• 대한한의학방제학회지
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• 제30권2호
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• pp.45-57
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• 2022

Objectives : Yukil-san (YIS, 六一散; Liu yi san) is composed of Talcum and Glycyrrhizae Radix, the name is said to be derived from the proportion of the two herbal components of the formula. The YIS originated from 'Formulas from the discussion illuminating the Yellow Emperor's Basic Question'(黃帝素問宣明論方; Huang di su wen xuan ming lun fang) written by Liu Wan-Su (劉完素). YIS could clear summerheat, resolve dampness, and augment the qi. This formula may be used to treat the common cold, influenza, acute gastroenteritis, cystitis, urethritis and bacillary dysentery. But, there is insufficient of study about the effects of YIS on the anti-inflammatory activities. The present study evaluated the anti-inflammatory effects of YIS on lipopolysaccharide (LPS)-activated RAW 264.7 cells. Methods : Cell viability was assessed by MTT assay and nitric oxide (NO) was evaluated by measuring the nitrite content in culture medium. Pro-inflammatory cytokines such as tumor necrosis factor-α, interleukin-1β and IL-6 were quantified by ELISA kit. The expression of proteins related with nuclear factor-κB (NF-κB) pathway and inducible NO synthase (iNOS) were assessed by western blot analysis. Results : YIS significantly inhibited the expression of iNOS increased by LPS, and thus significantly inhibited the production of NO. In addition, YIS significantly inhibited pro-inflammatory cytokines. In the regulation of inflammation, NF-κB pathway plays a crucial role. YIS inhibited the expression of p-IκBα and thus inhibited the translocation of NF-κB to the nucleus. Conclusions : These results suggest that YIS ameliorates inflammatory response in LPS-activated RAW 264.7 cells through the inhibition of inflammatory mediators, via suppression of the NF-κB pathway. Therefore, this study provides objective evidence for the anti-inflammatory effect of YIS including the underlying mechanisms.

• 대한본초학회지
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• 제32권5호
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• pp.1-6
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• 2017

Objectives : Corni Fructus (CF) is traditional herbal medicine used on polyuria, low back pain, and tinnitus. This study aimed to evaluate inhibits skin wrinkle formation effect of CF. Methods : To evaluate the produce inhibition effect of CF, SD-rats were distributed into four groups; normal rats (Nor), AGEs (advenced glycation end product)-induced rats (Con), AGEs-induced rats treated with 100mg/kg CF (CF). To induce AGEs, streptozotocin (50mg/kg) was administered intraperitoneally and after 3 days oral administrated 100mM methyl glyoxal for 3 weeks. Results : The oral administration of CF suppressed the reactive oxygen specis (ROS) in serum. The AGEs in skin tissues was significantly reduced through treatment of CF. Furthermore, the expressions of AGEs related proteins such as polyclonal anti-$N^e$-(carboxymethyl)lysine (CML), anti-$N^e$-(carboxyethyl)lysine (CEL), AGE receptors (RAGE) were decreased in CF treated group compared with the control group in skin tissues. Inflammation-related proteins such as Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB), monocyte chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6) reduced in CF treatment group than control group. AGE-induced rats exhibited that the significant decreased collagen however, CF treatment (100mg/kg of body weight) up regulated collagen by improved the expression levels of skin fibril-related genes such as Matrix metalloproteinase (MMP-1). Conclusion : Taken together, our study suggests that CF regulates ROS to prevent accumulation of AGEs and inhibits skin wrinkles. Our finding indicate that CF may be an effective agent for inhibits AGEs formation, and improved skin wrinkle.

