Cloning and functional verification of carboxyl CoA ligases(AeCCLs) in Arnebia euchroma / 中国中药杂志

Carboxyl CoA ligases(CCLs) is an important branch of adenylate synthetase gene family, which mainly has two-step catalytic reactions. Firstly, in the presence of adenosine triphosphate, it can catalyze the pyrophosphorylation of carboxylateswith diffe-rent structures to form corresponding acyl adenosine monophosphate intermediates. Secondly, adenosine monophosphate was replaced by free electrons in the mercaptan group of enzyme A or other acyl receptors by nucleophilic attack to form thioesters. In this study, on the basis of the transcriptome database of Arnebia euchroma, two genes were selected, named AeCCL5(XP_019237476.1) and AeCCL7(XP_019237476.1). Bioinformatics analysis showed that their relative molecular weights were 60.569 kDa and 60.928 kDa, theoretical PI were 8.59 and 8.92, respectively. They both have transmembrane domains but without signal peptide. By multiple sequence alignment and phylogenetic tree analysis, we found that the similarity between AeCCLs and other plant homologous proteins was not high, and the substrate binding sites of AeCCLs were not highly conserved. The reasons might be that the sequence and structure need to adapt to the changes of new substrates in the process of evolution. In this study, the full-length of AeCCL5 and AecCCL7 were cloned into the expression vector pCDFDuet-1. The proteins of AeCCL5 and AeCCL7 with His-tag were expressed in Escherichia coli. The proteins of AeCCL5 and AeCCL7 were purified by nickel column. In vitro enzymatic reactions proved that both AeCCL5 and AeCCL7 can participate in the upstream phenylpropane pathway of shikonin biosynthesisby catalyzing 4-coumaric acid to produce 4-coumarin-CoA, and then to synthesis p-hydroxybenzoic acid, which is an important precursor of shikonin biosynthesis in A. euchroma.

Cloning and prokaryotic expression of 3-ketoacyl-CoA thiolase gene AIKAT from Atractylodes lancea / 中国中药杂志

In this study, the gene encoding the key enzyme 3-ketoacyl-CoA thiolase(KAT) in the fatty acid β-oxidation pathway of Atractylodes lancea was cloned. Meanwhile, bioinformatics analysis, prokaryotic expression and gene expression analysis were carried out, which laid a foundation for the study of fatty acid β-oxidation mechanism of A. lancea. The full-length sequence of the gene was cloned by RT-PCR with the specific primers designed according to the sequence information of KAT gene in the transcriptomic data of A. lancea and designated as AIKAT(GenBank accession number MW665111). The results showed that the open reading frame(ORF) of AIKAT was 1 323 bp, encoding 440 amino acid. The deduced protein had a theoretical molecular weight of 46 344.36 and an isoelectric point of 8.92. AIKAT was predicted to be a stable alkaline protein without transmembrane segment. The secondary structure of AIKAT was predicted to be mainly composed of α-helix. The tertiary structure of AIKAT protein was predicted by homology modeling method. Homologous alignment revealed that AIKAT shared high sequence identity with the KAT proteins(AaKAT2, CcKAT2, RgKAT and AtKAT, respectively) of Artemisia annua, Cynara cardunculus var. scolymus, Rehmannia glutinosa and Arabidopsis thaliana. The phylogenetic analysis showed that AIKAT clustered with CcKAT2, confirming the homology of 3-ketoacyl-CoA thiolase genes in Compositae. The prokaryotic expression vector pET-32 a-AIKAT was constructed and transformed into Escherichia coli BL21(DE3) for protein expression. The target protein was successfully expressed as a soluble protein of about 64 kDa. A real-time quantitative PCR analysis was performed to profile the AIKAT expression in different tissues of A. lancea. The results demonstrated that the expression level of AIKAT was the highest in rhizome, followed by that in leaves and stems. In this study, the full-length cDNA of AIKAT was cloned and expressed in E. coli BL21(DE3), and qRT-PCR showed the differential expression of this gene in different tissues, which laid a foundation for further research on the molecular mechanism of fatty acid β-oxidation in A. lancea.

