• Journal of Ginseng Research
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• 제46권1호
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• pp.156-166
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• 2022

Background: Panax ginseng Meyer (P. ginseng), a herb distributed in Korea, China and Japan, exerts benefits on diverse inflammatory conditions. However, the underlying mechanism and active ingredients remains largely unclear. Herein, we aimed to explore the active ingredients of P. ginseng against inflammation and elucidate underlying mechanisms. Methods: Inflammation model was constructed by lipopolysaccharide (LPS) in C57BL/6 mice and RAW264.7 macrophages. Molecular docking, molecular dynamics, surface plasmon resonance imaging (SPRi) and immunofluorescence were utilized to predict active component. Results: P. ginseng significantly inhibited LPS-induced lung injury and the expression of proinflammatory factors, including TNF-α, IL-6 and IL-1β. Additionally, P. ginseng blocked fluorescencelabeled LPS (LPS488) binding to the membranes of RAW264.7 macrophages, the phosphorylation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs). Furthermore, molecular docking demonstrated that ginsenoside Ro (GRo) docked into the LPS binding site of toll like receptor 4 (TLR4)/myeloid differentiation factor 2 (MD2) complex. Molecular dynamic simulations showed that the MD2-GRo binding conformation was stable. SPRi demonstrated an excellent interaction between TLR4/ MD2 complex and GRo (K_D value of 1.16 × 10^-9 M). GRo significantly inhibited LPS488 binding to cell membranes. Further studies showed that GRo markedly suppressed LPS-triggered lung injury, the transcription and secretion levels of TNF-α, IL-6 and IL-1β. Moreover, the phosphorylation of NF-κB and MAPKs as well as the p65 subunit nuclear translocation were inhibited by GRo dose-dependently. Conclusion: Our results suggest that GRo exerts anti-inflammation actions by direct inhibition of TLR4 signaling pathway.

• Biomolecules & Therapeutics
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• 제30권4호
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• pp.340-347
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• 2022

Advanced or metastatic breast cancer affects multiple organs and is a leading cause of cancer-related death. Cancer metastasis is associated with epithelial-mesenchymal metastasis (EMT). However, the specific signals that induce and regulate EMT in carcinoma cells remain unclear. PRR16/Largen is a cell size regulator that is independent of mTOR and Hippo signalling pathways. However, little is known about the role PRR16 plays in the EMT process. We found that the expression of PRR16 was increased in mesenchymal breast cancer cell lines. PRR16 overexpression induced EMT in MCF7 breast cancer cells and enhances migration and invasion. To determine how PRR16 induces EMT, the binding proteins for PRR16 were screened, revealing that PRR16 binds to Abl interactor 2 (ABI2). We then investigated whether ABI2 is involved in EMT. Gene silencing of ABI2 induces EMT, leading to enhanced migration and invasion. ABI2 is a gene that codes for a protein that interacts with ABL proto-oncogene 1 (ABL1) kinase. Therefore, we investigated whether the change in ABI2 expression affected the activation of ABL1 kinase. The knockdown of ABI2 and PRR16 overexpression increased the phosphorylation of Y412 in ABL1 kinase. Our results suggest that PRR16 may be involved in EMT by binding to ABI2 and interfering with its inhibition of ABL1 kinase. This indicates that ABL1 kinase inhibitors may be potential therapeutic agents for the treatment of PRR16-related breast cancer.

• Pediatric Infection and Vaccine
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• 제29권2호
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• pp.70-76
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• 2022

