• Journal of Applied Biological Chemistry
• /
• 제59권1호
• /
• pp.49-56
• /
• 2016

The current study investigated the anti-obesity effect of Cissus quadrangularsis extracts (CQR-300) and its molecular action mechanism on obese mice induced high-fat diet (HFD). To induce the obesity, mice were fed a HFD for 6 weeks and then fed HFD only or HFD with CQR-300 at 50 and 200 mg/kg. Then, body weight gain and white adipose tissue weights were measured. We investigated the reduction in body fat and the regulation of fatty acid synthesis was measured by dual energy X-ray absorptiometry and real-time PCR with Western blot, respectively. In vitro study, CQR-300 inhibited pancreatic lipase activity. The CQR-300 treatment was significantly decreased the body weight gain and adipocytes size as well as white adipose tissues weights in HFD-induced obese mice. Furthermore, CQR-300 reduced the body fat and fat mass with regulating of adipose tissue hormones as leptin. Treatment with 50 mg/kg CQR-300 showed effectively lower expression levels of adipogenesis/lipogenesis related genes and proteins such as CCAAT/enhancer binding protein ${\alpha}$ ($C/EBP{\alpha}$), peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$), Sterol regulatory element binding protein-1c (SREBP-1c), and fatty acid synthase (FAS) in white adipose tissue (WAT) as compared with the HFD fed only mice. These results suggest that the CQR-300 has an anti-obesity effect via inhibition of lipase activity, decrease the body fat mass by regulating the adipogenesis and lipogenesis related genes and proteins in epididymal adipose tissue with evaluate body fat reduce in the HFD-induced obese mice.

• 약학회지
• /
• 제41권3호
• /
• pp.312-320
• /
• 1997

Pharmacokinetics and tissue distribution of DWP20349 and 20351, new quinolones, were examined in rats after a single intravenous and oral administration. Analyses of DWP20349 an d DWP20351 in plasma, tissue, and urine were determined by both HPLC and bioassay(microbiological assay). The plasma concentrations of the drugs declined biexponentially. The terminal half-lives ($t_{1/2\beta}$) of drugs were about 114 min (DWP20349) and 105 min (DWP20351) after intravenous dosing, and were 77 min (DWP20349) and 79 min (DWP20351) after oral dosing. The volume of distrbution at steady-state ($Vd_{ss}$) and total body clearances ($Cl_t$) of DWP20349 and DWP20351 were 760 ml/kg and 1126 ml/kg, and 5ml/min/kg and 10 ml/min/kg, respectively. The extents of bioavailability if DWP20349 and DWP20351 after oral administration were 29% and 28%, respectively. 24 h urinary recoveries measured by bioassay were 1.8% (DWP20349) and 1.3% (DWP20351) after oral dosing, and 2.4% (DWP20349) and 1.9% (DWP20351) after intravenous dosing. Plasma protein binding ratios ranged from 87%-90% (DWP20349) and 61%-68% (DWP20351). These drugs were highly distrbuted by the order of lung, kidney, liver and plasma (DWP20394), and lung, liver, kidney and plasma (DWP20351) after 1 hour orally administered.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권8호
• /
• pp.3767-3772
• /
• 2014

Introduction: MicroRNAs have emerged as post-transcriptional regulators that are critically involved in tumorigenesis. This study was designed to explore the effect of miRNA 133b on the proliferation and expression of TBPL1 in colon cancer cells. Methods: Human colon cancer SW-620 cells and human colon adenocarcinoma HT-29 cells were cultured. MiRNA 133b mimcs, miRNA 133b inhibitors, siRNA for TBPL1 and scrambled control were synthesized and transfected into cells. MiR-133b levels in cells and CRC tumor tissue was measured by real-time PCR. TBPL1 mRNA was detected by RT-PCR. Cell proliferation was studied with MTT assay. Western blotting was applied to detect TBPL1 protein levels. Luciferase assays were conducted using a pGL3-promoter vector cloned with full length of 3'UTR of human TBPL1 or 3'UTR with mutant sequence of miR-133b target site in order to confirm if the putative binding site is responsible for the negative regulation of TBPL1 by miR-133b. Results: Real time PCR results showed that miRNA 133b was lower in CRC tissue than that in adjacent tissue. After miR-133b transfection, its level was elevated till 48h, accompanied by lower proliferation in both SW-620 and HT-29 cells. According to that listed in http://www.targetscan.org, the 3'-UTR of TBPL1 mRNA (NM_004865) contains one putative binding site of miR-133b. This site was confirmed to be responsible for the negative regulation by miR-133b with luciferase assay. Further, Western blotting and immunohistochemistry both indicated a higher TBPL1 protein expression level in CRC tissue. Finally, a siRNA for TBPL1 transfection obviously slowed down the cell proliferation in both SW-620 and HT-29 cells. Conclusion: MiR-133b might act as a tumor suppressor and negatively regulate TBPL1 in CRC.

