T 세포 및 대식세포 기능에 대한 Silybin의 조절효과

조재열,Cho, Jae-Youl 대한약학회 2007 약학회지 Vol.51 No.4

Silybin is known to be a major active flavonoid component isolated from Silybum marianum, a hepatoprotective medicinal plant. In this study, we examined the immunomodulatory role of silybin on T cell and macrophage-mediated immune responses. To do this, the proliferation of splenic lymphocytes and CD8+ CTLL-2 cells under mitogenic stimulation with lipopolysaccharide (LPS), concanavalin (Con) A and interleukin (IL)-2 and the production of $TNF-{\alpha}$ and NO from LPS- and $IFN-{\gamma}$-activated macrophages was evaluated under silybin treatment. The mitogenic proliferation of splenic lymphocytes induced by LPS and Con A was strongly diminished by silybin in a dose-dependent manner. Moreover, the proliferation of CD8+ CTLL-2 cells was also negatively modulated by the compound. In contrast, silybin did not strongly suppress the proliferation of normal splenocytes and T cell line Sup-T1 cells, indicating that the inhibitory effect of silybin may be due to blocking only mitogenic responses of splenic lymphocytes. In addition, silybin inhibited $TNF-{\alpha}$ production in LPS-stimulated RAW264.7 cells. Effect of silybin however was distinct, according to NO-inducing stimuli. Thus, silybin only blocked NO production induced by $IFN-{\gamma}$ but not LPS and the inhibition was increased when PMA was co-treated with $IFN-{\gamma}$. Unlike NO inhibition, however, this compound protected the cytotoxic damage of RAW264.7 cells induced by both LPS and $IFN-{\gamma}$. Therefore, our data suggest that silybin may participate in host immune responses mediated by T cells and macrophages via regulating mitogenic proliferation, and the production of $TNF-{\alpha}$ and NO, depending on cellular stimuli.

S. abortus 유래 LPS와 E. coli 유래 LPS에 의한 패혈증성 쇽 유도 작용 비교

조재열,유은숙,Cho, Jae-Youl,Yoo, Eun-Sook 대한약학회 2007 약학회지 Vol.51 No.1

Acute septic shock is one of inflammatory diseases mediated by pro-inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$. In this study, we examined the pathological difference and mechanism of lipopolysaccharides isolated from E. coli (E-LPS) or S. abortus (S-LPS) on inducing acute septic shock in ICR mouse. All mice were died by intraperitoneal treatment of S-LPS with 0.75 mg/kg, whereas E-LPS treated with even 3 mg/kg only showed 30% of mice lethal, indicating that S-LPS may be more feasible in triggering a strong septic shock condition. The secretion pattern of TNF-${\alpha}$, a critical pro-inflammatory cytokine in septic shock condition, was also distinct between E-LPS- and S-LPS-treated groups. Thus, S-LPS strikingly increased serum level of TNF-${\alpha}$ (6 ng/ml) at 1 h, while E-LPS just displayed at 2 ng/ml level. However the interaction of S-LPS with LPS receptor toll like receptor (TLR)-4, was not stronger than that of E-LPS, according to experiments with macrophage cell line RAW264.7 cells. Thus, E-LPS rather than S-LPS strongly enhanced the production of TNF-${\alpha}$. Interestingly, S-LPS more strongly up-regulated splenocyte proliferation, compared to E-LPS group, whereas there was no difference between S- or E-LPS treated groups in proliferation of Balb/c- or C57BL/6-originated splenic lymphocytes. Therefore, our data suggest that S-LPS is a more active endotoxin and that the strong septic shock-inducing effect of S-LPS seems due to the enhancement of early TNF-${\alpha}$ production and S-LPS-sensitive lymphocyte proliferation.