코로나바이러스감염증-19 (COVID-19)의 임상 양상은 무증상부터 급성 호흡곤란 증후군에 이르기까지 다양하다. 점액 다당류증(mucopolysaccharidosis) 2형은글라이코스아미노글라이칸(glycosaminoglycan)의 일종인 헤파란 황산염(heparan sulfate)과 더마탄 황산염(dermatan sulfate)의 분해를 촉매하는 효소 결핍에 의해 상기 물질이 리소좀(lysosome)에 축적되는 질환으로 전신 침범, 특히 호흡기침범을 특징으로 한다. 따라서 박테리아나 바이러스에 의한 호흡기 감염은 예후에 치명적일 수 있다. 현재 점액 다당류증 환자에서 제 2형 중증급성호흡기증후군 코로나 바이러스(SARS-CoV-2) 감염 후의 임상 양상에 대한 보고는 매우 드물고, 이에 점액 다당류증 2형으로 효소대체요법을 받고 있던 환자에서 상기 바이러스 감염 후의 임상 양상에 대해 보고하고 관련 문헌에 대해 고찰하고자 한다. 16세 남아로 가족간 전파로 코로나바이러스감염증이 발생하였다. 콧물, 기침, 가래 등 호흡기 증상이 관찰되었다. 발열이나 산소요구도 증가는 없었으며 심박수, 호흡수, 산소 포화도는 정상 범위였고 혈액검사결과에서 백혈구 감소증이 관찰되었다. 흉부 방사선 사진에서 폐렴 소견은 보이지 않았다. 보존적 치료와 격리만으로 증상이 호전되었다. 경미한 임상 양상의 원인으로 전구 물질의 축적으로 인해 바이러스에게 불리한 숙주의 세포 환경, 바이러스와의 상호작용에 관여하는 단백질을 암호화하는 유전자 발현의 특정방향으로의 변화가 제시되고 있다. 또한 점액 다당류증 환자에서 증가된 혈청 헤파란 황산염이 SARS-CoV-2 스파이크 단백질과 숙주 세포의 상호작용에 필수적인 세포 표면의 헤파란 황산염과 경쟁하여 SARS-CoV-2의 세포 내 침투로부터 보호한다는 가설도 있다. 향후 더 많은 사례를 통해 점액 다당류증 등의 리소좀 축적질환에서 코로나바이러스감염증의 발현 양상에 대한 연구가 필요하다.

• Journal of Animal Science and Technology
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• 제65권5호
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• pp.989-1001
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• 2023

The study evaluated the effects of dietary fiber and energy levels administered during two growing periods (d 0-28 and d 29-56) for pigs exposed to a high temperature. A total of 96 growing pigs were used in six treatments as: Two treatments in thermoneutral temperature (21℃-24℃) with dietary energy of 3,300 and the inclusion of high or low fiber, two treatments in heat stress (30℃-34℃) with dietary energy of 3,300 and the inclusion of high or low fiber, and two treatments in heat stress with dietary energy of 3,450 and the inclusion of high or low fiber. Among standard energy level treatments, heat-stressed pigs showed lower average daily gain (ADG), feed intake, digestibility of dry matter, gross energy, crude protein, and crude fiber in phases 1 and 2. Moreover, higher concentrations of acetate, propionate, butyrate, and total short-chain fatty acid (SCFA) in feces were shown in pigs fed high fiber diets. There was a negative interaction between dietary fiber and energy for the fecal concentration of isobutyrate in phase 1 and valerate in phase 2. Pigs in heat stress treatments showed a higher rectal temperature, respiratory rate, hair cortisol, plasma zonulin, and fecal lipocalin-2. Among heat stress treatments, the overall ADG was increased in pigs fed high fiber. Pigs fed high dietary fiber showed a greater concentration of acetate, propionate, butyrate, and total SCFA. High fiber treatments decreased plasma zonulin. In conclusion, the inclusion of beet pulp, soluble fiber, at the level of 4% looks necessary in pigs diet during heat stress.

• Journal of Ginseng Research
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• 제47권6호
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• pp.726-734
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• 2023

Background: Skeletal muscles play a key role in physical activity and energy metabolism. The loss of skeletal muscle mass can cause problems related to metabolism and physical activity. Studies are being conducted to prevent such diseases by increasing the mass and regeneration capacity of muscles. Ginsenoside Rg5 has been reported to exhibit a broad range of pharmacological activities. However, studies on the effects of Rg5 on muscle differentiation and growth are scarce. Methods: To investigate the effects of Rg5 on myogenesis, C2C12 myoblasts were induced to differentiate with Rg5, followed by immunoblotting, immunostaining, and qRT-PCR for myogenic markers and promyogenic signaling (p38MAPK). Immunoprecipitation confirmed that Rg5 increased the interaction between MyoD and E2A via p38MAPK. To investigate the effects of Rg5 on prevention of muscle mass loss, C2C12 myotubes were treated with dexamethasone to induce muscle atrophy. Immunoblotting, immunostaining, and qRT-PCR were performed for myogenic markers, Akt/mTOR signaling for protein synthesis, and atrophy-related genes (Atrogin-1 and MuRF1). Results: Rg5 promoted C2C12 myoblast differentiation through phosphorylation of p38MAPK and MyoD/E2A heterodimerization. Furthermore, Rg5 stimulated C2C12 myotube hypertrophy via phosphorylation of Akt/mTOR. Phosphorylation of Akt induces FoxO3a phosphorylation, which reduces the expression of Atrogin-1 and MuRF1. Conclusion: This study provides an understanding of how Rg5 promotes myogenesis and hypertrophy and prevents dexamethasone-induced muscle atrophy. The study is the first, to the best of our knowledge, to show that Rg5 promotes muscle regeneration and to suggest that Rg5 can be used for therapeutic intervention of muscle weakness and atrophy, including cancer cachexia.

