• 생명과학회지
• /
• 제30권1호
• /
• pp.106-112
• /
• 2020

아미노아실-트랜스퍼 리보핵산 합성효소-상호작용 다기능 단백질 2(AIMP2)는 여러 tRNA 합성효소들과의 결합체를 이루게 하는 기능을 하며, DNA 손상에 대한 반응으로 세포사멸 활성을 나타낼 수 있다. DNA에 손상이 발생하면 AIMP2는 MDM2 공격으로부터 p53을 보호하기 위해 MDM2에 결합한다. TGF-β 신호에서 AIMP2는 세포 핵으로 들어가 FUSE 결합 단백질(FBP)과 결합하여 c-myc을 억제한다. TNF 수용체 관련 인자 2(TRAF2)는 c-Jun N-말단 키나아제(JNK), NF-κB 및 p38 미토겐 활성화 단백질 키나아제(MAPKs)의 신호에서 실행되는 두 수용체, TNF 수용체 1과 2 사이의 중요한 중재자이다. TARF2는 TNF-α 신호에서 JNK와 NF-κB의 활성화에 필요하며, 세포사멸 신호를 막는 중재자 역할을 수행한다. 또한 TNF-α 신호에서 AIMP2는 세포사멸을 향상시킨다. 이 신호에서, AIMP2는 TRAF2를 분해하는 것으로 잘 알려진 E3 유비키틴 효소인 c-IAP1과의 결합을 향상시킨다. AIMP2, TRAF2 및 c-IAP1을 포함한 복합체의 형성은 proteasome을 매개로 하여 TRAF2의 분해를 초래한다. 이러한 연구 결과는 AIMP2가 TNF-α 신호에서 직접적인 상호작용을 통해 TRAF2를 하향 조절시켜 세포사멸을 유도할 수 있음을 시사한다.

• Clinical and Experimental Reproductive Medicine
• /
• 제28권2호
• /
• pp.141-145
• /
• 2001

Objective: We inversigated Small Heterodimer Partner (SHP) gene mutation in Korean Polycystic Ovarian Syndrome (PCOS) patients. SHP protein regulates the activity of nuclear receptors which regulate the cellular development and differentiation. Recently, the mutation of SHP gene was found in the obesity and diabetes patients in Japanese group, and suggested that its mutation may involved in pathogenic mechanism of PCOS. Methods: This study was performed in 20 PCOS patients and 20 normal women. The DNAs were extracted from the peripheral bloods, and amplified at each exon (1 and 2) of SHP gene by PCR method. Subsequently, each PCR product was digested with the restriction enzyme indicated below for studying restriction fragment length polymorphism (RFLP). After enzyme digestion, the results of RFLP were compared PCOS patients with control women to find any sequence variation. Results: We examined 9 regions of exon 1 with Msp I, Pvu II, Dde I and 3 regions of exon 2 with Pst I, Dde I. There is no heterozygous or homozygous mutation in patients and control women at these restriction sites. Conclusion: The genetic analysis at our restriction sites in the SHP gene did not show any genetic variation in Korean PCOS patients. Our PCR-RFLP analysis was not covered the entire SHP gene (68 bp/1,006 bp), we need to further analysis of the entire SHP gene.

• 동의생리병리학회지
• /
• 제26권3호
• /
• pp.320-324
• /
• 2012

Osteoporosis is an important public health issue in postmenopausal women. It is a major public health concern and is widely believed that osteoporosis results from imbalance between bone resorption and bone formation. Recently natural products from plants have been extensively studied as therapeutic drugs to treat and prevent various diseases. Hoelen (scientific name, Poria cocos) is a mushroom that is used in traditional Chinese medicine. Hoelen exhibits anti-inflammatory activity and has a protective effect on tumor progression. However, the effect of hoelen in osteoclast differentiation remains unknown. Thus, we examined the effect of hoelen in receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL)-induced osteoclast differentiation. Hoelen significantly inhibited RANKL-induced osteoclast differentiation in bone marrow-derived macrophages (BMMs) in dose dependent manner without toxicity. Also, we showed that hoelen significantly inhibited the mRNA expression of tartrate-resistant acid phophatase (TRAP) and nuclear factor of activated T cells 1 (NFATc1) in BMMs treated with RANKL. In Particular, hoelen greatly inhibited the protein expression of NFATc1. Ectopic expression of NFATc1 partially reverses hoelen-mediated inhibition of osteoclast differentiation. Taken together, our results demonstrated that hoelen may be useful treatment option of bone-related disease such as osteoporosis, reumatoid arthritis, and periodontitis.

