• 생명과학회지
• /
• 제33권5호
• /
• pp.397-405
• /
• 2023

항산화제로서 산화적 손상의 방지에 중요한 역할을 하는 것으로 알려진 glutathione (GSH)의 면역 조절에 대한 연구는 현재까지 제대로 이루어지지 않았다. 본 연구에서 우리는 환원형 GSH인 luthione^®이 RAW 264.7 세포에서 면역 강화 효과가 있는지를 조사하였다. 유세포 분석 및 면역 형광 실험의 결과에 의하면, luthione은 대조군 세포에 비해 대식세포의 대표적인 기능인 식세포 활성을 luthione 처리 농도 의적으로 증가시키는 것으로 나타났다. 또한, cytokine array의 결과에 의하면, IL-5, IL-1β와 IL-27의 발현이 luthione이 처리된 세포에서 유의하게 증가하였다. 아울러 luthione에 의한 TNF-α 및 IL-1β의 생성 증가는 그들의 단백질 발현 증가를 통해 이루어졌으며, NO 및 PGE₂와 같은 면역 매개체 유리의 증가는 iNOS 및 COX-2의 발현 증가와 관련이 있었으며, 이는 M1 대식세포 분화 마커인 CD86 발현의 증가와 연관성이 있었다. 그리고 heatmap 분석을 통하여 SOCS1/3 매개 STAT/JAK 신호 전달 경로가 luthione에 의한 면역 조절 증가에 관여함을 확인하였다. 결론적으로, 우리의 결과는 luthione이 M1 macrophage polarization의 분자 조절자로 작용하여 면역 능력을 향상시킬 수 있음을 시사한다.

• 생약학회지
• /
• 제43권4호
• /
• pp.328-337
• /
• 2012

In order to investigate anti-inflammatory, anti-allergic and anti-obesity activities of Samchulkunbi-tang (SCT; Shen zhu jian pi-tang) water and 70% ethanol (EtOH) extracts, in vitro inhibitory activities against nitric oxide (NO), prostaglandin $E_2$ $PGE_2$), interleukin (IL)-6 and tumor necrosis factor (TNF)-${\alpha}$ production in lipopolysaccharide-stimulated RAW 264.7 cells, and macrophage-derived chemokine (MDC/CCL22) and regulated on activation of normal T-cell-expressed and -secreted (RANTES/CCL5) production in TNF-${\alpha}$/interferon-${\gamma}$-stimulated HaCaT and BEAS-2B cells as well as glycerol-3-phosphate dehydrogenase (GPDH) activity and leptin production in 3T3-L1 cells were determined. A HPLC was used for quantification of the seven marker components (albiflorin, paeoniflorin, liquiritin, naringin, hesperidin, poncirin and glycyrrhizin) of SCT water and 70% EtOH extracts. SCT showed inhibitory effects against MDC and RANTES production in HaCaT cells, as well as RANTES production in BEAS-2B cells. In addition, SCT reduced not only NO, $PGE_2$, IL-6 and TNF-${\alpha}$ production in RAW 264.7 cells, but also GPDH activity and leptin production in 3T3-L1 cells. Furthermore, the biological activities and the contents of six compounds (except paeoniflorin) were higher in 70% EtOH extract than water extract. These results suggest that SCT has anti-inflammatory, anti-allergic and anti-obesity activities. These efficacies of 70% EtOH extract are relatively higher than that of water extract.