간 보호제 및 담즙산류들이 마크로파지 세포주에서 TNF-alpha 분비에 미치는 효과

조재열(Jae Youl Cho),박지수(Ji Soo Park),유은숙(Eun Sook Yoo),백경업(Kyong Up Baik),박명환(Myung Hwan Park) 대한약학회 1998 약학회지 Vol.42 No.1

The effect of hepatoprotective agents and bile acids on tumor necrosis factor-alpha, (TNF-alpha) production in murine and human macrophage cell line (RAW264.7 and U937) was investigated. The hepatoprotective agents including silymarin and its major component, silybin, significantly inhibited TNF-alpha production in a concentration dependent manner (IC50 of silybin=67.7mcg/ml (140.3mcM)). In differentiated U937 cells, especially, silybin showed more effective inbitory activity (IC50=35.1mcg/ml (72.7mcM)). These results suggest that silymarin and silybin may inhibit TNF-alpha production in the process of hepatic diseases in human. However, biphenyldimethyl dicarboxylate (DDB) was not effective. In the case of bile acids, chenodeoxycholic acid (CDCA) showed a concentration dependent inhibitory effect on TNF-alpha production (IC50 of CDCA= 71.5mcg/ml (182.1mcM)). In contrast, glycine or taurine conjugated form (G-CDCA or T-CDCA) restored to the control level or significantly increased TNF-alpha production. And also ursodeoxycholic acid (UDCA) and its conjugated forms (G-UDCA and T-UDCA) showed a variety of patterns on TNF-alpha production by changes of functional groups and concentration. These results also indicate that bile acids may regulate TNF-alpha production in normal hepatic function or disease conditions.

인삼 사포닌류가 종양괴사인자의 생성 및 T 세포 증식에 미치는 효과

조재열(Jae Youl Cho),박지수(Ji Soo Park),유은숙(Eun Sook Yoo),백경업(Kyong Up Baik),박명환(Myung Hwan Park),한병훈(Byung Hoon Han) 대한약학회 1998 약학회지 Vol.42 No.3

To investigate the effects of ginsenosides from Panax ginseng on mitogenic responses in macrophages and splenocytes from murine, we examined the effects of representative protopanaxadiol and protopanaxatriol ginsenosides (Rb1, Rb2, Re and Rg1) on tumor necrosis factor-alpha (TNF-(alpha) production in and smurine macrophage cell line (RAW264.7 cells) stimulated by lipopolysaccharide (LPS) and T cell proliferation in splenocytes stimulated by concanavalin A (Con A). Among the ginsenosides tested, protopanaxadiol ginsenosides (Rb1 and Rb2) significantly inhibited TNF-alpha production in a dose-dependent manner. However, protoppanaxatriol ginsenosides (Re and Rg1) showed little inhibitory activity. The molar concentrations of Rb1 and Rb2 producing 50% inhibition (IC50) of TNF-alpha production were 55.8mcg/ml (48.0mcM) and 31.8mcg/ml (27.9mcM), respectively. As a positive control, prednisolone also exhibited inhibitory activity with an IC50 value of 21.7mcM. In T cell proliferation, Rg1, was not effective but Rb1 and Re or Rb2 significantly increased or inhibited at high concentration, 75 and 100mcg/ml. In contrast, prednisolone showed potent inhibitory activity with an IC50 value of 6.1nM. These results suggest that ginsenosides may take part in the mitogen-induced signaling pathway for TNF-alpha production and T cell proliferation from macrophages splenocytes.

Ginsenoside Rg3 및 그 유도체 Ginsenoside Rg3-2H의 NO 생성 및 lymphocyte 분열 억제 효과

조재열(Jae Youl Cho) 고려인삼학회 2008 Journal of Ginseng Research Vol.32 No.3

Ginsenosides are major components in Panax ginseng and known to have numerous pharmacological activities such as anti-cancer, anti-diabetes, anti-viral and anti-atherosclerosis effects. In this study, the regulatory activities of G-Rg3 and its derivative 25-hydroxy Rg3 (G-Rg3-2H) on the production of nitric oxide (NO) in macrophages and the proliferation of lymphocytes prepared from spleen and bone marrow under treatment of lipopolysaccharide (LPS) or concanavalin (Con) A were examined. G-Rg3 and G-Rg3-2H dose-dependently inhibited NO production from LPS-activated RAW264.7 cells and in agreement, these compounds protected RAW264.7 cells from LPS-mediated cytotoxicity. In contrast, G-Rg3-2H dose-dependently inhibited lymphocyte proliferation induced by both LPS and Con A, while there was no inhibition by G-Rg3. Therefore, our data suggest that these compounds may be applied for NO-mediated or lymphocyte- mediated immunological diseases.