Detection of some virulence factors genes in Staphylococcus aureus isolated from different clinical cases

Staphylococcus aureus is one of the most important pathogenic type to humans, and the most common species responsible for a wide range of diseases such as furuncles, various abscesses, wounds abscesses resulting from surgical operations, dermatitis, soft tissue inflammation, arthritis, bones inflammation, bronchial pneumonia, inflammation of internal parts of the heart and injuries caused by toxins such as toxic shock syndorome and staphylococcus aureus syndrome and food poisoning. The current study aimed by finding the genes responsible for the virulence factors in S. aureus isolates by using the Single and Multiplex PCR mechanism (technology). A total of 60 specimens (urine, burn swabs, wound swaabs) from different clinical cases were collected from patients (in different age groups) who admitted to several health centers in Al-Diwaniyah Teaching Hospital, Iraq, during a period extending from October 2016 to January 2017. Some virulence factors were investigated for 30 isolate only of MRSA using Single and Multiplex PCR for detection virulence factor genes which both coa gene encoding production of coagulase, clfA gene encoding for clumpting factor, spa gene encoding for protein A, fnbA gene encoding for fibronectin binding proteins, luks gene encoding prouction of Panton Valentine Leukocidin (PVL). Results 30 (100%) were possess coa, clfA, spa and fnbA genes, 13 (43.3%) were possess luks gene

Nicorandil alleviated cardiac hypoxia/reoxygenation-induced cytotoxicity via upregulating ketone body metabolism and ACAT1 activity

To study the effect of nicorandil pretreatment on ketone body metabolism and Acetyl-CoA acetyltransferase (ACAT1) activity in hypoxia/reoxygenation (H/R)-induced cardiomyocytes. In our study, we applied H9c2 cardiomyocytes cell line to evaluate the cardioprotective effects of nicorandil. We detected mitochondrial viability, cellular apoptosis, reactive oxygen species (ROS) production and calcium overloading in H9c2 cells that exposed to H/R-induced cytotoxicity. Then we evaluated whether nicorandil possibly regulated ketone body, mainly β-hydroxybutyrate (BHB) and acetoacetate (ACAC), metabolism by regulating ACAT1 and Succinyl-CoA:3-keto-acid coenzyme A transferase 1 (OXCT1) protein and gene expressions. Nicorandil protected H9c2 cardiomyocytes against H/R-induced cytotoxicity dose-dependently by mitochondria-mediated anti-apoptosis pathway. Nicorandil significantly decreased cellular apoptotic rate and enhanced the ratio of Bcl-2/Bax expressions. Further, nicorandil decreased the production of ROS and alleviated calcium overloading in H/R-induced H9c2 cells. In crucial, nicorandil upregulated ACAT1 and OXCT1 protein expressions and either of their gene expressions, contributing to increased production of cellular BHB and ACAC. Nicorandil alleviated cardiomyocytes H/R-induced cytotoxicity through upregulating ACAT1/OXCT1 activity and ketone body metabolism, which might be a potential mechanism for emerging study of nicorandil and other K(ATP) channel openers.

Comparison of Adverse Events between High-intensity and Moderate- to Low-intensity Statin Group

BACKGROUND: 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) effectively reduce serum levels of low-density lipoprotein (LDL) and total cholesterol. High-intensity statins are recommended for all patients aged ≤75 with clinical atherosclerotic cardiovascular disease (ASCVD), diabetes mellitus aged 40–75 with ≥7.5% estimated 10-year ASCVD risk and LDL-C ≥190 mg/ dL. High-intensity statins associated with more frequent adverse events (AEs) compared to moderate- to low-intensity statins. The aim of this study was to compare AEs between high-intensity and moderate- to low-intensity statin group using the Korea Adverse Event Reporting System (KAERS) database. METHODS: Adults (≥18 years) with statin-associated AEs from July 2009-June 2014 were included. Only AEs classified as “certain”, “probable” and “possible” based on the WHO-Uppsala Monitoring Center criteria were analyzed. RESULTS: In total, 247 AEs from 196 patients [high-intensity statin group (HG), n = 25 (13%); moderate- to lowintensity statin group (MLG), n = 171 (87%)] were included. Mean age was higher in HG compared with MLG (67 ± 14 vs 62 ± 12). The HG showed a significant higher frequency of liver/biliary system disorders (37% vs 14%, p = 0.001). Hepatic function abnormal was reported more frequently in HG compared to MLG (26% vs 9%, p = 0.006). CONCLUSION: According to KAERS data, liver/biliary system disorders were more frequently reported in HG compared to MLG.

