• 약학회지
• /
• 제42권5호
• /
• pp.473-479
• /
• 1998

Lovastatin (LOVA), a fungal metabolite isolated from cultures of Aspergillus terreus, is a competitive HMG-CoA reductase inhibitor used for the treatment of primary hyper cholesterolemia, and has also been shown to suppress growth in a variety of non-glioma tumor cell lines. A sensitive reversed-phase high-perfonnance liquid chromatographic method with ultraviolet (UV) absorbance detection has been developed to quantitate LOVA in human plasma and urine samples using liquid-liquid extraction procedure. Baseline separation of LOVA and internal standard, simvastatin was achieved on a Novapak $C_{18}$ analytical column with a mobile phase containing 0.025M $NaH_2PO_4$: CAN (35:65, v/v%), adjusted pH to 4.5. The flow rate was set at 1.5ml/min, and the column effluent was monitored by a UV detection at 238nm. The limit of quantification was determined to be 0.5${\mu}$g/ml while extraction efficiency of LOVA ranged from 73.4-82.9% at LOVA concentrations of 0.5 to 10${\mu}$g/ml. Good linearity with correlation coefficients greater than 0.999 was obtained in the range of LOVA concentrations from 0.5 to 10${\mu}$g/ml. The accuracy and the precision were proven excellent with relative standard deviation (RSD, %) and relative error (RE, %) of less than 4.2 and 4.0, respectively. Intraday precision, evaluated at five LOVA concentrations (0.5, 1, 2, 5, 10${\mu}$g/ml) and expressed as RSD ranged from 0-1.82% while the interday precision at the same concentrations ranged from 0.7-10.5%. The analytical method described was then successfully employed for the determination of LOVA concentrations in plasma samples obtained during a phase II clinical trial using high doses of LOVA (30-40mg/kg/day). This method could be further utilized for the ongoing pharmacolkinetic studies and therapeutic drug monitoring of the high-dose LOVA therapy in adenocarcinoma patients.

• 대한한방내과학회지
• /
• 제29권2호
• /
• pp.432-442
• /
• 2008

Objective : Ssanggang-tang(SGT) is noted as an effective method to treat hyperlipidemia in China. The main purpose of this study was analysis of the effects of Ssanggang-tang(SGT) on rats induced with caused hyperlipidemia. Methods : We observed 3 experimental groups: normal, control, and SGT. Rats were provided a normal diet in the normal group, and the other groups were provided a hyperlipidemic diet to induce hyperlipidemia. After 2 weeks, SGT was treated in the SGT group. For 6 weeks, values related to hyperlipidemia were observed in the 3 experimental groups. Results : SGT decreased some values related to hyperlipidemia like total cholesterol, low density lipoprotein cholesterol(LDL cholesterol), triglyceride, acetocoenzyme A acetyltransferase(ACAT), and 3-hydroxy-3-methylglutaryl coenzyme A reductase(HMG-CoA reductase). However, it showed no effect on weight change or high density lipoprotein cholesterol(HDL cholesterol). Conclusion : These results suggest that SGT might be effective in treatment and prevention of hyperlipidemia.

• Journal of Microbiology and Biotechnology
• /
• 제26권2호
• /
• pp.295-308
• /
• 2016

This study investigated the effects of a flavonoid-rich ethanol extract of persimmon leaf (PL), an ethanol extract of Citrus junos Sieb (CJS), and a PL-CJS mixture (MPC) on mice fed a high-fat diet (HFD). We sought to elucidate the mechanisms of biological activity of these substances using measurements of blood coagulation indices and lipid metabolism parameters. C57BL/6J mice were fed a HFD with PL (0.5% (w/w)), CJS (0.1% (w/w)), or MPC (PL 0.5%, CJS 0.1% (w/w)) for 10 weeks. In comparison with data obtained for mice in the untreated HFD group, consumption of MPC remarkably prolonged the activated partial thromboplastin time (aPTT) and prothrombin time (PT), whereas exposure to PL prolonged aPTT only. Lower levels of plasma total cholesterol, hepatic cholesterol, and erythrocyte thiobarbituric acid-reactive substances, hepatic HMG-CoA reductase, and decreased SREBP-1c gene expression were observed in mice that received PL and MPC supplements compared with the respective values detected in the untreated HFD animals. Our results indicate that PL and MPC may have beneficial effects on blood circulation and lipid metabolism in obese mice.