• Clinical Nutrition Research
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• 제12권3호
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• pp.199-217
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• 2023

People with higher genetic predisposition to obesity are more susceptible to cardiovascular diseases (CVDs) and healthy plant-based foods may be associated with reduced risks of obesity and other metabolic markers. We investigated whether healthy plant-foods-rich dietary patterns might have inverse associations with cardiometabolic risk factors in participants at genetically elevated risk of obesity. For this cross-sectional study, 377 obese and overweight women were chosen from health centers in Tehran, Iran. We calculated a healthy plant-based diet index (h-PDI) in which healthy plant foods received positive scores, and unhealthy plant and animal foods received reversed scores. A genetic risk score (GRS) was developed based on 3 polymorphisms. The interaction between GRS and h-PDI on cardiometabolic traits was analyzed using a generalized linear model (GLM). We found significant interactions between GRS and h-PDI on body mass index (BMI) (p = 0.02), body fat mass (p = 0.04), and waist circumference (p = 0.056). There were significant gene-diet interactions for healthful plant-derived diets and BMI-GRS on high-sensitivity C-reactive protein (p = 0.03), aspartate aminotransferase (p = 0.04), alanine transaminase (p = 0.05), insulin (p = 0.04), and plasminogen activator inhibitor 1 (p = 0.002). Adherence to h-PDI was more strongly related to decreased levels of the aforementioned markers among participants in the second or top tertile of GRS than those with low GRS. These results highlight that following a plant-based dietary pattern considering genetics appears to be a protective factor against the risks of cardiometabolic abnormalities.

• Animal Bioscience
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• 제37권2호
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• pp.228-239
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• 2024

Objective: The objective of the experiment was to study yeast supplementation (yeast, Y) and dried kratom leaves (DKTL) on the digestibility, ruminal fermentation, blood metabolites and nitrogen balance in goats. Methods: Four of 7 to 8 months old male crossbred (50% Thai Native-Anglo Nubian) goats with average liveweight 20±0.13 kg were randomly assigned according to a 2×2 factorial arrangement in a 4×4 Latin square design to receive four diets ad libitum basis. The study investigated the effects of two levels of yeast (Y) supplementation (Y, 0 and 0.5g/kg dry matter [DM]) along with two levels of DKTL supplementation (DKTL, 0 and 4.44g/kg DM). The experimental groups were as follows: T1 = control group with 0Y+0DKTL, T2 = 0Y+4.44 DKTL, T3 = 0.5Y+0DKTL, and T4 = 0.5Y+4.44 DKTL. Results: The results showed that there were no interactions between Y levels and DKTL levels with respect to total DM intake, but there were significant effects (p<0.05) by levels of Y; goats receiving 0.05 g/kg DM Y had higher than goats fed 0.0 g/kg DM on average (kg/d). A percentage of body weight (% BW) and grams per kilogram of metallic weight (g/kg w^0.75) had no influence on yeast levels and DKTL, but there was a difference (p<0.05) by yeast level Y at 0.5 g/kg DM, being higher compared to the non-supplemented group. Apparent digestibility coefficient of nutrition in the form of (DM, organic matter, crude protein, neutral detergent fiber, and acid detergent fiber) was an increased trend in the Y-level complementary group at 0.5 g/kg DM and DKTL at 4.44 g/kg DM, respectively. Protozoa populations decreased in the group receiving Y levels at 0.5 g/kg DM and DKTL levels at 4.44 g/kg DM when compared to group T1. The acetic acid concentration and methane gas generation decreased (p<0.05) in the group receiving Y levels of 0.5 g/kg DM and DKTL levels of 4.44 g/kg DM, while the amount of propionic acid increased (p<0.05). Conclusion: Effects of feeding combinations of Y and DKTL supplementation on feed showed no interaction effect (Y×DKTL) on feed intake, rumen fermentation, bacterial and fungi population. The effect on protozoal populations was lower in the group that was supplemented with DKTL at 4.44 g/kg DM related to synthetic CH₄ was reduced.