The Risk of Rosacea According to Chronic Diseases and Medications: A 5-Year Retrospective, Multi-Institutional Case-Control Study

BACKGROUND: Rosacea is associated with chronic systemic disease. However, research is lacking in Asian countries. OBJECTIVE: To evaluate the association between rosacea and cardiovascular diseases (CVDs) related systemic comorbidities, and the use of antihypertensive and antihyperlipidemic drugs in Korea. METHODS: A five-year retrospective study, using hospital database, was conducted in five medical centers for five years. Totally 1,399,528 patients were evaluated. RESULTS: The overall frequency for diagnosed rosacea was 0.18% over five years (2,536 rosacea patients). Patients with diabetes and patients with dyslipidemia were more likely to have rosacea (odd ratio [OR] 2.724, 95% confidence interval [CI] 1.295~5.730, p=0.016; OR 1.788, 95% CI 1.445~2.212, p < 0.001). Patients with CVD were less likely to have rosacea (OR 0.431, 95% CI 0.244~0.760, p=0.003). Patients with α-blocker prescriptions and patients with β-blocker prescriptions showed a tendency diagnosed with rosacea frequently (OR 1.963, 95% CI 1.200~3.212, p=0.006; OR 3.939, 95% CI 3.512~4.419, p < 0.001). Patients with [beta]-hydroxy-[beta]-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, and those with fibrate, were prone to have rosacea (OR 1.599, 95% CI 1.390~1.839, p < 0.001; OR 1.660, 95% CI 1.056~2.609, p=0.026). As adjusted results, among the patients who took HMG-CoA reductase inhibitor without dyslipidemia, rosacea was less likely to be diagnosed (OR 0.780, 95% CI 0.620~0.982, p=0.034). CONCLUSION: Rosacea is associated with chronic diseases and drugs.

Candidatos a novos agentes tuberculostáticos: planejamento e sí­ntese de inibidores da fosfopanteteí­na adenililtransferase / New antituberculosis agents' candidates: design and synthesis of phosphopantetheine adenylyltransferase inhibitors

A tuberculose (TB) é uma das maiores causas de morte por infecção no mundo, sendo que, em 2015, registraram-se 10,4 milhões de novos casos. O agente etiológico da doença, o Mycobacterium tuberculosis (Mtb), apresenta altos níveis de resistência frente aos quimioterápicos disponíveis para o tratamento da TB. Além disso, a terapia atual da doença explora poucos alvos essenciais ao Mtb. Neste sentido, explorar novos alvos, essenciais ao crescimento e sobrevivência da micobactéria é de grande interesse e poderia gerar fármacos mais efetivos, eficazes contra cepas resistentes e a forma latente da TB. Para este fim, o presente trabalho propôs o desenvolvimento de inibidores da enzima fosfopanteteína adenililtransferase (PPAT), a qual possui caráter regulatório na via de biossíntese da Coenzima A (CoA) da micobactéria. Inicialmente, propuseram-se 50 estruturas de potenciais inibidores da PPAT de M. tuberculosis (MtPPAT), baseando-se na estrutura de seu substrato, a fosfopanteteína, e na estrutura do sítio ativo da enzima. Em seguida, propuseram-se outros 28 ligantes. A fim de se prever as potenciais complementaridades entre os 78 inibidores propostos e o sítio ativo da MtPPAT, empregou-se a estratégia de docking. Posteriormente, realizaram-se cálculos semi-empíricos, com os complexos dos ligantes que se mostraram mais interessantes nas simulações de docking, a fim de se obter informações sobre a entalpia de interação dos ligantes com o sítio ativo da MtPPAT. A partir dos resultados obtidos nos estudos computacionais, selecionaram-se os inibidores que se mostraram mais promissores. A síntese destes ligantes e a de seus fragmentos foi realizada. Avaliaram-se a atividade microbiológica in vitro, bem como a citotoxicidade dos ligantes sintetizados. Alguns dos compostos sintetizados apresentaram atividade frente às cepas sensíveis e resistentes do Mtb na casa de micromolar. Todos os compostos ativos não foram considerados citotóxicos. A fim de validar o planejamento e o alvo dos possíveis inibidores, verificando a atividade inibitória desses frente à enzima MtPPAT, realizou-se a produção e purificação da enzima. Por fim, realizaram-se ensaios de inibição enzimática frente à MtPPAT, os quais permitiram a identificação dos primeiros inibidores da enzima já descritos, com atividade na casa de micromolar, validando-se o alvo em questão