• Journal of Microbiology and Biotechnology
• /
• 제21권7호
• /
• pp.766-775
• /
• 2011

We investigated the effect of high molecular weight polygamma- glutamic acid (hm ${\gamma}$-PGA) on adiposity and lipid metabolism of rats in the presence of an obesity-inducing diet. Thirty-two Sprague-Dawley rats were fed either a normal-fat (11.4% kcal fat, NFC) or high-fat (51% kcal fat, HFC) diet. After 5 weeks, half of each diet-fed group was treated with hm ${\gamma}$-PGA (NFP or HFP) for 4 weeks. The HFC group had significantly higher body weight, visceral fat mass, fasting serum levels of total cholesterol, LDL cholesterol, and leptin, and lower serum HDL cholesterol level compared with those of the NFC group (p < 0.05). Treatment with hm ${\gamma}$-PGA decreased body weight gain and perirenal fat mass (p<0.05), fasting serum total cholesterol, and mRNA expression of glucose-6- phosphate dehydrogenase (G6PD), regardless of dietary fat contents (p < 0.01). However, hm ${\gamma}$-PGA increased serum HDL cholesterol in the HFC group (p < 0.05). In vitro, 3-hydroxy-3-methylglutaryl coenzyme-A (HMGCoA) reductase activity was suppressed by the addition of hm ${\gamma}$-PGA. In agreement with observations in animal study, the supplementation of hm ${\gamma}$-PGA (150 mg/day) to 20 female subjects in an 8-week double-blind, placebocontrolled study resulted in a tendency to decrease total cholesterol and LDL cholesterol concentrations. We thus conclude that dietary supplementation of hm ${\gamma}$-PGA may act as a hypocholestrolemic agent, secondary to its inhibitor effect on HMG-CoA reductase, and decrease abdominal adiposity by decreasing hepatic lipogenesis. The present study is an important first step in establishing the effect of hm ${\gamma}$-PGA on cholesterol levels in rats and humans.

• Plant Biotechnology Reports
• /
• 제5권1호
• /
• pp.53-60
• /
• 2011

Artemisinin is effective against both chloroquine-resistant and -sensitive strains of Plasmodium species. However, the low yield of artemisinin from cultivated and wild plants is a serious limitation to the commercialization of this drug. Optimization of artemisinin yield either in vivo or in vitro is therefore highly desirable. To this end, we have overexpressed the 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGR) gene (hmgr) from Catharanthus roseus L. in Artemisia annua L. and analyzed its influence on artemisinin content. PCR and Southern blot analyses revealed that the transgenic plants showed stable integration of the foreign hmgr gene. The reverse transcriptase-PCR results suggested that the hmgr was expressed at the transcriptional level in transgenic lines of Artemisia annua L., while the high-performance liquid chromatography analysis showed that artemisinin content was significantly increased in a number of the transgenic lines. Artemisinin content in one of the A. annua transgenic lines was 38.9% higher than that in non-transgenic plants, and HMGR enzyme activity in transgenic A. annua L. was also higher than that in the non-transgenic lines.

• 약학회지
• /
• 제57권1호
• /
• pp.24-31
• /
• 2013

Although statins, inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, have been shown to increase melanin synthesis, the exact mechanism of this action is not fully understood. In this study we investigated the possible involvement of intracellular $Ca^{2+}$ signal in the mechanism of stimulation of melanin synthesis induced by lovastatin in B16 cells. Lovastatin stimulated the production of melanin in a dose-dependent manner in the cells. Treatment with mevalonate, FPP and GGPP, precursors of cholesterol, did not significantly suppress the lovastatin-induced melanin production, suggesting that inhibition of cholesterol synthesis may not be involved in the mechanism of the action of lovastatin. In addition, lovastatin did not significantly alter the cAMP concentration and the stimulated production of melanin by lovastatin was not significantly changed by treatment with H89, a potent inhibitor of protein kinase A, which demonstrates that cAMP pathway may not be involved. However, lovastatin increased intracellular $Ca^{2+}$ concentration in a dose-related fashion. Treatment with EGTA, an extracellular $Ca^{2+}$ chelator did not significantly alter the lovastatin-induced intracellular $Ca^{2+}$ increase and melanin synthesis, whereas intracellular $Ca^{2+}$ reduction with BAPTA/AM and intracellular $Ca^{2+}$ release blockers (dantrolene and TMB-8) completely blunted these actions of lovastatin. Taken together, these results suggest that the intracellular $Ca^{2+}$ release may play an important role in the lovastatin-induced stimulation of melanin synthesis in B16 cells. These results further suggest that lovastatin may be useful for the treatment of hypopigmentation disorders, such as vitiligo.