• 한국식품위생안전성학회지
• /
• 제32권6호
• /
• pp.536-541
• /
• 2017

본 연구는 까마귀쪽나무열매추출물(LJF-HE)이 흰쥐모델에서 스트레스로 유발되어지는 위염에 대한 억제효과를 확인하고자 하였다. 이때 까마귀쪽나무열매추출물(LJF-HE)소재는 지표성분인 hamabiwalactone B의 함량이 $15.23{\pm}0.057mg/g$로 규격에 적합한 것을 사용하였다, 동물실험에 있어 군분리는 정상군(normal), 양성 대조군(control, 수침 구속 스트레스 위염 유발), 약물 대조군(ranitidine, 50 mg/kg), 까마귀쪽나무열매추출물 저농도 투여군(LJF-HE-L, 30 mg/kg), 까마귀쪽나무열매추출물 중농도 투여군(LJF-HE-M, 60 mg/kg), 까마귀쪽나무열매추출물 고농도 투여군(LJF-HE-H, 120 mg/kg)의 총 6군으로 구성하여 실험을 진행하였다. 그 결과 까마귀쪽나무열매추출물의 투여그룹(LJF-HE-L, LJF-HE-M, LJF-HE-H)에서 염증의 길이가 control 그룹에 비하여 통계적으로 유의하게 감소하였으며, 육안 병변 관찰에서도 까마귀쪽나무열매추출물(LJF-HE) 투여그룹에서의 위 염증과 점막출혈 부위가 control 그룹에 비하여 감소하였음을 관찰할 수 있었다. 또한 까마귀쪽나무열매추출물(LJF-HE) 투여그룹에서의 펩신 활성도도 control 대비 유의성 있게 감소하는 것으로 나타나 까마귀쪽나무 열매추출물은 펩신 활성도를 낮춰 위염 발생을 억제하는 것으로 사료된다. 그리고 까마귀쪽나무열매추출물의 투여 그룹(LJF-HE-M, LJF-HE-H)에서 gastrin에 의해 활성화 되는 CCK-2r 유전자 발현이 대조군에 비해 유의적으로 억제되는 것으로 나타났으며, 염증성 cytokine중에 하나인 IL-$1{\beta}$의 혈장 내 함량이 대조군에 비해 유의적으로 감소하였고, 세포보호물질로 점액 및 혈류량을 증가시켜 위점막을 보호하는 역할을 하는 PGE2의 혈장 내 함량이 대조군에 비해 유의적으로 증가한 결과를 얻었다. 이와 같은 결과는 까마귀쪽나무열매추출물(LJF-HE)이 스트레스로 유발되어지는 위염에 대한 억제효과가 있음을 확인하였다.

• Journal of Chest Surgery
• /
• 제21권3호
• /
• pp.425-446
• /
• 1988

After 24 hours of preservation under 15 mmHg perfusion pressure the recovery rates of isolated canine hearts were determined. Preservation was performed in a cold room maintained at 4*C with 4 different types of perfusates bubbled with a mixture of 95% 0y and 5% CO~ using a modified perfusion unit designed in our institute. The perfusates used were as follows; Group 1: Krebs-Henseleit solution, Group 2: Krebs solution added by albumin and PGE1. Group 3: Modified Wicomb*s solution, Group 4: Modified Collin*s solution. The extent of myocardial recovery was evaluated using a modified isolated carmine perfusion model by measuring heart rate, systolic arterial pressure, left atrial pressure[LAP] and cardiac output. In addition to the above hemodynamic parameters, biochemical and enzymatic assays from perfusates and electron microscopic changes of the myocardium were also studied. The results were as follows; 1] The heart recovery rates were 41.6%, 53.4% and 108.9% in groups 1, 2 and 3, respectively, and group 3 elicited the best result[p< 0.001]. The heart beat was never recovered in group 4. 2] Recovered systolic arterial pressures[mmHg] were 63.3% in group 1, 94.9% in group 2 and 94.3% in group 3. 3] LAPs[mmHg] were 20 in group 1, 13.5 in group 2 and 11.2 in group 3, which suggested that the best myocardial preservation was elicited in group 3[p< 0.05]. 4] Cardiac output, the sum of aortic stroke volume and coronary leakage, were 69.1% in group 2, and 90.7% in group 3, but these were not statistically significant[p=0.24]. No aortic stroke output was measured in group 1 and 4. 5] The degree of myocardial edema increase was 17.5` in group 1, 24.6% in group 2, 20.9% in group 3 and 55.3% in group 4. But there were no statistical differences in each group[p= 0.08]. 6] CPK-MB[U/L] levels were increased 750% and 332%[p< 0.05], glucose levels[mg/dl] 60.5% and 78.2% and SGOT[U/L] levels 523% and 333%, in groups 2 and 3, respectively. Biochemical and enzymatic assays could not be performed in group 1 and group 4, because of poor recovery of heart beat. 7] Electron microscopic findings in the myocardium of most groups revealed slight to moderate muscle cell and mitochondrial edema. But all these findings were within the limits of reversible change. From these above results, it is suggested that modified Wicomb*s solution seems to be the most useful physiologic salt solution for preservation of the heart. We propose that after further study and improvement, our portable continuous hypothermic perfusion system will contribute to the development of a better preservation method for donor hearts for human heart transplantation.