Tuberculosis is one of the major causes of death by infection worldwide. In 2015, 10.4 thousand new cases of the disease were registered. The tuberculosis' causing agent Mycobacterium tuberculosis presents high levels of resistance for the available chemotherapy. Thereof, exploit new M. tuberculosis targets is of utmost importance to overcome drug resistant tuberculosis. In this sense, the enzyme phosphopantetheine adenylyltransferase (PPAT) generates scientific interest since it displays a regulatory role in the M. tuberculosis coenzyme A (CoA) biosynthesis. Therefore, the purpose of the present study was the development of M. tuberculosis PPAT (MtPPAT) inhibitors. Initially, 50 potentially MtPPAT inhibitors were designed based on MtPPAT's substrate and the enzyme's active site. After preliminary results, more 28 compounds were designed. Docking simulations were performed with the 78 compounds synthesized, leading to the prediction of the interaction between the proposed inhibitors and MtPPAT active site. Latelly, semi-empirical calculations were performed with the most promising compounds. These calculations were carried out to obtain information about the enthalpy interactions between compounds and MtPPAT active site. Computational studies led to the selection of the most promising inhibitors. Those compounds and some of their fragments were synthesized, purified, and characterized. The synthesized compounds had their in vitro microbiological activity and cytotoxicity evaluated. Some of the synthesized compounds showed activity against the Mtb sensitive and resistant strains in micromolar range. Besides that, the active compounds were not considered cytotoxic. To validate the potential inhibitors' design and evaluate their capacity to inhibit MtPPAT, the enzyme was produced and purified. MtPPAT inhibitory assays were performed, leading to the first inhibitors of the enzyme, with activity in micromolar range, validating the target

Chemoprevention of Gastric Cancer: Statins

Gastric cancer is one of the leading causes of cancer-related deaths worldwide. Because the gastric cancer-related mortality rate is expected to increase, chemo-preventive strategies for gastric cancer are required. Statins inhibit 3-hydroxy-3-methylglutaryl coenzyme A reductase, the enzyme involved in the rate-limiting step in cholesterol synthesis. Several studies have shown that statins have potential protective effects against gastric cancer; however, other studies have reported contradictory results. Therefore, further studies are needed to clarify the role of statins in the chemoprevention of gastric cancer.

Comparative pharmacokinetic and tolerability evaluation of two simvastatin 20 mg formulations in healthy Korean male volunteers

Simvastatin is used to reduce plasma cholesterol by inhibiting 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase and is primarily used to treat hypercholesterolemia. This study was conducted to assess the bioequivalence between the generic formulation of simvastatin 20 mg and the branded formulation of simvastatin 20 mg. A generic formulation of simvastatin 20 mg tablet was developed and the pharmacokinetics of the generic formulation were compared with those of the branded formulation of simvastatin 20 mg tablet in 33 healthy male volunteers after a single oral dose in a randomized, open-label, two-period, two-sequence, crossover study. The reference (Zocor®, MSD Korea LTD.) and test (Simvarotin®, Korea Arlico Pharm Co., Ltd.) formulations, two 20 mg tablets each, were administered to all subjects in fasting status. The serial blood samples for pharmacokinetic analysis were collected before dosing and up to 24 hours post-dose, and plasma concentrations of simvastatin were determined by liquid chromatography-tandem mass spectrometry. The pharmacokinetic parameters including T(max), C(max), AUC(last), AUC(inf) and t½ were calculated for both formulations by non-compartmental method, and the log-transformed C(max) and AUC(last) were compared statistically. Geometric mean ratios (90% confidence intervals) of the test to the reference formulation in C(max) and AUC(last) were 0.9652 (0.8302–1.1223) and 0.9891 (0.8541–1.1455), respectively. No significant differences in tolerability profiles were noted between the two formulations. The two formulations of simvastatin 20 mg tablets exhibited comparable pharmacokinetic profiles and 90% confidence intervals were within the acceptable range of bioequivalence criteria.

Differential Immunohistochemical Profiles for Distinguishing Prostate Carcinoma and Urothelial Carcinoma