• 동의생리병리학회지
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• 제36권3호
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• pp.89-93
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• 2022

Earthing, caused by direct skin contact with the Earth's surface, is used to reduce the symptoms of inflammation (fever, fever, swelling and pain). However, there is little evidence to support the anti-inflammatory effects of earthing mattresses. Therefore, this study was conducted to investigate whether anti-inflammatory effect of earthing mattress using an in vivo animal model. The anti - inflammatory effect was evaluated by measuring ear thickness and foot volume in 12-O-tetradecanoylphorbol-13 acetate (TPA) - induced ear edema and carrageenan - induced paw edema model, respectively. Balb/c mouse in carrageenan paw edema model showed significant anti - inflammatory effect in the group treated with earthing mattress for 4 hours or 24 hours for 3 days. For females, the anti-inflammatory effect was greater when the earthing mattress was added to the mattress than the mattress alone treatment. From the above results, it was found that the female responds more to the effect of the earthing as well as the mattress effect. In addition, when the male and female Balb/c mice were exposed to mattresses and earthing mattresses for 24 h for 3 days, respectively, the mattress and earthing mattresses showed significant inhibition of IL (Interleukin)-1β levels compared to the control. In the TPA ear edema model, Balb/c mouse showed significant anti - inflammatory effect in the group treated with the earthing mattress for 4 hours or 24 hours for 3 days. Both males and females showed more anti-inflammatory effects when they were exposed to earthing mattresses with mattresses added to the mattresses. From the above results, it was found that both male and female respond to the effect of earthing as well as the mattress effect in the TPA ear edema model. In conclusion, in this study, we have verified that earthing mattress shows inhibitory effects on TPA and carrageenan-induced inflammation. From these results, it is suggested that the anti-inflammatory effect can be expected by applying the earthing mattress to patients suffering from inflammatory diseases. However, there is a need to pinpoint exactly how the earthing mattress relieves inflammation, and further research is needed to investigate the mechanism.

• Animal Bioscience
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• 제36권6호
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• pp.851-860
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• 2023

Objective: This study aims to evaluate the target genes of gga-miR-20a-5p and the regulated immune responses in the chicken macrophage cell line, HD11, by the exosome-mediated delivery of miR-20a-5p. Methods: Exosomes were purified from the chicken macrophage cell line HD11. Then, mimic gga-miR-20p or negative control miRNA were internalized into HD11 exosomes. HD11 cells were transfected with gga-miR-20a-5p or negative control miRNA containing exosomes. After 44 h of transfection, cells were incubated with or without 5 ㎍/mL poly(I:C) for 4 h. Then, expression of target genes and cytokines was evaluated by quantitative realtime polymerase chain reaction. Results: Using a luciferase reporter assay, we identified that gga-miR-20a-5p directly targeted interferon gamma receptor 2 (IFNGR2), mitogen-activated protein kinase 1 (MAPK1), mitogen-activated protein kinase kinase kinase 5 (MAP3K5), and mitogen-activated protein kinase kinase kinase 14 (MAP3K14). Moreover, the exosome-mediated delivery of gga-miR-20a-5p successfully repressed the expression of IFNGR2, MAPK1, MAP3K5, and MAP3K14 in HD11 cells. The expressions of interferon-stimulated genes (MX dynamin like GTPase 1 [MX1], eukaryotic translation initiation factor 2A [EIF2A], and oligoadenylate synthase-like [OASL]) and proinflammatory cytokines (interferon-gamma [IFNG], interleukin-1 beta [IL1B], and tumor necrosis factor-alpha [TNFA]) were also downregulated by exosomal miR-20a-5p. In addition, the proliferation of HD11 cells was increased by exosomal miR-20a-5p. Conclusion: The exosome-mediated delivery of gga-miR-20a-5p regulated immune responses by controlling the MAPK and apoptotic signaling pathways. Furthermore, we expected that exosomal miR-20a-5p could maintain immune homeostasis against highly pathogenic avian influenza virus H5N1 infection by regulating the expression of proinflammatory cytokines and cell death.