• 대한한의학회지
• /
• 제34권3호
• /
• pp.69-85
• /
• 2013

Objectives: This study investigated the anti-osteoarthritic effects of Kyejiinsam-tang (hereinafter referred to KIT) on the monosodium iodoacetate (MIA)-induced osteoarthritis rats. Methods: Anti-oxidative effects of KIT were measured by scavenging activities of DPPH, reactive oxygen species (ROS) and nitric oxide (NO). Scavenging activities of anti-oxidation in lipopolysaccharide (LPS)-treated RAW 264.7 cells were also measured for inhibitory effects against the production of inflammatory mediators (tumor necrosis factor-${\alpha}$, interleukin-$1{\beta}$, interleukin-6). Osteoarthritis was induced in rats by injecting MIA in the knee joint. Rats were divided into a total of 4 groups (n=6). The normal group were not treated at all without inducing osteoarthritis whereas the control group were induced for osteoarthritis by MIA and oral medicated physiological saline per day. The positive comparison group was injected with MIA and after 7 days, 2 mg/kg of Indomethacin. The experimental group was injected with MIA and after 7 days was medicated with 34 mg/kg of KIT. Indomethacin and KIT were orally-medicated for each substance a total of 4 weeks, once per day. Weight-bearing on hind legs was measured every week after MIA injection. At the end of the experiment (5 weeks after MIA injection), micro CT (computed tomography)-arthrography and histopathological examinations on the articular structures of knee joint were performed. The effect on inflammatory cytokines and immunological cells in synovial fluid was measured. Volume of cartilage was measured by micro CT-arthrography. Injury to synovial tissue was measured by H & E (hematoxylin and eosin), Safranin-O immunofluorescence. Results: 1. Cytotoxicity against hFCs was insignificant. 2. KIT showed the potent full term for DPPH. 1. NO was significantly reduced by KIT (at 100, $200{\mu}g/m{\ell}$) and ROS was also reduced, but not significantly, by KIT (at $200{\mu}g/m{\ell}$). 2. IL-6 and IL-$1{\beta}$ were significantly reduced by KIT (at 100, $200{\mu}g/m{\ell}$) and TNF-${\alpha}$ was also reduced, but not significantly, by KIT (at $200{\mu}g/m{\ell}$). 1. In hind legs weight-bearing measurement, level of weight increased. 2. Functions of liver and kidney were not affected. 3. IL-$1{\beta}$ was significantly reduced and TNF-${\alpha}$, IL-6 were also reduced but not significantly. 4. PGE2 (prostaglandin E2), LTB4 (leukotriene B4) were significantly reduced in the KIT group. 5. MMP-9 (matrix metalloproteinase-9), TIMP-1 (tissue inhibitor of metalloproteinases-1) and Osteocalcin were significantly reduced in the KIT group. 6. Destruction of cartilage on micro CT arthrography was reduced but had no significant differences. 7. Histopathologically, injury to synovial membrane of the KIT group was decreased and proteoglycan content of KIT group was increased. Conclusions: According to this study, Kyejiinsam-tang has inhibiting effect on the progression of arthritis in MIA-induced osteoarthritis rat. Kyejiinsam-tang has anti-oxidants and anti-inflammation effects, and is related to inhibiting the activity of inflammatory cytokine and injury of volume in cartilage.

• Molecules and Cells
• /
• 제40권3호
• /
• pp.222-229
• /
• 2017

Adipose-derived stem cells (ADSCs) were previously considered to have an anti-inflammatory effect, and Interleukin-$1{\beta}$ ($IL-1{\beta}$) was found to be a pro-inflammatory factor in chondrocytes, but the mechanism underlying ADSCs and $IL-1{\beta}$ is unclear. In this study, we investigate whether P2X7 receptor (P2X7R) signalling, regulated by microRNA 373 (miR-373), was involved in the ADSCs and $IL-1{\beta}$ mediated inflammation in osteoarthritis (OA). Chondrocytes were collected from 20 OA patients and 20 control participants, and ADSCs were collected from patients who had undergone abdominal surgery. The typical surface molecules of ASDCs were detected by flow cytometry. The level of nitric oxide (NO) was determined by Griess reagent. Concentrations of prostaglandin E2 (PGE2), interleukin 6 (IL-6), matrix metallopeptidase 3 (MMP-3) were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of IL-6, MMP-3, miR-373 and P2X7R were determined by real-time polymerase chain reaction (PCR), and Western blot was used to detect the protein expression of P2X7R. The typical potential characters of ADSCs were verified. In chondrocytes or OA tissues, the miR-373 expression level was decreased, but the P2X7R expression was increased. $IL-1{\beta}$ stimulation increased the level of inflammatory factors in OA chondrocytes, and ADSCs co-cultured with $IL-1{\beta}$-stimulated chondrocytes decreased the inflammation. OA chondrocytes transfected with the miR-373 inhibitor increased the inflammation level. The miR-373 mimic suppressed the inflammation by targeting P2X7R and regulated its expression, while its effect was reversed by overexpression of P2X7R. $IL-1{\beta}$ induced inflammation in OA chondrocytes, while ADSCs seemed to inhibit the expression of P2X7R that was regulated by miR-373 and involved in the anti-inflammatory process in OA.