BACKGROUND: The pathologic distinction between high-grade prostate adenocarcinoma (PAC) involving the urinary bladder and high-grade urothelial carcinoma (UC) infiltrating the prostate can be difficult. However, making this distinction is clinically important because of the different treatment modalities for these two entities. METHODS: A total of 249 patient cases (PAC, 111 cases; UC, 138 cases) collected between June 1995 and July 2009 at Seoul St. Mary's Hospital were studied. An immunohistochemical evaluation of prostatic markers (prostate-specific antigen [PSA], prostate-specific membrane antigen [PSMA], prostate acid phosphatase [PAP], P501s, NKX3.1, and α-methylacyl coenzyme A racemase [AMACR]) and urothelial markers (CK34βE12, p63, thrombomodulin, S100P, and GATA binding protein 3 [GATA3]) was performed using tissue microarrays from each tumor. RESULTS: The sensitivities of prostatic markers in PAC were 100% for PSA, 83.8% for PSMA, 91.9% for PAP, 93.7% for P501s, 88.3% for NKX 3.1, and 66.7% for AMACR. However, the urothelial markers CK34βE12, p63, thrombomodulin, S100P, and GATA3 were also positive in 1.8%, 0%, 0%, 3.6%, and 0% of PAC, respectively. The sensitivities of urothelial markers in UC were 75.4% for CK34βE12, 73.9% for p63, 45.7% for thrombomodulin, 22.5% for S100P, and 84.8% for GATA3. Conversely, the prostatic markers PSA, PSMA, PAP, P501s, NKX3.1, and AMACR were also positive in 9.4%, 0.7%, 18.8%, 0.7%, 0%, and 8.7% of UCs, respectively. CONCLUSIONS: Prostatic and urothelial markers, including PSA, NKX3.1, p63, thrombomodulin, and GATA3 are very useful for differentiating PAC from UC. The optimal combination of prostatic and urothelial markers could improve the ability to differentiate PAC from UC pathologically.

The Role of Pyruvate Dehydrogenase Kinase in Diabetes and Obesity

The pyruvate dehydrogenase complex (PDC) is an emerging target for the treatment of metabolic syndrome. To maintain a steady-state concentration of adenosine triphosphate during the feed-fast cycle, cells require efficient utilization of fatty acid and glucose, which is controlled by the PDC. The PDC converts pyruvate, coenzyme A (CoA), and oxidized nicotinamide adenine dinucleotide (NAD+) into acetyl-CoA, reduced form of nicotinamide adenine dinucleotide (NADH), and carbon dioxide. The activity of the PDC is up- and down-regulated by pyruvate dehydrogenase kinase and pyruvate dehydrogenase phosphatase, respectively. In addition, pyruvate is a key intermediate of glucose oxidation and an important precursor for the synthesis of glucose, glycerol, fatty acids, and nonessential amino acids.

Cholesterol Lowering Effects of Low-dose Statins in Korean Patients

OBJECTIVE: The aim of this study is to compare cholesterol lowering effects of low dose 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) in Korean patients. METHODS: A total of 909 consecutive patients were enrolled prospectively according to the criteria of National Cholesterol Education Program guidelines. Lipid profiles were obtained before and 2 months after statin therapy. RESULTS: Atorvastatin 10 mg (n=260), lovastatin 20 mg (n=145), pitavastatin 2 mg (n=80), pravastatin 20 mg (n=28), rosuvastatin 5 mg (n=145), and simvastatin 20 mg (n=208) reduced low density lipoprotein (LDL) cholesterol by -41.8+/-11.0%, -33.8+/-12.8%, -39.3+/-10.8%, -31.5+/-8.9%, -48.8+/-12.3%, and -42.8+/-13.5%, respectively. LDL cholesterol less than 130 mg/dL was achieved in 90.3%, 76.9%, 88.5%, 85.2%, 97.2%, and 94.2%, respectively. The reduction of LDL cholesterol by 30% or more was obtained in 84.4%, 60.7%, 81.6%, 63.0%, 93.0%, and 83.5%, respectively. LDL cholesterol less than 70 mg/dL or the reduction by 50% or more was observed in a small portion of patients and was variable according to the different types of statins. CONCLUSION: A low dose statin was enough to manage dyslipidemia in most Korean patients with low to moderate risks and was even effective in a subpopulation of high risk patients.

Antioxidant Effects of Statins in Patients with Atherosclerotic Cerebrovascular Disease