• BMB Reports
• /
• 제40권5호
• /
• pp.783-790
• /
• 2007

Receptor binding plays an important role in determining host specificity of the Bacillus thuringiensis Cry $\delta$-endotoxins. Mutations in domains II and III have suggested the participation of certain residues in receptor recognition and insect specificity. In the present study, we expressed the cloned domain II-III fragment of Cry4Ba and examined its binding characteristics to mosquito-larval midgut proteins. The 43-kDa Cry4Ba-domain II-III protein over-expressed in Escherichia coli as inclusion bodies was only soluble when carbonate buffer, pH 10.0 was supplemented with 4M urea. After renaturation via stepwise dialysis and subsequent purification, the refolded domain II-III protein, which specifically reacts with anti Cry4Ba-domain III monoclonal antibody, predominantly exists as a $\beta$-sheet structure determined by circular dichroism spectroscopy. In vitro binding analysis to both histological midgut tissue sections and brush border membrane proteins prepared from susceptible Aedes aegypti mosquito-larvae revealed that the isolated Cry4Ba-domain II-III protein showed binding functionality comparable to the 65-kDa full-length active toxin. Altogether, the data present the 43-kDa Cry4Ba fragment comprising domains II and III that was produced in isolation was able to retain its receptor-binding characteristics to the target larval midgut proteins.

• 한국식품영양과학회지
• /
• 제22권4호
• /
• pp.489-493
• /
• 1993

렉틴의 항영양작용기구 해명의 하나로서 마우스 반전소장에의 흡착양상을, $^3$H로 레벨한 렉틴을 가지고, 방사활성을 측정했다. 그 결과, 소장부위에 의한 흡착량은 유의차가 없었고, 흡착포화시간은 60분간 37$^{\circ}C$에서, 흡착량은 8.6$\times$$10^{7}$ 1mg intestine이 되고, 렉틴이 소장세포와 반응해서, 여러 가지 항영양작용을 유도한다고 생각된다. $^3$H 레벨한 렉틴 농도가 높아짐에 따라 흡착량은 크게되고, 2mg/$m\ell$에서 거의 포화에 달했다. pH7부근에서 흡착량이 제일 크고, 생체내에서의 소장은 렉틴이 가장 흡착되기 쉬운 상태임을 알 수 있었으며 저해인자는 fetuin과 EDTA이었다.

• Archives of Pharmacal Research
• /
• 제18권3호
• /
• pp.169-172
• /
• 1995

The changes in blood pressure may relate to the alterations of the responsiveness to vasoconstrictors and vasodilators, and these alterations can arise the modifications in the properties of angiotensin II (AII) receptor. In order to examine the changes of AII receptor in the hypertensive mechanism of renin-dependent hypertensive rats (RHRs; two-kidney, one-ligated type), we compared the equilibrium binding characteristics of $[^3H]$All in adrenal cortex and medulla from RHRs and normotensive rats. The dissociation constants of AII binding in both tissues of RHRs were very similar to those in the respective tissue of normotensive rats. However, the maximum binding was increased from 805 to 1050 fmole/mg protein in the adrenal cortex of RHRs, and decreased from 172 to 126 fmole/mg protein in the adrenal medulla of RHRs. These results imply that the up- and down-regulation of the All receptor population on the cell surface of adrenal glands from RHRs are consorted with the elevation of blood pressure and the activation of renin-angiotensin system.