• 대한핵의학회지
• /
• 제36권2호
• /
• pp.133-139
• /
• 2002

목적: 섬광 근접측정법은 항원 항체 반응 후 결합 분획과 유리분획을 분리하는 과정이 필요없다. 이러한 원리를 검체 내 hCG와 항 ${\alpha}$ hCG 항체간의 항원 항체 반응에 적용하고자 한다. 대상 및 방법: 항 ${\alpha}$ hCG 항체를 biotin과 결합시켜 SPA bead에 부착된 streptavidin과 부착 가능하게 만들었다. 이 측정법은 항 ${\alpha}$ hCG 항체가 부착된 SPA beads에 대해 혈청내 hCG와 표지항원인 $[^{125}I]hCG$간의 경쟁 반응을 기본 원리로 이용하였다. Biotin 표지 항 ${\alpha}$ hCG 항체를 $[^{125}I]hCG\;100{\mu}{\ell}$와 표준용액이나 환자 혈청 $200{\mu}{\ell}$이 들어있는 실온에서 20분 방치하였다. 그리고 streptavidin이 붙은 SPA beads $20{\mu}{\ell}$를 바이알에 넣고 10분 더 방치한다. 환자 혈청의 수치를 표준 응답곡선을 통해 계산하였다. 결과: SPA측정법에 사용되는 방사성 핵종의 방사능 양에 따라 반응용액 속에서 SPA bead와 자유 방사성 핵종에 의한 배후 방사능이 측정값에 영향이 없음을 확인하였다. SPA 방법을 응용한 측정에서 적합한 표준 응답곡선을 얻었고, 실제 환자혈청에서의 hCG 농도를 결정할 수 있었다. 결론: 이 실험을 통해 SPA 방법을 이용한 측정법이 임상진단에 유용하게 사용될 수 있을 것으로 사료된다.

• Journal of Acupuncture Research
• /
• 제32권3호
• /
• pp.69-81
• /
• 2015

Purpose : The first purpose of this study is to find out whether the water extract of Rehmanniae Radix Preparat(RRP), Cuscutae Semen(CS) and their combination(Ssangbohwan, SBH) have the effect of suppressing Receptor activator of nuclear factor kappa-B ligand(RANKL)-induced osteoclast differentiation. The second purpose of this study is to find out whether the water extract of RRP, CS and SBH have the effect of inhibiting osteoporosis in an osteoporosis model induced by lipopolysaccharide(LPS). Methods : After promoting differentiation of osteoclasts by treating the RANKL, we observed the effect by the administration of RRP, CS and SBH. In addition, by means of Reverse transcription polymerase chain reaction(RT-PCR), we assayed mRNA expression levels of NFATc1, c-Fos, TRAP and GAPDHS(Glyceraldehyde-3-phosphate dehydrogenase, spermatogeni) from bone marrow macrophages(BMMs). Similarly, the protein expression levels of NFATc1 (Nuclear factor of activated T-cells, cytoplasmic1), C-Fos, MAPKs(Mitogen-activated protein kinases) and ${\beta}$-actin in cell lysates were analyzed by means of Western Blotting. Finally, we determined the anti-osteoporotic effects of RRP, CS and SBH, through the use of Lipopolysaccharide-induced bone-loss mouse. Results : RRP, CS and SBH showed remarkable inhibitive effect on RANKL-treated osteoclast differentiation without cytotoxicity. SBH inhibited the phosphorylation of p38, Jun N-terminal kinases(JNK), and I-${\kappa}B$ and down-regulated the induction of c-Fos and NFATc1 by RANKL. RRP, CS suppressed degradation of I-${\kappa}B$, but it did not affect c-Fos and NFATc1 by RANKL. Lastly, in vivo data showed that RRP and SBH prevented bone erosion by LPS treatment. Conclusions : These results demonstrate SBH can be effective remedy for bone-loss diseases such as osteoporosis.