BACKGROUND AND PURPOSE: Oxidative stress is involved in the pathophysiological mechanisms of stroke (e.g., atherosclerosis) and brain injury after ischemic stroke. Statins, which inhibit 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, have both pleiotropic and low-density lipoprotein (LDL)-lowering properties. Recent trials have shown that high-dose statins reduce the risk of cerebrovascular events. However, there is a paucity of data regarding the changes in the oxidative stress markers in patients with atherosclerotic stroke after statin use. This study evaluated changes in oxidative stress markers after short-term use of a high-dose statin in patients with atherosclerotic stroke. METHODS: Rosuvastatin was administered at a dose of 20 mg/day to 99 patients who had suffered an atherosclerotic stroke and no prior statin use. Blood samples were collected before and 1 month after dosing, and the serum levels of four oxidative stress markers-malondialdehyde (MDA), oxidized LDL (oxLDL), protein carbonyl content (PCO), and 8-hydroxy-2'-deoxyguanosine (8-OHdG)-were evaluated to determine the oxidation of MDA and lipids, proteins, and DNA, respectively, at both of those time points. RESULTS: The baseline levels and the degrees of reduction after statin use differed among the oxidative stress markers measured. MDA and PCO levels were associated with infarct volumes on diffusion-weighted imaging (r=0.551, p<0.05, and r=0.444, p=0.05, respectively). Statin use decreased MDA and oxLDL levels (both p<0.05) but not the PCO or 8-OHdG level. While the reduction in MDA levels after statin use was not associated with changes in cholesterol, that in oxLDL levels was proportional to the reductions in cholesterol (r=0.479, p<0.01), LDL (r=0.459, p<0.01), and apolipoprotein B (r=0.444, p<0.05). CONCLUSIONS: The impact of individual oxidative stress markers differs with time after ischemic stroke, suggesting that different oxidative markers reflect different aspects of oxidative stress. In addition, short-term use of a statin exerts antioxidant effects against lipid peroxidation via lipid-lowering-dependent and -independent mechanisms, but not against protein or DNA oxidation in atherosclerotic stroke patients.

Hypocholesterolemic metabolism of dietary red pericarp glutinous rice rich in phenolic compounds in mice fed a high cholesterol diet

BACKGROUND/OBJECTIVES: The purpose of the current study was to investigate the effect of red pericarp glutinous rice rich in polyphenols (Jakwangchalbyeo, red rice) on serum and hepatic levels of cholesterol and hepatic protein expression linked to synthesis and degradation of cholesterol in a hypercholesterolemic mice diet as compared with brown rice. MATERIALS/METHODS: C57BL/6 male mice were randomly divided into four groups (n = 5 each), which were fed different diets for a period of 12 weeks: American Institute of Nutrition (AIN)-93G diet, AIN-93G diet with 2% cholesterol, brown rice with 2% cholesterol, or red rice with 2% cholesterol. RESULT: Consumption of red rice resulted in a significant decrease in serum level of low-density lipoprotein cholesterol and hepatic levels of triglyceride and total-cholesterol. Expression of acyl-coenzyme A cholesterol acyltransferase-2 (ACAT-2), sterol regulatory element binding protein-2 (SREBP-2), and 3-hydroxyl-3-methylglutaryl coenzyme A (HMG-CoA) reductase was decreased, while expression of phosphorylated adenosine monophosphate activated protein kinase (p-AMPK)/AMPK ratio, cholesterol 7-alpha-hydroxylase (CYP7a1), and sterol 12-alpha-hydroxylase (CYP8b1) was increased in mice fed red rice. Brown rice had similar effects on cholesterol metabolism, but the effect of red rice was significantly greater than that of brown rice. CONCLUSIONS: The current study suggested that red rice had a hypocholesterolemic effect by lowering hepatic cholesterol synthesis through ACAT-2, HMG-CoA reductase, and SREBP-2, and by enhancing hepatic cholesterol degradation through CYP7a1 and CYP8b1 in mice fed a hypercholesterolemic diet.

Two Siblings with Cerebrotendinous Xanthomatosis / 대한피부과학회지

Cerebrotendinous xanthomatosis (CTX) is a rare autosomal recessive lipid storage disease caused by sterol 27-hydroxylase (CYP27) deficiency. We report two CTX siblings that were presented with typical manifestations such as achilles tendon xanthomas, mental retardation, progressive gait ataxia, and upper motor signs. Their parents and other three sisters were healthy. Serum cholesterol level was within normal limits for both siblings. The older brother has been treated conservatively with muscle relaxant and dopamine agonist because the disease was so progressive, but the younger sister has been treated with 250 mg/day chenodeoxycholic acid (CDCA) and 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor (simvastatin 40 mg/day) to prevent the progressive neurologic dysfunction.

The effect of fucoxanthin rich power on the lipid metabolism in rats with a high fat diet