• 생약학회지
• /
• 제43권1호
• /
• pp.39-45
• /
• 2012

Cirsium japonicum var. ussuriense has long been used in herbal medicine for the treatment of arthritis, dyspepsia, and bleeding in Korea. In the present study, we investigated anti-inflammatory effects of C. japonicum var. ussuriense against lipopolysaccharide(LPS)-induced activation in RAW264.7 cells by the expression of heme oxygenase (HO)-1. The 70% EtOH extract of the aerial parts of C. japonicum var. ussuriense (CJE), showed the potent anti-inflammatory effects on LPS-induced inflammation in RAW264.7 cells. The anti-inflammatory effect of CJE was demonstrated by the suppression of pro-inflammatory mediators, including pro-inflammatory enzymes (inducible nitric oxide synthase and cyclooxygenase-2). Furthermore CJE induced HO-1 expression through nuclear translocation of nuclear factor E2-related factor 2 (Nrf2) and increased HO activity in RAW264.7 macrophages. The effects of CJE on LPS-induced NO and $PGE_2$ productions were partially reversed by an HO-1 inhibitor, tin protoporphyrin (SnPP). Therefore, it is suggested that CJE-induced HO-1 expression plays a role of the resulting anti-inflammatory effects in macrophages. These results suggest that CJE may be a promising candidate for the treatment of inflammatory diseases.

• 동의생리병리학회지
• /
• 제33권2호
• /
• pp.131-140
• /
• 2019

The objective of present study is to evaluate concentration-dependent in vitro anti-osteoarthritic (OA) effects of synergic mixed formula consisted of dried pomegranate juice concentrate powder, Eucommiae Cortex aqueous extract and Achyranthis Radix aqueous extract 5:4:1 (g/g) mixture on the primary cultured rat articular chondrocytes. First, any cytotoxic effect of mixture was observed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium Bromide) assay. Next, cyto-protective effect of test substances was evaluated by using the recombinant human interleukin $(rhIL)-1{\alpha}$ induced chondrocytes. In addition, anti-inflammatory effects were also observed on the lipopolysaccaride (LPS) treated chondrocytes through prostaglandin $E_2(PGE_2)$ productions and 5-lipoxygenase (LPO) activities, and inhibitory effects on matrix metalloproteinase (MMP)-2 and MMP-9 activities were observed on $rhIL-1{\alpha}$ treated chondrocytes with their extracellular matrix (ECM) related mRNA expressions. No obvious cytotoxic effects of mixture were demonstrated. Inflammatory damages of chondrocytes and related ECM degradations induced by treatment of LPS or $rhIL-1{\alpha}$ were significantly and concentration-dependently inhibited by pretreatment of mixture from a concentration level of 0.001 mg/ml to 1 mg/ml. In addition, mixture showed $IC_{50}$ for $rhIL-1{\alpha}-induced$ MMP-2 and MMP-9 activities as 44.01 and $162.47{\mu}g/ml$, and also showed $EC_{50}$ for $rhIL-1{\alpha}-induced$ inhibition of collagen type II, SOX9 and aggrecan mRNA expression as 8.61, 10.79 and $4.47{\mu}g/ml$, respectively. It is observed that mixture showed concentration-dependent anti-inflammatory and cytoprotective ECM preserved effects on the primary cultured rat articular chondrocytes without cytotoxicity.