돼지 간 및 정소에서 단백질 카르복실메칠화 현상

조재열(Jae Youl Cho),김성수(Hyang Woo Lee),이향우(Hyang Woo Lee),홍성렬(Sungyoul Hong) 대한약학회 2001 약학회지 Vol.45 No.1

Protein carboxy O-methylation is a kind of enzymatic reaction producing carboxy methylester catalyzed protein carboxyl O-methyltransferases at the carboxyl group of amino acid residues in polypeptide. Since the finding of carboxyl methylester, many studies have been focused on the understanding of biological functions in eukaryotes but still not clear except for roles in Ras attachment to membrane and protein repair. In this study, we investigated the protein carboxyl methylation in porcine liver and testis in repect of identification and characterization of carboxyl methylesters and natural proteinous substrates using pH stability of the esters and electrophoresis under acidic and bacis conditions. We detected several kinds of methyl esters, 3 kimds each in cytosolic fractions from liver and testis. Under the treatment of strong acid and base, the ratio between base- stable substrates and unstable ones in liver (4:6) was different from the ratio obtainde in testis (6:4). The methyl accepting capacities were affected by enzymatic proteolysis between the range of 55 to 65% in liver and of 35 to 45% in testis. Separation of the methylated proteins by acidic elelctrophoresis in the presesnce of urea and SDS revealed distinctively natural substrates of 26, 33 and 80kD in the cytosol from liver, and of 14, 25, 32 and 86kD from testis. Most of the labelling, however, were lost following electrophoreses under moderare alkaline condition, except fot molecules of newly detected 7 and 17kD in liver, and 15, 29, 40 and 80kD in testis. From these results, it was proposed that protein carboxyl O-methylation in each organs may be catalyzed by different classes of protein carboxyl O- methyltransferases. In addition, it is suggested that the protein carboxyl methylation in liver and testis may have different patterns in respect of natural substrates.

C7- 이환체 구조를 갖는 새로운 플루오로퀴놀론계 항생물질의 흰쥐 체내동태와 조직분포

조재열(Jae Youl Cho),한승희(Seung Hee Han),김병오(Byoung O Kim),남권호(Kweon Ho Nam),손호정(Ho Jung Son),이재욱(Jae Wook Lee),유영효(Young Hyo Yu),박명환(Myung Hwan Park) 한국응용약물학회 1997 Biomolecules & Therapeutics(구 응용약물학회지) Vol.5 No.3

The pharmacokinetics of DWP20364 (1-cyclopropyl-5-amino-6,8-difluoro-7-(2,7-diazabicyclo[3,3,0] oct-4-ene-7-yl)-1,4-dihydro-4-oxoquinoline-3-carboxylic acid), a novel fluoroquinolone containing C7-bicyclic structure, were compared with those of ciprofloxacin (CPFX) after single intravenous (i.v.) and oral (p.o.) administration to rats using microbiological assay (bioassay). After i.v. administration to rats, the plasma concentrations of the two drugs declined biexponentially. The terminal half-lives (t_(½β)) of DWP20364 were 110±13.2 min and 117±3.09 min after i.v. and p.o. administration, respectively, and they were significantly higher than those of CPFX (45.5±9.52 min and 48.3±12.1 min, respectively). Similar results were also obtained from plasma concentrations and area under the plasma concentration-time curves. The total body clearance of DWP20364, 7.82±0.37 ml/min/kg was significantly slower than that of CPFX, 27.3±11.1 ml/ min/kg. Above data suggested that the antimicrobial activity of DWP20364 could be lnger than that of CPFX. The urinary recovery after i.v. and p.o. administration of DWP20364 was signifcantly lower than those of CPFX suggesting that the effect of DWP20364 on urinary tract infection could be lower than that of CPFX. The serum protein binding values of DWP20364 at 2 ㎍/ml were apparently 91.5∼93.1% in rats and human. DWP20364 was distributed by the order of liver, lung, kidney, spleen, heart, muscle and brain collected at 30 min after orally administered.