• 대한한의학방제학회지
• /
• 제22권1호
• /
• pp.93-104
• /
• 2014

Objectives : Samhwangsasim-tang (SHSST) is a traditional Korean medication, which has been used in Korea for treatment of hypertension and chest pain. Hyperlipidemia and inflammation could influence hypertension and chest pain. This study investigated whether and how SHSST reduces the hyperlipidemia and inflammation related to high-cholesterol diet-induced obesity in rats. Methods : Mice were divided randomly into four groups: the normal diet group, high-cholesterol diet group, low dose treatment group supplemented with 30% ethanol extract of SHSST (L) and high dose treatment group supplemented with 80% ethanol extract of SHSST (H). L and H groups were orally administered with SHSST at the dose of 50mg/kg a day respectively and others were administered with the same volume of physiological saline. Results : Administration of SHSST resulted in a decrease in serum levels of total cholesterol and low-density lipoprotein. Expression of hepatic genes(SREBP2, LXR, LDLR, and HMG-CoA) related with cholesterol metabolism was also suppressed. In addition, SHSST decreased the expression of inflammation-related gene (TNF-${\alpha}$, IL-6, ICAM-1, VCAM-1, TGF-${\beta}1$ and fibronectin). Histological examinations also showed that the size of the adipocytes was smaller in the SHSST treated group than in the high-colesterol diet group. In an in vitro study, SHSST inhibited the production of nitric oxide in a concentration-dependent manner. Conclusions : This study indicates that SHSST has anti-hyperlipidemia and anti-inflammatory effects. It may also suggest that SHSST may be alternative therapy for treatment of hyperlipidemia and its complications.

• 한국식품영양학회지
• /
• 제19권2호
• /
• pp.176-182
• /
• 2006

Sorghum bicolor L. Moench(Sorghum, Su-Su) is a major cereal food crop used in many parts of the world. It is used as a human food resource and folk medicines in Asia and Africa. The stem of sorghum has been used as a digestive aid and an anti-diarrheal agent. Sorghum hybrids contain high levels of diverse phenolic compounds that may provide health benefits. High levels of polyflavanols, anthocyanins, phenolic acids, and other antioxidant compounds have been reported in sorghums, which have also been shown to possess various biological activities such as anti-mutagenic, anti-carcinogenic, and HMG-CoA reductase inhibitory activities. In an in vitro experiment, we examined mice splenocyte proliferation and production of three types of cytokine($IL-1{\beta},\;IL-6,\;TNF-{\alpha}$) by peritoneal macrophages cultured with ethanol and water extracts of Sorghum bicolor L. Moench. A single cell suspension of splenocytes was prepared and the cell proliferation of the splenocytes was examined by MTT assay. The splenocyte proliferation was increased when water extracts of Sorghum bicolor L. Moench were used as supplements in all concentrations investigated. The production of cytokine($IL-1{\beta},\;IL-6,\;TNF-{\alpha}$) by activated peritoneal macrophage was detected by ELISA using the cytokine kit. $IL-1{\beta},\;IL-6,\;and\;TNF-{\alpha}$ production by activated macrophages were increased by supplementation with Sorghum bicolor L. Moench water extracts. This study suggests that supplementation of with Sorghum bicolor L. Moench water extracts may enhance immune function by regulating the splenocyte proliferation and enhancing the cytokine production by activated macrophages in vitro.