• Journal of Ginseng Research
• /
• 제13권2호
• /
• pp.202-210
• /
• 1989

인삼 성분이 prostaglandin 등 arachidonic acid 대사산물 생성에 미치는 영향을 실험함으로써 인삼의 약리학적 작용기전을 간접적으로 모색하였다. 즉, [$^3H$]-arachidonic acid를 기질로 넣어주고 토끼 신장 micorsome, 소의 대동맥 microsome, 정상 성인의 혈소판 homogenate 등을 효소원으로 한 in vitro 생합성 과정에 변화를 주는 수종 인삼 saponin 성분의 효과를 검정하였다. 실험에 사용한 인삼 saponin 성분은 panaxadiol, panaxatriol 및 protopanaxadiol계 soponin류인 Ginsenoside $Rb_2$(G-$Rb_2$), Ginsenoside Rc(G-Rc) 및 protopanaxatriol계 saponi류인 Ginsenoside (G-$Rb_2$)이었다. 1. Arachidonic acid로부터 생성된 총 cycoloxygenase 반응생성물 및 malondialdehyde의 양은 실험에 사용한 인삼 saponin 성분의 전 농도 범위에서 유의적인 변화를 보이지 않았는데 이는 인삼 saponin 성분들은 cyclooxygenase에 직접 작용하지 않는다는 것을 설명해 준다. 2. Panaxdiol($500{\mu}g$/ml)은 $PGE_2$ 생성에는 영향이 없으나 $PGF_2$ 및 $TXB_2$의 생성을 감소시켰으며 동시에 6-keto-$PGF_{1{\alpha}}$의 생성은 증가시켰다. Panaxatriol도 유사한 양상을 보였다. 3. G-$Rb_2$, Rc, Re에 의해 $TXB_2$의 생성은 농도 의존적으로 감소하였으나 6-keto-$PGF_{1{\alpha}}$의 생성은 유의적으로 증가하였다. 또한 arachidonic acid와 $TXA_2$ 유사제인 U46619(9,11-methanoepoxy $PGH_2$)로 유도한 혈소판 응집 현상은 세 ginsenoside에 의해 억제되었다. G-Re의 6-keto-$PGF_{1{\alpha}}$생성증가 효과는 prostacyclin 합성효소억제제에 의해 길항하였다. 이상의 결과와 같이 인삼saponin 성분들은 arachidonic acid로부터 cyclooxygenase를 통해 일단 생성된 endoperoxide에서 각각의 prostaglandin을 생성하는 효소, 특히 G-$Rb_2$는 $TXA_2$ synthetase에 강력한 억제제로, G-Re는 prostacyclin 생합성에 촉진데로 심혈관계 균형에 기여하리라 생각된다.

• 대한한의학회지
• /
• 제34권1호
• /
• pp.116-135
• /
• 2013

Objectives: This study was intended to clarify how Jakyakkamchobuja-tang (hereinafter referred to JKBT) affects mice of C57BL/10 whose osteoarthritis was induced by papain. Methods: Osteoarthritis was induced in mice by injecting papain in the knee joint. Mice were divided into 4 groups (n=6). The normal group were not treated at all whereas the control group (OAC-control) were induced for osteoarthritis by papain and oral medicated with 200 ul of physiological saline per day. The positive comparison group (OAC-$Joins^{(R)}$) were injected with papain and after 7 days, 100 mg/kg of $Joins^{(R)}$ were medicated with 200 ul of physiological saline mixed. The experimental group (OAC-JKBT) were injected with papain and after 7 days were medicated with 400 mg/kg of JKBT mixed with 200 ul of physiological saline. OAC-$Joins^{(R)}$ and OAC-JKBT were oral medicated for each substance for a total of 4 weeks, once per day. After experiments (from 1 week after injection of papain to 4 weeks elapsed), the function of liver and kidney, inflammation cytokine values within serum, degree of revelation for inflammation cytokine genes, immune cells within blood, metabolism of arachidonic acid and amount of cartilage were measured and histopathological variations for knee joint structures were observed. Results: Functions of liver and kidney were not affected. IL-$1{\beta}$ (interleukin-$1{\beta}$), MCP-1 (monocyte chemoattractant protein-1) and TNF-${\alpha}$ (tumor necrosis factor-${\alpha}$) were significantly reduced and IL-6 (interleukin-6) was also reduced but not significantly. After analyzing inflammation cytokine in joints with mRNA (messenger ribonucleic acid), revelation of IL-6, TNF-${\alpha}$, COX-2 (cyclooxygenase-2) and iNOS-II (inducible nitric oxide synthase-II) were all significantly reduced. Revelation of IL-$1{\beta}$ gene was also reduced but not significantly. Neutrophil for WBC (white blood cell) within serum was significantly reduced; monocyte was also reduced but not significantly. PGE2 (prostaglandin E2), TXB2 (thromboxane B2) were significantly reduced and LTB4 (leukotriene B4) was also reduced but not significantly. Destruction of cartilage on micro CT (computed tomography)-arthrography was reduced but had no significant differences. In terms of histopathology, infiltration of inflammation, proliferation of synovial membrane, subsidence of cartilage and bone due to penetration of excessive formation of synovial cell and destruction of cartilage were small (H&E (hematoxylin and eosin), safranine O staining). Conclusions: Based on these results, Jakyakkamchobuja-tang (JKBT) is believed to be useful for suppressing the progress of osteoarthritis and its treatments because of its anti-inflammatory effects and alleviation of pain with histopathological effective efficacy.