• Natural Product Sciences
• /
• 제26권1호
• /
• pp.83-89
• /
• 2020

Osteoporosis is a worldwide disease leading to significant economic and societal burdens globally. Osteoporosis is caused by unbalanced bone remodeling between the rate of osteoclast bone resorption and osteoblast bone formation. Acer tegmentosum Maxim (AT) is a traditional herbal medicine containing multiple biological activities such as anti-oxidant and anti-inflammatory purposes. However, its role in osteoporosis has not been fully studied. Therefore, we investigated whether AT has a potent inhibitory effect on osteoporosis and its mechanism through a systemic evaluation in ovariectomized (OVX) mice. OVX mice were orally administrated with the AT at doses of 50, 100, and 200 mg/kg for 10 weeks. Histological images and histomorphometry analyses were performed by H&E and Toluidine blue satin, and the expression levels of receptor activator for nuclear factor-kB ligand (RANKL), nuclear factor of activated T cells cytoplasm 1 (NFATc1), c-Fos, and matrix metalloproteinase 9 (MMP9) related to the osteoclast differentiation were investigated using immunohistochemical analysis. Administration of AT prevented bone loss and the alternations of osteoporotic bone parameters at the distinct regions of the distal femur and spongiosa region in OVX mice. Further, administration of AT increased periosteal bone formation in a dose-dependent manner. Meanwhile, AT inhibited not only the expression of NFATc1 and c-Fos, which are two major regulators of osteoclastogenesis but also reduced bone resorbed encoding expression of MMP9 and RANKL. Our results indicated that administration of AT prevented bone loss and the alternations of osteoporotic bone parameters at the distinct regions of the distal femur and spongiosa region in OVX mice. Also AT has the bone protective effect through the suppression of osteoclast and promotion of osteoblast, suggesting that it could be a preventive and therapeutic candidate for anti-osteoporosis.

• 생명과학회지
• /
• 제22권11호
• /
• pp.1571-1586
• /
• 2012

지구상에는 약 5,400여 종의 포유동물이 있고 그 중 약 1,000여 종은 풀을 뜯어 먹고 사는 초식동물이다. 초식동물 중에서 약 250여 종이 반추동물로 알려져 있다. 반추동물인 소와 양은 반추위에서 주로 발효가 일어나지만 비반추동물인 돼지와 사람은 맹장과 결장, 직장에서 주로 발효가 일어난다. 반추위 미생물의 종류와 우점도 Bacteroidetes 51%, Firmicutes 43% 존재하며, 사람의 대장미생물의 우점도Firmicutes 65%, Bacteroidetes 25%로 존재한다. 풀의 세포벽 구성성분은 미생물에 의해 분해, 발효에 의해 SCFA (short chain fatty acid)를 생성하게 되고 acetate, propionate, butyrate 생성비율은 60:25:15이다. 장내 primary butyrate transporter인 MCT1(monocarboxylatetransports-1)에 의해서 흡수된 butyrate는 SCFA receptor GPR43과 GPR41을 활성화시킨다. Butyrate는 강력한 anti-tumorigenic 기능을 가지고 있다. Butyrate는 다양한 cancer cell에 효과를 나타내며 세포내의 기능 조절에 기여하고, 암세포사멸을 유도하는 특성이 있다. Butyrate는 caspase의 활성화, HDAC (histone deacetylase) 활성을 억제하여apoptosis를 유도하고, p53 발현증가로 cell cycle arrest와 apoptosis를 유도한다. SCFA의 항 염증작용으로는 장 상피세포에서 IL-8 발현 감소, NO합성과 NF-${\kappa}B$ (nuclear factor ${\kappa}B$)의 활성을 억제하여 염증으로 인한 암 발생을 억제한다. Butyrate는 장 점막의 생리적 기능을 유지하는데 중요한 역할을 하며 IBD (inflammatory bowel disease) 치료법으로 이용되고 있다.

• 한국식품영양학회지
• /
• 제29권2호
• /
• pp.178-185
• /
• 2016

This study was designed to develop and to qualify a coffee alternative beverage using a mixture of coffee beans and roasted black beans (Rhynchosia nulubilis). Therefore, the total isoflavone content (TIC), total phenol content (TPC), antioxidant activity, anti-inflammatory activity, NFATc1 (Nuclear factor of activated T-cells c1) expression in RANKL (receptor activator of nuclear factor kappa-B ligand)-stimulated RAW264.7 cells and sensory evaluation were measured for 5 different Cb (coffee bean)-RoS (roasted seomoktae) mixture extracts (Cb100RoS0, Cb75RoS25, Cb50RoS50, Cb25RoS75, and Cb0RoS100). Cb0RoS100 had the highest TIC ($516.83{\pm}36.61mg/100g$) and TPC ($18.11{\pm}1.77mg$ TAE/100 g) along with the highest antioxidant activity as measured by DPPH radical scavenging activity ($73.55{\pm}8.11%$) and ABTS radical scavenging activity ($63.27{\pm}7.27%$). Also, Cb0RoS100 showed the highest anti-inflammatory activity as measured by NO production ($13.57{\pm}2.21{\mu}M$) and PGE2 production ($3.25{\pm}0.21ng/mL$). The more the RoS ratio was increased in the mixtures of Cb-RoS, the more the NFATc1 protein expression was decreased in RANKL-stimulated RAW264.7 cells. In case of sensory evaluation, Cb50RoS50 had the highest scores for flavor, delicate flavor and overall quality, which were similar to those in Cb alone (Cb100RoS0). We suggest that the use of RoS replacement instead of Cb in/as a coffee alternative beverage may help to reduce the risk of caffeine-related bone loss and/or bone disease by effectively blocking NFATc1 expression in RANKL-stimulated RAW264.7 cells compared with Cb alone.