• Clinical and Experimental Reproductive Medicine
• /
• 제26권1호
• /
• pp.75-81
• /
• 1999

The beneficial effect of glucose and phosphate ions in culture medium on the development of human embryos in vitro has not been fully elucidated. The purpose of this study was to evaluate the influence of fertilization and culture of embryos in glucose/phosphate-free m-TALP medium on pregnancy rates in IVF-ET program. The patients in 244 IVF-ET cycles received GnRH agonist + HMG regimens. A does of 10,000 IU HCG was administered when two or more dominent follicles reached 18mm in diameter. Thirty-six hours after HCG, oocytes were recovered transvaginally using ultrasound guidance. Aspirated oocytes were matured for 4 to 6 h in TCM-199 supplemented with 10% follicular fluid (FF). Insemination was carried out with 50,000 motile spermatozoa in TCM-199 + 10% FF or m-TALP + 5% FF + 5% fetal cord serum (FCS) according to experimental design. After 6 h, oocytes were washed 3 to 4 times and cultured in each fresh medium. After 20 h, oocytes were freed from cumulus/corona cells and examined for the presence of pronuclei. Fertilized oocytes were transferred into each co-culture drops and cultured for further incubation. On day 3, embryo transfer was performed with grade 1 and 2 embryos. Monolayers for co-culture of embryos were prepared by plating $1{\times}10^5$ cumulus cells/ml in 10ul drop of TCM-199 + 10% FF or m-TALP + 5% FF + 5% FCS media 24 h prior to the onset of co-culture. Development to 4 to 16 cell stage was observed at 70x magnification following two days of incubation. Pregnancy was confirmed by detecting increasing serum ${\beta}$-hCG concentrations for 11 days following embryo transfer. Data were analyzed by ${\chi}^2$-test. Oocytes from 244 IVF-ET cycles were randomized. The number of cycles and mean age of patients were 97 and 147, 31.3 yrs and 31.2 yrs for TCM-199 (control) and m-TALP groups, respectively. The mean number of retrieved oocytes/cycle, fertilization rates, number of embryos transferred/ET and pregnancy rates were 11.1 and 10.3, 65.1% and 67.3%, 4.1 and 4.7, 28.9% and 43.8% for TCM-199 and m-TALP groups, respectively. Differences in the pregnancy rates were found between control and m-TALP groups (p<0.05). The pregnancy rate of patients divided according to maternal age groups of ${\leq}30$, 31-35, $36{\leq}$ were 44.4% and 49.0%, 26.1% and 41.3%, 29.2% and 41.2% for control and m-TALP groups, respectively. These data indicate that culture of human embryos in glucose/phosphate-free m-TALP medium improves pregnancy rates.

• 한국식품위생안전성학회지
• /
• 제28권2호
• /
• pp.89-94
• /
• 2013

마늘 숙성 중 생성되는 S-allylmercaptocysteine의 콜레스테롤 생합성 억제 효과에 대하여 Hepatocytes를 이용하여 조사하였다. HepG2 cells을 Dulbecco's modified Eagle's medium (DMEM)에 배양하여 S-allylmercaptocysteine의 농도를 20, 40, 60, 80 및 100 mL 씩 각각 첨가하여 cell viability를 살펴본 결과 20~40 ${\mu}g/mL$에서는 높았으며, 60 ${\mu}g/mL$ 농도에서 약 50%가 유지되었다. S-allylmercaptocysteine을 5, 10, 15 및 20 ${\mu}g/mL$ 농도로 [$^{14}C$]-acetatecholesterol에서 처리하였을 경우 15 ${\mu}g/mL$ 농도에서 cholesterol 생합성이 79%로 억제되었다. Fatty acid synthase의 활성은 0.95 nmol에서 19%의 억제효과를 나타내었으나, Glucose 6-phosphate dehydrogenase (G6PDH)의 활성에는 거의 영향을 미치지 않았다. S-allylmercaptocysteine의 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase의 활성은 용량 의존형으로 감소하였다. 이상의 결과로 보아 마늘 숙성 과정에서 생성되는 주요 성분인 S-allylmercaptocysteine은 간 세포에서 cholesterol의 생합성을 억제하는데 기여하는 것으로 나타났다.