoprotective effects of PT extract against $CCl_4$-induced toxicity might not be due to the inhibition of $CCl_4$ activation. In conclusion, the PT extracts were effective in protecting against $CCl_4$ induced hepatotoxicity in hepatocyte cultures, at least in part, by scavenging free radicals, and by modulating enzyme systems involved in cellular oxidative stress.

• Applied Biological Chemistry
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• 제49권3호
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• pp.233-237
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• 2006

Sprague-Dawley 랫트(실험용 쥐)에서 간을 적출하여 Collagen coating된 세포 배양 접시에 간세포 배양을 하여 친환경 재배 배 추출물과 일반 관행 재배 배 추출물의 간세포 기능성을 분석하였다. 3일간 적응 시킨 후 관행 재배 배 추출물과 친환경 재배 배 추출물을 농도 별(동결 건조된 $5\;{\mu}g/ml$, $20\;{\mu}g/ml$, $40\;{\mu}g/ml$) 처리하여 간세포의 에너지원인 ATP assay를 실시한 결과 관행 재배 배 추출물의 경우는 대조군과 유의한 차이는 인정이 되지 않았으나 친환경 재배 배 추출물의 경우 농도 의존적으로 정상군에 비해 180%까지 증가하는 것으로 나타났다. 한편 세포 성장의 경우도 DNA의 특유 염기 구성성분중의 하나인 thymine의 전구체인 $[^3H]$-thymidine을 간세포에 처리하여 DNA 합성을 알아본 결과 친환경 재배 배 추출물 을 농도별로 처리하였을 때 관행재배 배 추출물에 비해 간세포의 세포 성장율이 증가하는 것으로 나타났다($40\;{\mu}g/ml$ 투여 시 관행 재배 배추출물 112% vs. 친환경 재배 배 추출물은 234%; p<0.05). 아울러 세포성장 단백질인 CDK-2, CDK-4의 경우도 친환경 배 추출물 처리 시 현저하게 증가하는 것으로 나타났으며 세포성장 억제 단백질인 p21WAF1/Cip1 및 p27 Kip1의 경우는 억제시키는 것으로 나타났다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.305-306
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• 2003

To treat fulminant hepatic failure (FHF) patients, various extracorporeal bioartificial liver (BAL) systems have been developed. Several requirements should be met for the development of BAL systems: hepatocytes should be cultured in a sufficiently high density; their metabolic functions should be of a sufficiently high level and duration; and the BAL systems module should permit scaling-up and aseptic handling. Several investigators have found that freshly isolated primary hepatocytes can be cultured into three dimensional, tightly packed, freely suspended, multicellular aggregates, or spheroids. These specialized cell structures exhibited enhanced liver specific functions and a prolonged differentiated state compared to cells maintained in a monolayer culture. Cells in spheroids appear to mimic the morphology and ultrastructure of the in vivo liver lobule. The ability of hepatocytes to organize into three-dimensional structures was hypothesized to contribute to their enhanced liver-specific activities. In this study, the ammonia removal rate and urea secretion rate of pig hepatocytes spheroids encapsulated in Ca-alginate bead were determined. A packed-bed bioreactor with encapsulated pig hepatocytes was devised as BAL support system. The efficacy of the system was evaluated in vitro.

• 한국환경과학회:학술대회논문집
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• 한국환경과학회 2003년도 International Symposium on Clean Environment
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• pp.159-164
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• 2003

Effects of Al on vitellogenin (VTG) and VTG mRNA induction by estradiol-17 $\beta$($E_2$) were examined in primary hepatocyte culture of rainbow trout. Hepatocytes were precultured for 2 days and then E2 ($2{\times}10^{-6}$M) and Al ($10^{-6}-10^{-4}$M) were simultaneously added to the incubation medium. Hepatocytes were cultured for 5 more days. Media and hepatocytes were then analyzed by SDS-PAGE and Northern blotting for VTG and VTG mRNA, respectively. These metal had no appreciable effect on the viability of hepatocytes in culture. However, Al interfered with VTG production and VTG mRNA expression. Al reduced VTG production in a concentration-dependent way and a significant reduction accurred at Al concentrations greater than $5{\times}10^{-5}$M. VTG mRNA expression also decreased with a negative correlation with Al concentration (r＝-0.98). These results suggest that Al inhibit VTG production at the transcriptional level to reduce VTG mRNA expression.

• 생약학회지
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• 제36권3호통권142호
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• pp.224-228
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• 2005

Protective effects of various natural flavonoids on carbon tetrachloride $(CCl_4)-induced$ hepatotoxicity were investigated in primary cultured rat hepatocytes. Some of these flavonoids decreased the ALT and LDH releases induced by $CCl_4$ in A dose-dependent manner. Neohesperidin, hesperetin, baicalin, baicalein and quercetin inhibited $CCl_4-induced$ alanine aminotransferase (ALT) release. In addition, quercetin, quercitrin, neohesperidin, baicalin, baicalein and naringin reduced $CCl_4$ induced lactate dehydrogenase (LDH) leakage. Among these flavonoids, quercitrin, quercetin, baicalin and baicalein possessed potent protective effects and were selected for the further investigation on lipid peroxidation. These four flavonoids inhibited dose dependently $CCl_4-induced$ lipid peroxidation. Especially, the protective effects of quercetin and baicalein were similar to silybin as a well-known hepatoprotective agent. These results suggest that these four flavonoids have significant cytoprotective effects and possibility of therapeutic effect on chemical-induced liver diseases.

• 한국약용작물학회지
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• 제8권2호
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• pp.157-164
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• 2000

활성산소에 기인하는 다양한 성인병을 예방 치료할 수 있는 안전하고 우수한 항산화 물질을 천연물로 부터 개발하기 연구의 일환으로 오배자의 주성분에 대한 항산화효과 및 간보호효과를 연구하여 다음과 같은 결과를 얻었다. 1. Ferric-Thiocyanate 법에 의하여 과산화반응 억제에 따른 항산화 활성을 측정한 결과 모두 tocopherol보다 강한 과산화반응 억제 활성을 보였으며 gallic acid methyl ester, protocatechuic acid와 syringic acid는 tocopherol 보다는 강한 활성을 보였으며 BHA와는 유사 또는 그 이상의 활성을 보였다. 특히 1, 2, 3, 4, $6-penta-O-galloyl-{\beta}-D- glucose$는 tocopherol과 BHA보다 매우 강한 과산 화반응억제 활성을 보였다. 2. TBA 법에 의한 과산화지질의 생성의 지표인 MDA 억제효과에 의한 항산화효과를 측정한 결과 gallic acid 는 $1.6{\times}10^{-4}$의 농도에서 BHA 보다 높은 억제율을, protocatechuic acid는 $1.6{\times}10^{-6}$의 농도에서 BHA 보다 높은 억제율을 보였다. gallic acid methyl ester와 1, 2, 3, 4, $6-penta-O-galloyl-{\beta}-D-glucose$는 농도가 증가함에 따라 우수한 MDA의 억제활성을 나타내었다. 3. Glycyrrhizin을 비교군으로 하여 간 보호 실험을 한 결과 1, 2, 3, 4, $6-penta-O-galloyl-{\beta}-D-glucose$의 0.5mg/ml첨가군에서는 GOT의 강한 감소 효과를 보였으며 GPT 실험에서는 각 시료 0.5mg/ml 첨가군에서 감소하는 경향을 나타내었다.