• Journal of Animal Science and Technology
• /
• 제46권6호
• /
• pp.909-916
• /
• 2004

CCAAT/enhancer binding protein(C/EBP)는 지방전구세포의 분화과정에서 말현하는 전사인자군에 속한다. 이러한 C/EBP 중에서 C/EBP $\alpha$ 는 지방세포의 분화와 지방침착에 있어 중요한 역할을 수행허는 것으로 알려져 있다. 본 연구에서는 한우 C/EBP$\alpha$ 유전자를 분리 동정하고 그 발현을 조사하였다. 한우 C/EBP$\alpha$ 유전자는 1059bp의 open reading frame으로 구성되어 있으며 353개의 아미노산을 암호화하고 있었다. 그리고 한우 C/EBP $\alpha$ 아미노산을 다른 종과 비교하였을 때 매우 높은 상동성을 나타내었다. Northern blotting 분석에 의하여 12개월 한우에 있어서 C/EBP$\alpha$ mRNA의 분석을 실시한 결과 지방조직에서 가장 높은 말현을 나타내었으며 대장과 폐에서 매우 낮은 말현을 확인할수 있었다. 또한 12개월26개월 30개월 한우 의 등심과 지방에 있어서 C/EBP$\alpha$ 유전자의 발현을 real-time RT-PCR로 분석한 결과 등심 에서는 26개월에 서 가장 높은 발현을 보였으며 지방에서는 12개월과 26개월에 서 높은 발현을 나타내었다.

• 한방안이비인후피부과학회지
• /
• 제21권3호
• /
• pp.36-51
• /
• 2008

Objective : This study investigated the effects of PR(Pinelliae Rhizoma) on gene expression of lung tissue resected from asthma induced mice using intra-nasal instillation. Methods : Gene expression levels were measured using a microarray technique, and a functional analysis on these genes was conducted. Results : A total of 3270 genes were up-regulated or down-regulated, 860 genes which were lowered by induction of asthma were restored to those of naive animals, Furthermore hand, 1235 genes were lowered to normal levels, which were elevated by induction of asthma. Most of changed genes were involved in signalling pathways. Genes in which expression levels were restored by oral administration of PR were involved in MAPK pathway, focal adhesion, and regulation of actin cytoskeleton etc. Genes of which expression levels were lowered by oral administration of PR were involved in rhodopsin-like receptor activity, zinc ion binding and ATP binding. These genes were also involved in neuroactive ligand receptor interaction, the JAK-STAT signaling pathway and also the T-cell receptor signaling pathway. Conclusion : These results demonstrate the strong possibility that the mechanisms of PR on asthma are involved in neuroactive ligand receptor interaction pathway or related molecules.

• 약학회지
• /
• 제39권3호
• /
• pp.223-230
• /
• 1995

The phannacokinetics and tissue distribution of DWQ-013, a new quinolone, were examined in rats and mice following a single intravenous and oral administration. DWQ-013 in plasma and urine was determined by both HPLC and microbiological assay. The plasma concentration of the drug declined biexponentially. The terminal half life of the drug was 11.11$\pm$0.14 hour after intravenous dosing. The volume of distribution at terminal elimination phase(Vd$_\beta$) and total clearance of the drug were 1.29$\pm$0.15 l/kg and 0.78$\pm$0.09 l/h/kg. The bioavailability of DWQ-013 after oral administration was 56.0% (HPLC) and 77.2%(bioassay), respectively. Twelve-hour urinary recovery of drug was measured by HPLC and bioassay to 0.035$\pm$0009% and 4.71$\pm$066% after oral dosing, to 0.055$\pm$0.014% and 7.65$\pm$1.53% after intravenous dosing, which may indicate the presence of biologically active metabolites. Binding of the drug to plasma proteins ranged from 97%~99% at various concentrations. The drug was highly distributed in order of liver, kidney and lung after 1.5 hours in mice.

• Journal of Plant Biology
• /
• 제38권4호
• /
• pp.343-348
• /
• 1995

Treatment of Pisum sativum tissue with the protein kinase inhibitor staurosphorine resulted in impairment of 3H-indoleacetic acid transport in etiolated stem segments. The transport inhibitiion was accompanied by an increase in net uptake of labeled auxin in the tissue. The magnitude of auxin accumulation in tissue treated with the phytotropin N-1-naphthylphthalaic acid (NPA) which specifically blocks the efflux of auxin in the plasma membrane was reduced by the protein kinase inhibitor, suggesting that inhibition of protein phosphorylation could lead to hindrance of the auxin-exporting function of NPA receptors. The flavonoid genistein which is also known to inhibit protein kinase likewise reduced NPA-induced auxin accumulation. However, the flavonoid did not bring about auxin accumulation by itself, nor did it inhibit auxin transport. In view of the finding that the flavonoid also competes with NPA for a common binding site, a mechanism for the flavonoid effect on the NPA action will be proposed.