Low-grade pro-inflammatory state and leptin resistance are important underlying mechanisms that contribute to obesity-associated hypertension. We tested the hypothesis that Astragaloside IV (As IV), known to counteract obesity and hypertension, could prevent obesity-associated hypertension by inhibiting pro-inflammatory reaction and leptin resistance. High-fat diet (HFD) induced obese rats were randomly assigned to three groups: the HFD control group (HF con group), As IV group, and the As IV + ${\alpha}$-bungaratoxin (${\alpha}-BGT$) group (As IV+${\alpha}-BGT$ group). As IV ($20mg{\cdot}Kg^{-1}{\cdot}d^{-1}$) was administrated to rats for 6 weeks via daily oral gavage. Body weight and blood pressure were continuously measured, and NE levels in the plasma and renal cortex was evaluated to reflect the sympathetic activity. The expressions of leptin receptor (LepRb) mRNA, phosphorylated signal transducer and activator of transcription-3 (p-STAT3), phosphorylated phosphatidylinositol 3-kinase (p-PI3K), suppressor of cytokine signaling 3 (SOCS3) mRNA, and protein-tyrosine phosphatase 1B (PTP1B) mRNA, pro-opiomelanocortin (POMC) mRNA and neuropeptide Y (NPY) mRNA were measured by Western blot or qRT-PCR to evaluate the hypothalamic leptin sensitivity. Additionally, we measured the protein or mRNA levels of ${\alpha}7nAChR$, inhibitor of nuclear factor ${\kappa}B$ kinase subunit ${\beta}/nuclear$ factor ${\kappa}B$ ($IKK{\beta}/NF-KB$) and pro-inflammatory cytokines ($IL-1{\beta}$ and $TNF-{\alpha}$) in hypothalamus and adipose tissue to reflect the anti-inflammatory effects of As IV through upregulating expression of ${\alpha}7nAChR$. We found that As IV prevented body weight gain and adipose accumulation, and also improved metabolic disorders in HFD rats. Furthermore, As IV decreased BP and HR, as well as NE levels in blood and renal tissue. In the hypothalamus, As IV alleviated leptin resistance as evidenced by the increased p-STAT3, LepRb mRNA and POMC mRNA, and decreased p-PI3K, SOCS3 mRNA, and PTP1B mRNA. The effects of As IV on leptin sensitivity were related in part to the up-regulated ${\alpha}7nAchR$ and suppressed $IKK{\beta}/NF-KB$ signaling and pro-inflammatory cytokines in the hypothalamus and adipose tissue, since co-administration of ${\alpha}7nAChR$ selective antagonist ${\alpha}-BGT$ could weaken the improved effect of As IV on central leptin resistance. Our study suggested that As IV could efficiently prevent obesityassociated hypertension through inhibiting inflammatory reaction and improving leptin resistance; furthermore, these effects of As IV was partly related to the increased ${\alpha}7nAchR$ expression.
The effect of ginsenoside-Rb2, one of a major pharmacological component of Panax ginseng C.A. Meyer, on low density lipoprotein (LDL) receptor expression was investigated and compared with hypocholesterolemic drug lovastatin. In HepG2 cell, exogenous cholesterol decreased LDL receptor mRNA expression, but ginsenoside-Rb2 recovered this reduction of LDL receptor mRNA up to normal expression level. Lovastatin also increased LDL receptor mRNA expression as similar as ginsenoside-Rb2 did. The reduction of sterol regulatory element binding protein (SREBP) transcription by exogenous cholesterol was also similarly recovered by ginsenoside-Rb2 and lovastatin addition. Compound K, a metabolite of ginsenoside-Rb2 and -Rb1 by human intestinal bacteria also increased the SREBP mRNA expression in cholesterol-enriched condition. Ginsenoside-Rb2 seems to up-regulate LDL receptor mRNA expression through the induction of de novo SREBP transcription. Therefore, increased expression of SREBP mRNA by ginsenoside-Rb2 elevated the LDL receptor mRNA expression in HepG2 cells, and these inductions possibly drop the plasma cholesterol level in hypercholesterolemia patients, in vivo, as likely in case of lovastatin.