This study determined the effects of fucoxanthin on gene expressions related to lipid metabolism in rats with a high-fat diet. Rats were fed with normal fat diet (NF, 7% fat) group, high fat diet group (HF, 20% fat), and high fat with 0.2% fucoxanthin diet group (HF+Fxn) for 4 weeks. Body weight changes and lipid profiles in plasma, liver, and feces were determined. The mRNA expressions of transcriptional factors such as sterol regulatory element binding protein (SREBP)-1c, Carnitine palmitoyltransferase-1 (CPT1), Cholesterol 7alpha-hydroxylase1 (CYP7A1) as well as mRNA expression of several lipogenic enzymes were determined. Fucoxanthin supplements significantly increased plasma high density lipoprotein (HDL) concentration (P < 0.05). The hepatic total lipids, total cholesterols, and triglycerides were significantly decreased while the fecal excretions of total lipids, cholesterol, and triglycerides were significantly increased in HF+Fxn group (P < 0.05). The mRNA expression of hepatic Acetyl-CoA carboxylase (ACC), Fatty acid synthase (FAS), and Glucose-6-phosphate dehydrogenase (G6PDH) as well as SREBP-1C were significantly lower in HF+Fxn group compared to the HF group (P < 0.05). The hepatic mRNA expression of Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) and Acyl-CoA cholesterol acyltransferase (ACAT) were significantly low while lecithin-cholesterol acyltransferase (LCAT) was significantly high in the HF+Fxn group (P < 0.05). There was significant increase in mRNA expression of CPT1 and CYP7A1 in the HF+Fxn group, compared to the HF group (P < 0.05). In conclusion, consumption of fucoxanthin is thought to be effective in improving lipid and cholesterol metabolism in rats with a high fat diet.

Las estatinas: Actividad biologica y produccion biotecnologica / Statins: Biological activity and biotechnological production

Las estatinas de tipo I son metabolitos f¨²ngicos de gran inter¨¦s no s¨®lo por su efecto hipocolesterolemiante sino por el n¨²mero de efectos pleiotr¨®picos que presentan. El papel de las estatinas en la reducci¨®n de l¨ªpidos en la sangre est¨¢ ampliamente documentado. En la actualidad, los estudios cl¨ªnicos han puesto en evidencia que las estatinas impactan positivamente en varios ¨®rganos y en diferentes estados de algunas enfermedades, independientemente de la reducci¨®n en los niveles de colesterol. Exhiben efecto antiinflamatorio, antioxidante y acci¨®n inmunomoduladora, lo que les confiere potencial impacto terap¨¦utico en el tratamiento de varias enfermedades. Estudios recientes las posicionan como antirretrovirales, impidiendo la replicaci¨®n del virus de inmunodeficiencia adquirida (VIH). El hecho de que generan pro-apoptosis, inhibici¨®n en el crecimiento y respuesta a favor de la diferenciaci¨®n de las c¨¦lulas neopl¨¢sicas de diversos or¨ªgenes, las hace ¨²tiles en el tratamiento de leucemia, c¨¢ncer de mama, c¨¢ncer colo-rectal, de pulm¨®n, de pr¨®stata y de p¨¢ncreas. No se deben dejar de lado los efectos ben¨¦ficos sobre el metabolismo ¨®seo, gracias a su asociaci¨®n con el aumento en la densidad mineral ¨®sea y el efecto protector contra el Alzheimer y otros tipos de demencia, posiblemente debido a su participaci¨®n en la relaci¨®n entre el ¦Â-amiloide y los niveles de colesterol.

Statins of type I are fungic metabollites of great importance not only by its hypocholestorolemic but also by its number pleoitropic effects that they show. The role of statins in the reduction of lipids in the blood is broadly reported. Currently, clinic surveys have put into evidence that statins both impact in a positive manner on several organs and in different stages of some diseases, regardless of the cholesterol reduction levels. They show an anti-inflammatory effect, anti-oxidant, and immune modulator action, which give them a therapeutic potential in the treatment of several diseases. Recent studies place them as anti-retrovirus avoiding the replication of the human immunodeficiency virus (HIV). The fact that they generate pro-apoptosis, inhibition on growing, and an favorable effect of the neo-plastic cell differentiation of diverse origins make them useful in the leukemia treatment, breast-cancer, colorectal, lung cancer, prostate cancer, and pancreas cancer. The beneficial side effects on bone metabolism should not be left aside due to its association with the increase of bone mineral density, the protector effect against Alzheimer and other types of dementia, possibly due to its participation in the relationship between ¦¢-amyloid and the cholesterol levels.

Gene Expression Analysis for Statin-induced Cytotoxicity from Rat Primary Hepatocytes

Statins are competitive inhibitors of hydroxy-3-methyl glutaryl coenzyme A (HMG-CoA) reductase and used most frequently to reduce plasma cholesterol levels and to decrease cardiovascular events. However, statins also have been reported to have undesirable side effects such as myotoxicity and hepatotoxicity associated with their intrinsic efficacy mechanisms. Clinical studies recurrently reported that statin therapy elevated the level of liver enzymes such as ALT and AST in patients suggesting possible liver toxicity due to statins. This observation has been drawn great attention since statins are the most prescribed drugs and statin-therapy was extended to a larger number of high-risk patients. Here we employed rat primary hepatocytes and microarray technique to understand underlying mechanism responsible for statin-induced liver toxicity on cell level. We isolated genes whose expressions were commonly modulated by statin treatments and examined their biological functions. It is of interest that those genes have function related to response to stress in particular immunity and defense in cells. Our study provided the basic information on cellular mechanism of statin-induced cytotoxicity and may serve for finding indicator genes of statin-induced toxicity in rat primary hepatocytes.