• 대한수의학회지
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• 제15권1호
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• pp.57-67
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• 1975

The poxvirus group is considered to be a typical cytoplasmic inclusion forming virus. Every poxvirus has been reported to produce only one kind of inclusion in the infected tissues. A vague concept that inclusions of poxviruses are eosinophilic or acidophilic has prevailed. Although many papers and theories about the nature of the inclusion have been presented, most of them are not quite convincing on the point of the relations with virus multiplication, and an analysis of papers published showed that there seem to be many discrepancies in the descriptions of the nature of the poxvirus inclusions. Comparative studies on host-virus interaction with cowpox, orf, swinepox and fowlpox viruses which selected from each Group (I-IV) of poxviruses were performed from the morphological and virological standpoints. At first, in cowpox virus-FL cell system, as a comparative model, cytoplasmic inclusion, nucleic acid metabolism by autoradiography and detection of viral antigen by immunofluorescence were studied and obtained the results as follows: 1. The focus-like cytopathic effect (CPE) at early stage developed to entire culture at terminal stage of infection, and also the developing status of CPE was correlated to viral doses for inoculation. Two kinds of cytoplasmic inclusions which named A and B type were easily observed by Giemsa, hematoxylin-eosin (H & E) and May-Greenwald Giemsa (MGG) stainings in the infected cells. The B type inclusions were formed at early stage of infection and the A type inclusions were produced subsequently the B type formation. The B type which common type inclusion in poxviruses was a small compact or aggregate at early stage and developed to a large diffuse body at terminal stage of infection. On the other hand, the A type inclusion which depend upon the kind of virus was appeared as round and discrete shape, and its size and number was increased gradually during the culture period. It was characteristic to form distinct halos around the both types of inclusions in acid fixed, H & E stained preparations of infected cultures. The B type inclusion was always positive in Feulgen reaction and showed as DNA containing body but the A type inclusion was not. 2. In the relationship between inclusion and DNA metabolism of infected cells by the qualitative autoradiography using 3H-thymidine, the appearance of silver grains was coincided with B type inclusion but not with A type inclusion. This showed that the DNA synthesis was proceeded in all B type inclusions except those in the terminal stage with a diffuse form. This suggested that the B type inclusions are only sites of DNA synthesis and this was proceeded after the cell infection independently. The activity of DNA synthesis of the inclusions was nearly the same as that of the nucleic of normal cells and non-inclusion bearing cells. and non-inclusion bearing cells. Regardless of the size of the degree of DNA synthesis of the B type inclusion, inclusion bearing cells all showed remarkable suppression of nuclear DNA synthesis. 3. By the direct fluorescent antibody technique viral antigen in infected cells was detected. The B type inclusions have been proved to contain a great deal of viral antigen, whereas the basic substance of A type inclusion did not show antigenicity except the round edge. It was suggested that the round edge fluorescence might be caused by the glare of cytoplasmic viral antigen which pushed out and concentrated by the A type inclusion development. 4. Hemorrhagic red pock formations on chorioallantoic membrane of embryonated chicken egg had proved the characteristic of used viral strain. 5. By the above studies on the nature of two types of inclusions and the role they play in virus multiplication, it was concluded that the B type inclusion must be the site of the synthesis of viral DNA and protein as well as the site of the virus.

• IMMUNE NETWORK
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• 제3권1호
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• pp.1-7
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• 2003