Park, Jung-Sun;Kim, Hye-Kyung;Lee, Hye-Won;Lee, Mi-Hyun;Kim, Hyun-Gi;Chae, Soo-Wan;Chae, Han-Jung
Korean Journal of Clinical Pharmacy
/
v.16
no.1
/
pp.23-27
/
2006
Doxorubicin (DXR) is a type of anti-cancer drug called an 'anthracycline glycoside', It works by impairing DNA synthesis, a crucial feature of cell division, and thus is able to target rapidly dividing cells. Doxorubicin is a very serious anti-cancer medication with definite potential to do great harm as well as great good. A liquid chromatography-tandem mass spectroscopy (LC-MS/MS) method was developed to identify and quantify DXR in small-volume biological samples. After the addition of internal standard (IS, $5{\mu}L\;of\;1{\mu}M/ml$ daunorubicin methanol solution) into the serum sample, the drug and IS were extracted by methanol. Following vortex for a 1min and centrifugation at 15,000g for 10 min the organic phase was transferred and evaporated under a vacuum. The residue was reconstituted with $350{\mu}L$ of mobile phase and $10{\mu}L$ was injected into C18 column with mobile phase composed of 0.05M ammonium acetate (0.1 M acetic acid adjusted to pH 3.5) and acetonitrile (40:60, v/v). The flow rate was kept constant at $350{\mu}L/min$. The ions were quantified in the multiple reaction mode (MRM), using positive ions, on a triple quadrupole mass spectrometer. The lower limits of quantification for Doxorubicin in plasma and small tissues were approximately 0.5 ng/mL and 0.5 ng/mL respectively. Intra- and inter-assay accuracy (% of nominal concentration) and precision (% CV) for all analytes were within 15%, respectively.
Kim, Jae-Gon;Lee, Young-Soo;Baik, Byeong-Ju;Park, Byung-Keon
Journal of the korean academy of Pediatric Dentistry
/
v.28
no.3
/
pp.447-463
/
2001
Dendroaspis natriuretic peptide (DNP), a fourth member of the natriuretic peptide isolated from the venom of the Dendroaspis angusticeps snake, has been reported to be present in human plasma and atrial myocardium and caused vasorelaxation and diuresis in experimental animals. However, it is uncertain whether they are present in peripheral organs other than the heart and its further physiological roles also remains to be clarified. To assess the possible physiological role of DNP in the salivary glands, I investigated the localization of DNP peptide in the rat salivary glands by immunohistochemistry and the binding sites for radiolabelled DNP in the rat salivary glands and oral mucosa using in vitro autoradiography. DNP immunoreactivity was widely distributed in the submandibular, sublingual and parotid glands, particularly in the ducts such as the intercalated and striated ducts, where atrial natriuretic peptide (ANP) was colocalized in consecutive sections, but not in acini. High density $^{125}I-DNP$ binding sites were localized in the epithelia of the tongue and hard palate, while low density binding sites for $^{125}I-DNP$ were also distributed in the submandibular, sublingual, and parotid glands. In the hard palate and tongue, the precise location of this binding was revealed on the basal and parabasal cells of the epithelia by emulsion microautoradiography. These results suggest that DNP may not only have a role in the salivary glands but also play a role in the regulation of growth in the oral epithelium, particularly in the hard palate and tongue.
Journal of the Korea Academia-Industrial cooperation Society
/
v.21
no.4
/
pp.392-399
/
2020
This study was undertaken to determine the effects of feed supplementation with Hermetia illucens (HI) on the fecal score, blood profiles, immune response, and small intestinal morphology in weaned pigs. A total of 24 weaned pigs (Landrace×Yorkshire×Duroc; 5.86±0.13 kg) were randomly allotted to 4 treatments and 6 replicates on the basis of initial body weight. The experiment was accomplished over 0-14 days. The dietary treatments included a corn-soybean meal diet supplemented with 0, 1, 2, and 3% HI. A linear response to increasing dietary HI was observed for the number of monocytes (p<0.01) and eosinophils (p<0.05), whereas red blood cells tended to decrease with increasing HI levels. Plasma TNF-α levels were also determined to linearly decrease with HI supplementation (p=0.07). Moreover, a linearly decreasing tendency (p=0.06) was observed in the fecal score with increasing dietary levels of HI. Weaned pigs fed diets supplemented with increasing dietary concentrations of HI showed linearly improved (p<0.05) duodenal villus height during the study period. Taken together, these results indicate the beneficial effects of HI on diarrhea reduction, immune response, and small intestinal morphology in weaned pigs.