The Effects of Simvastatin on Bone Healing in Mandible Fractured Rats

PURPOSE: The hydroxymethylglutaryl coenzyme A reductase inhibitors (statins) are widely used in the treatment of dyslipidemia for lowering of cholesterol level. And the studies in simvastatins have shown to enhance bone formation in vitro and in vivo in rodents. But some other researchers have reported that simvastatins' anabolic effect on bone does not exist. The peripheral distribution beyond the liver represents a small fraction of an orally administered dose. We hypothesized that this poor peripheral distribution is the likely reason that simvastatins, yield ambiguous results as anabolic agents. We therefore investigated whether the effects of simvastatins on bone may be enhanced by subcutaneous administration, providing better peripheral delivery of these drugs. METHODS: 36 rat unilaterally mandible fractured models were prepared and divided into two groups. The simvastatin treated group where 1mg/kg of simvastatin was daily injected subcutaneously. The same dose of normal saline was injected on the control group. And 3 rats in each group were sacrificed and taken bone samples in each week. Bone sample was evaluated with tensile strength and histological morphology after 1, 2, 3, 4, 5 and 6 weeks. RESULTS: In simvastatin treated group, the fracture healing process, chondrocyte aggregation, collagen formation and trabecular bone formation was rapidly proceeded than the control group histologically. The tensile strength in the simvastatin treated group was measured as 1.02, 2.25, 3.95, 4.42, 5.49 and 6.00 N/mm2 each week, while it was 0.60, 1.05, 2.17, 3.75, 4.15 and 5.17 N/mm2 in the control group. The average tensile strength was higher by 1.04N/mm2 in simvastatin treated group. CONCLUSION: The currently available data on the effects of simvastatin on bone confirms that simvastatin helps fracture healing. And the potential for simvastatin to be used as anabolic agents for bone when delivered by the subcutaneous route.

Prospective Study to Evaluate the Efficacy and Safety of Pitavastatin in Patients with Risk Factor of Cardiovascular Disease(PEACE Study)

BACKGROUND AND OBJECTIVES: Pitavastatin, a recently approved synthetic 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, is known to effectively treat hypercholesterolemia. The goal of this study was to investigate the efficacy and safety of pitavastatin in hyperlipidemic Korean patients with coronary risk factors. SUBJECTS AND METHODS: This was an 8-week, prospective, multicenter, open-label clinical trial. The study subjects were hyperlipidemic Korean patients (triglyceride 130 mg/dL, age; 45-75 years) with at least two coronary risk factors. After a 2-week wash out period, the eligible subjects were given 2 mg of pitavastatin once daily for 8 weeks. In the case of the patients with LDL-cholesterol > or = 100 mg/dL after the first 4 weeks of treatment, the dose of pitavastatin was increased to 4 mg per day for the remaining 4 weeks. RESULTS: Of the 131 patients initially enrolled, 105 completed the study. Among the lipid profiles, the total cholesterol, triglyceride, and LDL-cholesterol levels showed a significant reduction with mean reduction rates of -30.66%, -23.92%, and -41.06%, respectively, after 8 weeks. Interestingly, the HDL-cholesterol level was significantly increased in the subjects with a low HDL-cholesterol level (HDL-cholesterol < 40 mg/dL) after 8 weeks of therapy (35.28+/-4.38 mg/dL to 40.39+/-6.45 mg/dL, 15.9%, p=0.001). The proportions of patients who achieved the LDL-cholesterol goal of the National Cholesterol Education Program Adult Treatment Panel III were 72.5% (37/51), 93.6% (44/47), and 100.0% (7/7) for the patients with goals of 100 mg/dL, 130 mg/dL, and 160 mg/dL, respectively. Five patients had mild adverse drug events, such as fatigue, itching, myalgia, and anorexia. No significant abnormalities were detected in the laboratory tests, including the liver function test and creatinine kinase level. CONCLUSION: The HMG-CoA reductase inhibitor, pitavastatin, was highly effective and generally well tolerated with an acceptable safety profile in hyperlipidemic Korean patients with coronary risk factors.