Type II collagen (CII), major component of hyaline cartilage, has been considered as an auto-antigen in rheumatoid arthritis (RA). However, the clinical and biological significances with regard to the CII autoimmunity need to be clarified in human RA. The presence of antibodies to CII has been identified in sera, synovial fluid, and cartilage of patients with RA. In our study, the increased titer of IgG anti-CII in sera was well correlated with C-reactive protein, suggesting that this antibody may reflect the inflammatory status of RA. The titer of anti-CII antibodies (anti-CII Abs) tended to be higher in early stages of diseases. In our extending study, among 997 patients with RA, 269 (27.0%) were positive for circulatory IgG antibody to CII, those levels were fluctuated over time. It is hard to assess the significant amount of T cell responses to CII and CII (255~274) in RA. By using a sensitive method of antigen specific mixed lymphocyte culture, we can detect the presence of CII-reactive T cells in peripheral blood mononuclear cells of RA patients. Sixty seven (46.9%) of 143 patients showed positive CII reactive T cell responses to CII or CII (255~274). The frequencies of CII reactive T cells were more prominent in inflamed synovial fluid (SF) than in peripheral blood. These T cells could be clonally expanded after consecutive stimulation of CII with feeding of autologous irradiated antigen presenting cells (APC). Moreover, the production of Th1-related cytokine, such as IFN-${\gamma}$, was strongly up-regulated by CII reactive T cells. These data suggest that T cells responding to CII, which are probably presenting the IFN-${\gamma}$ producing cells, may play an important role in the perpetuation of inflammatory process in RA. To evaluate the effector function of CII reactive T cells, we investigated the effect of CII reactive T cells and fibroblasts-like synoviocytes (FLS) interaction on the production of pro-inflammatory cytokines. When the CII reactive T cells were co-cultured with FLS, the production of IL-15 and TNF-${\alpha}$ from FLS were significantly increased (2 to 3 fold increase) and this increase was clearly presented in accord to the expansion of CII reactive T cells. In addition, the production of IFN-${\gamma}$ and IL-17, T cell derived cytokines, were also increased by the co-incubation of CII reactive T cells with FLS. We also examined the impact of CII reactive T cells on chemokines production. When FLS were co-cultured with CII stimulated T cells, the production of IL-8, MCP-1, and MIP-1${\alpha}$ were significantly enhanced. The increased production of these chemokines was strongly correlated with increase the frequency of CII reactive T cells. Conclusively, immune response to CII was frequently found in RA. Activated T cells in response to CII contributed to increase the production of proinflammatory cytokines and chemokines, which were critical for inflammatory responses in RA. The interaction of CII-reactive T cells with FLS further augmented this phenomenon. Taken together, our recent studies have suggested that autoimmunity to CII could play a crucial role not only in the initiation but amplification/perpetuation of inflammatory process in human RA.

• 생명과학회지
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• 제30권2호
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• pp.137-146
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• 2020

식물에서 Ca²⁺는 세포의 중요한 2차 신호 전달 분자 중 하나이다. Ca²⁺ 및 인산화 효소의 센서 단백질인 칼슘-의존성 단백질 카이네즈(CDPKs)는 식물 세포에서 가장 풍부한 세린/트레오닌 키나아제이다. 이들은 다양한 자극에 대한 신호를 변환하여 식물에서 특정 반응을 일으킨다. 벼에는 31개의 CDPK 유전자 족이 확인되었다. 그들은 주로 식물의 생장과 발달에 관여하며 다양한 스트레스 조건에 반응하여 기능을 하는 것으로 알려져 있다. 그러나 CDPK 단백질의 생화학적 특성에 대해서는 알려진 바가 별로 없다. 이 연구에서는 벼의 CDPK 중 하나인 OsCPK11을 부분 정제하여 그 생화학적 특성을 조사하고자 하였다. 벼 유식물에서 3단계 칼럼 크로마토그래피 과정을 거쳐 부분 정제된 OsCPK11을 얻었다. 정제 과정에는 DEAE를 사용한 음이온 교환 크로마토그래피, Phenyl-Sepharose를 사용한 소수성 상호작용 크로마토그래피 및 Sephacryl-200HR를 사용한 겔 여과 크로마토그래피를 포함하였다. 부분 정제된 OsCPK11은 분자량이 54kDa이며 소수성 수지와 강한 소수성 상호작용을 보였다. 부분 정제된 OsCPK11으로 in vitro kinase assay를 실시한 결과, OsCPK11은 Ca²⁺-의존성 자가인산화 활성을 가짐을 보여 주었다. OsCPK11은 histone III-S를 인산화 하였으며, 카이네즈 활성의 최적 pH는 7.5-8.0이었다. Native OsCPK11은 이전에 연구된 재조합 OsCPK11과 몇 가지 생화학적 특징을 공유하였는데, 둘 다 Ca²⁺-의존성 자가인산화 활성을 나타냈다. 또한, 둘 모두 카이네즈 활성을 위한 기질로서 histone III-S를 선호하였으며, Ca²⁺ 의존성을 보여 주었다.