Lee, . H. C.;S. J. Uhm;S. Y. Ann;H. J. Chung;Park, H. D.;Lee, H. T.;K. S. Chung
Korean Journal of Animal Reproduction
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v.24
no.4
/
pp.439-449
/
2000
This study was to investigate the expression of transgene after co-injection of spermatozoon and EGFP gene into mature oocytes in cattle. From frozen semen, spermatozoa were treated by DTT with 0.03% Tween-20, freezing and thawing or 0.02% Triton X-100 to disrupt their plasma membranes. The sperm injected oocytes were co-cultured with bovine oviduct epithelial cells in CRlaa, and expression of EGFP in embryos were observbed under epifluorescent microscope. Two pronuclei were formed in oocytes injected with sperm treated by DTT (44.6%), DTT-Tween-20 (48.4%), DTT-freezing (44.4%) and DTT-Triton X-100 (42.9%). Cleavage and blastocyst formation rates of bovine oocytes which injected with sperm treated by DTT, DTT-Tween-20, DTT-freezing, and DTT-Triton X-100 were 49.1, 58.5, 43.9, and 48.4% and 10.2, 13.0, 6.8, and 6.5%, respectively. Although the most of embryos were showing mosaicism, embryos expressing EGFP gene were observed in all treated groups. Therefore, these results indicate that membrane disrupted sperm could interact with exogenous DNA, and that this procedure may be useful to introduce foreign gene into bovine oocytes.
Kim Jeong Jin;Yang Sung Wan;Son Nak Won;Ahn Kyoo Seok
Journal of Physiology & Pathology in Korean Medicine
/
v.17
no.2
/
pp.428-435
/
2003
This research was performed to examine an anti-inflammatory effects of Gamisangryosamul-Tang(GSS) and anti-pruritus effects of Ziyang-Go(Salve). This study was processed by three experiments; Experiment 1: Inhibitory activity of GSS extract on the degranulation of mast cell and histamine release in plasma induced by compound 48/80 i.p, injection after the pretreatment of GSS extract i.p, injection in Sprague-Dawley rats, Experiment 2: Anti-inflammatory effect of GSS extract on macropharge raw 264,7 cells treated by LPS 250 ppm (before 2 hours). Experiment 3: Measurement of passive cutaneous anaphylaxis and atopic dermatitis using NC/Nga mice, GSS extract inhibited histamine release by 70% compared to compound 48/80 treated control group and histologically significantly reduced (P<0,01) the degranulation of mast cell in SD rats. In GSS extract treated group, the expression of TNF-α in macropharge cell showed the remarkable inhibitory effect about 62% (P<0,01) compared to LPS treated control group. The expression of IL-6 appeared more effective by 46% than the LPS treated control group and by 6% compared to hydrocortison treated group, Comparing with steroid (0.05% prednisolon) ointment, Ziyan-Go treated group showed the significant(30%) recovery on skin response index in atopic dermatis like anaphylaxis mice(NC/Nga), Finally, in scratching behavioral tests of NC/Nga mice for three weeks, Ziyang-Go treated group significantly (P<0.05) suppressed the pruritus on the face, neck, ears and dorsal skin than inbred NC/Nga mice. However, the change of IgE and IFN-γ from the spleen cell of NC/Nga mice was not significantly different between the oral intake of GSS extract group and of saline intaked control group. Summary and Conclusion: This study demons trates that Ziyang-go have the equal anti-pruritus effect to steroid ointment and GSS extract have the notable immunologic activity on inflammatory in vivo and in vitro model. Advanced experiment of this study will be required for more reliable information about the correlation between the lymphokine (i.e. IgE) and the anti-allergic effects of GSS.