C7위치에 3-아미노-4-메칠치오메칠피로리디닐기를 치환한 신규 퀴놀론계 항생물질 DWP10349 및 DWP20351의 흰쥐에서의 체내동태 및 조직분포

조재열(Jae Youl Cho),남권호(Kweon Ho Nam),유은숙(Eun Sook Yoo),이재욱(Jae Wook Lee),유영효(Young Hyo Yu),박명환(Myung Hwan Park) 대한약학회 1997 약학회지 Vol.41 No.3

Pharmacokinetics and tissue distribution of DWP20349 and 20351, new quinolones, were examined in rats after a single intravenous and oral administration. Analyses of DWP20349 and 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 (t1/2beta) 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 (Vdss) and total body clearances (Clt) 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.

강활에서 종양괴사인자 생성을 억제하는 활성성분의 분리

조재열(Jae Youl Cho),이종수(Jong Soo Lee),박지수(Ji Soo Park),박명환(Myung Hwan Park) 大韓藥學會 1998 약학회지 Vol.42 No.2

The total ethanol extract of Angelica koreana radix (UmbeUiferae) showed potent inhibitory effect on tumor necrosis factors (TNF-alpha) production from LPS-stimulated RAW264.7 cells in screening studies on 118 Korean medicinal plants. The activity-guided fractionation of the total ethanol extract resulted in the isolation of 4 compounds, including active substances. The chemical structures of the compounds isolated were established by chemical and spectrometric analyses as imperatorin, isoimperatorin, osthol and oxypeucedanin. Among them, osthol, oxypeucedariin and isoimperatorin showed significant inhibitory activities on TNF-alpha production as 54.3+/-2.1%, 33.6+/-1.0% and 17.7+/-3.0%, respectively, at 12.5mcg/ml.

흰쥐에서 Ursodeoxycholic Acid 및 Silymarin을 함유한 의약조성물(DWP305)의 연용투여에 의한 간내 담즙산 조성변화

조재열(Jae Youl Cho),연제덕(Je Deuk Yeon),남권호(Kweon Ho Nam),김점용(Jeum Yong Kim),유은숙(Eun Sook Yoo),유영효(Young Hyo Yu),박명환(Myung Hwan Park) 대한약학회 1996 약학회지 Vol.40 No.3

DWP305, a preparation containing combination of ursodeoxycholic acid(UDCA), silymarin and vitamins (B1 and B2), is a drug currently being developed for hepatic disorders. In order to evaluate the changes in hepatic function by multiple oral administration(2 and 4 weeks) of DWP305 in rats, several biochemical parameters in blood, bile acid composition, and the accumulation of UDCA and lithocholic acid(LCA),a toxic metabolite formed by enterobacteria, were examined using HPLC. In blood biochemical findings, DWP305 did not affect the normal level and there was no difference in total bile acid composition for UDCA, cholic acid(CA), deoxycholic acid(DCA), chenodeoxycholic acid(CDCA) and LCA compared to the UDCA administered group, although total ratio of UDCA and CA was different from normal group. In case of ratio of taurine and glycine conjugated forms, DWP305(186mg/kg as a UDCA) administered group was also similar to normal group and UDCA administered group, while high dosing of DWP305 was not different in the ratio of UDCA administered group(930mg/kg) but normal group. And the ratio of LCA was in order of UDCA(930mg/kg), DWP305(930mg/kg as a UDCA), UDCA(186mg/kg) and DWP305(186mg/kg as a UDCA) administered group, which was less than 4%. The free form of UDCA as well as most of bile acids was not detected at all in rat liver, indicating that there''s no accumulation. These results suggest that multiple dosing of DWP305 in rats may not affect hepatic biotransformation and metabolism of bile acids.