• 대한기관식도과학회지
• /
• 제16권2호
• /
• pp.109-114
• /
• 2010

Background: The aim of this study was to compare a novel mannose receptor-binding agent(Technetium-99m human serum albumin, $^{99m}Tc$-MSA) with $^{99m}Tc$-phytate for sentinel node mapping in patients with esophageal cancer. Material and Method: Twenty patients with clinical stage T1N0m0 or T2N0M0 esophageal cancer that were candidates for esophagectomy were enrolled. Endoscopic injection of $^{99m}Tc$-MSA or $^{99m}Tc$-phytate was administered at the peri-tumor region before surgery in 10 patients. The radioactive lymph nodes were identified with a handheld gamma probe after lymph node dissection. Results: The patient's age and the sex ratio of both groups were similar. The clinical stage, tumor location, and operative technique did not differ. The total number of dissected lymph nodes did not differ ($28.5{\pm}9.12$ in MSA group, $32.2{\pm}11.24$ in phytate group, p=0.430). The sentinel node was identified in all cases in both groups. The number of sentinel nodes per patient was $2.7{\pm}1.57$ in the MSA group, which was significantly greater than the $1.7{\pm}0.88$ in the phytate group (p=0.036). Five out of 20 patients whose sentinel nodes could be identified had metastases; however, neither group had any false-negative results for sentinel node identification. Conclusion: Sentinel nodes were detected more frequently with MSA than with phytate.

• 한국자원식물학회:학술대회논문집
• /
• 한국자원식물학회 2019년도 추계학술대회
• /
• pp.92-92
• /
• 2019

Excessive or chronic inflammation contributes to the pathogenesis of many inflammatory diseases such as sepsis, rheumatoid arthritis, and ulcerative colitis. Hibiscus syriacus L. has been used as a medicinal plant in many Asian countries, even though its anti-inflammatory activity has been unclear. Therefore, we investigated the anti-inflammatory effect of anthocyanin fractions from the H. syriacus L. varieties Pulsae (PS) on the lipopolysaccharide (LPS)-induced expression of proinflammatory mediators and cytokines in RAW264.7 macrophages. PS suppressed LPS-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) secretion concomitant with downregulation of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. Furthermore, PS inhibited the production of proinflammatory cytokines such as tumor necrosis factor-alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), and IL-12 in LPS-stimulated RAW264.7 macrophages. Further study showed that PS significantly decreased LPS-induced nuclear translocation of the nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) subunits, p65 and p50. Molecular docking data showed that many anthocyanins from PS fit into the hydrophobic pocket of MD2 and bound to Toll-like receptor 4 (TLR4), indicating that PS inhibits the TLR4-MD2-mediated inflammatory signaling pathway. Especially, apigenin-7-O-glucoside most powerfully bound to MD2 and TLR4 through LYS122, LYS122, and SER127 at a distance of $2.205{\AA}$, $3.098{\AA}$, and $2.844{\AA}$ and SER441 at a distance of $2.873{\AA}$ (docking score: -8.4) through hydrogen bonding, respectively. Additionally, PS inhibited LPS-induced TLR4 dimerization/expression on the cell surface, which consequently decreased MyD88 recruitment and IRAK4 phosphorylation. PS completely blocked LPS-mediated mortality in zebrafish larvae by diminishing the recruitment of neutrophil and macrophages accompanied by low levels of proinflammatory cytokines. Taken together, our results indicate that PS attenuates LPS-mediated inflammation in both in vitro and in vivo by blocking the TLR4/MD2-MyD88/IRAK4-$NF-{\kappa}B$ axis. Therefore, PS might be used as a novel modulatory candidate for effective treatment of LPS-mediated inflammatory diseases.