Inflammation scan using radiolabelled leukocytes has high sensitivity and specificity. Several methods for labelling leukocytes have been evaluated using P-32 diisopropyl fluorophosphate (DFP-32), H-3 thymidine, Cr-51 chromate, Ga-67 citrate and Tc-99m-sulfur colloid. In-111-oxine has proved so far to be the most reliable agent for labelling leukocytes. In-111-oxine is, however, expensive, not easily available when needed, and its radiation dose to leukocytes is relatively high. Moreover, resolution of the resultant image is relatively poor. Tc-99m is still the agent of choice because of, as compared with the indium, its favorable physical characteristics, lower cost and availability. Now the technique for labelling the leukocytes with technetium is successfully obtained using the lipophilic HAPAO with higher efficiency for granulocytes than for other cells. With this technique it is possible to label leukocytes in plasma to improve the viability of the leukocytes. Inflammation scan using Tc-99m-HMPAO has been evaluated in several laboratories, and difference in methods for separation and labelling accounts for difference in efficiency, viability and biodistribution of the labelled leukocytes. We performed inflammation scan using leukocytes labelled with Tc-99m-HMPAO in three dogs 24 hours after inoculation of live E. Coli and A. Aureus in their right abdominal wall. We separated mixed leukocytes by simple sedimentation using 6% hetastarch (HES) and labelled the leukocytes with Tc-99m-HMPAO in 20% cell free plama diluted with phosphate buffer solution(Fig. 1). Uptake was high in the liver and spleen but is was minimal in the lungs on whole body scan. Kidneys and intestine showed minimal activity although it was high in the urinary bladder(Fig. 2). Uptake of labelled leukocytes in the inflammation site was do(mite on 2 hour-postinjection scan and abscess was clearly delineated on 24 hour-delayed scan with high target-to-nontarget ratio(Fig. 3, 4). Inflammation scan using mixed leukocytes labelled with Tc-99m-HMPAO is very sensitive and specific in early detection of inflammation.
We investigated the effect of Perilla frutescens (PF) ethanol extract powder (PF-E30) on the local allergic reaction activated by anti-DNP IgE and the mast cell-mediated immediate-type allergic reactions induced by compound 48/80 in a mouse model. One gram of PF powder extracted with 30% ethanol at $80^{\circ}C$ contained 12.3 mg of rosmarinic acid. Oral administration of PF-E30 (0.1 to 0.5 mg/kg body weight) significantly reduced plasma histamine levels and inhibited histamine release from peritoneal mast cells in mice activated by compound 48/80 or anti-DNP IgE. Moreover PF-E30 dose-dependently inhibited the production of antigen-induced IgE. These results indicate that the PF ethanol extract inhibits mast cell-mediated immediate-type allergic reactions in vivo and in vitro.
The Journal of the Korean Society for Microbiology
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v.21
no.2
/
pp.211-226
/
1986
To understand the pathogenesis of anicteric leptospirosis with severe pulmonary hemorrhage occured in Korea, the microbiological and pathological features were observed in the experimentally induced leptospirosis in guinea pigs infected with a virulent strain of Leptospira interrogans isolated from the patient at Wonju, Korea, and the results are summarized as follows. 1. The main pathological features were widespread hemorrhages, especially affecting lung, skeletal muscles, retroperitoneal and perirenal adipose tissues. The hemorrhages accompanied inflammatory process especially of vasculitic pattern as well as occasional coagulation necrosis in the liver, skeletal muscle, and myocardium. The main inflammatory cells were of plasma cell even in the fairly early stage of the infection. 2. Those pathologic changes were more exaggerated in the inoculation site. 3. Within 144 hours of infection, the longer the infection time, the more antigens were observed in the tissues, and the severer the pathologic changes. 4. Leptospiral antigens were detected at first by indirect immunofluorescent and immunoperoxidase technics. As the infection time extended, the antigens were observed in all of the tissues examined except in the skeletal muscle. The shape of the antigens was spiral or thread-like within 72 hours of infection. As the infection progressed, they became fragmented and granular. 5. Leptospires were detected in the blood within 144 hours of infection by darkfield microscopic examination. Thereafter, none was observed. 6. Antibody to leptospires were detected as early as 72 hours of infection. In summary, the virulent strain of L. interrogans used in this experiment induced widespread hemorrhages with inflammatory reaction especially in lung, skeletal muscles, and retroperitoneal adipose tissue. With these findings, it is suggested that the direct toxic effect of leptospires might playa great role in the pathogenesis of this infection.
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