• Journal of Veterinary Science
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• 제23권1호
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• pp.2.1-2.18
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• 2022

Background: Co-infections of the porcine reproductive and respiratory syndrome virus (PRRSV) and the Haemophilus parasuis (HPS) are severe in Chinese pigs, but the immune response genes against co-infected with 2 pathogens in the lungs have not been reported. Objectives: To understand the effect of PRRSV and/or HPS infection on the genes expression associated with lung immune function. Methods: The expression of the immune-related genes was analyzed using RNA-sequencing and bioinformatics. Differentially expressed genes (DEGs) were detected and identified by quantitative real-time polymerase chain reaction (qRT-PCR), immunohistochemistry (IHC) and western blotting assays. Results: All experimental pigs showed clinical symptoms and lung lesions. RNA-seq analysis showed that 922 DEGs in co-challenged pigs were more than in the HPS group (709 DEGs) and the PRRSV group (676 DEGs). Eleven DEGs validated by qRT-PCR were consistent with the RNA sequencing results. Eleven common Kyoto Encyclopedia of Genes and Genomes pathways related to infection and immune were found in single-infected and co-challenged pigs, including autophagy, cytokine-cytokine receptor interaction, and antigen processing and presentation, involving different DEGs. A model of immune response to infection with PRRSV and HPS was predicted among the DEGs in the co-challenged pigs. Dual oxidase 1 (DUOX1) and interleukin-21 (IL21) were detected by IHC and western blot and showed significant differences between the co-challenged pigs and the controls. Conclusions: These findings elucidated the transcriptome changes in the lungs after PRRSV and/or HPS infections, providing ideas for further study to inhibit ROS production and promote pulmonary fibrosis caused by co-challenging with PRRSV and HPS.

• 농업생명과학연구
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• 제50권2호
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• pp.139-150
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• 2016

말뼈추출물은 다양한 골질환의 예방과 치료에 탁월한 효능이 있다고 이전에 보고되었다. 하지만 말뼈추출물의 다른 약리학적 효능에 대해서는 아직 자세히 밝혀지지 않고있다. 본 연구에서는 말뼈추출물이 중요한 항산화 인자인 hemeoxygenase-1(HO-1)의 발현을 상승시킬 수 있는지, 만약 발현이 증가한다면 HO-1의 상향 조절이 대식세포에서 항염증 효과를 매개할 수 있는지에 관하여 조사하였다. 이를 위해서 nitric oxide(NO) 농도측정, 세포 생존능 측정, DPPH 라디칼 소거능 검사를 시행하였다. 또한 염증성 사이토카인 유전자 발현과 단백질 발현을 측정하기 위해 real time PCR과 Western blotting을 시행하였다. 말뼈추출물은 lipopolysaccharide(LPS, 0.1㎍/ml)로 자극한 대식세포주인 RAW264.7 세포에서 어떠한 세포독성 없이 NO의 생성을 유의성 있게 억제하였으며 inducible nitric oxide(iNOS)와 cyclooxygenase 2(COX-2)의 발현을 억제하였다. 뿐만 아니라 말뼈 추출물은 염증성 사이토카인인 tumor necrosis factor(TNF)-α와 interleukin(IL)-1β의 발현을 억제하였으며 ERK, JNK 및 p-38 MAPK의 단백질 인산화를 억제하였다. 그리고 말뼈추출물은 HO-1과 NF-E2-related factor-2(Nrf-2) 의 발현을 증가시켰고 이것은 말뼈추출물이 가지고 있는 항 염증효과를 매개할 수 있는 것으로 보인다. 즉, 말뼈추출물이 HO-1의 발현을 상향 조절한 반면 ERK1/2의 신호전달 경로에 손상을 주는 것으로 확인되었으며 이러한 말뼈추출물의 효과가 최종적으로 세포손상과 세포의 과산화 자극으로부터 세포를 보호 할 수 있는 것으로 사료된다.

• 대한본초학회지
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• 제38권4호
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• pp.21-29
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• 2023

Background : The purpose of this study was to investigate the anti-inflammatory properties of stems and leaves of Eleutherococcus senticosus (Rupr. & Maxim.) Maxim. (ES) from Gangwon-do. Methods and Results : Stems and leaves of ES were collected from two areas in Gangwon-do: Cheorwon-gun and Samcheok-si. Samples were extracted with water by using the pressurized liquid extraction method and with 70% prethanol A by using the heat reflux extraction method. The anti-inflammatory effects of ES were evaluated through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide(MTT), lactate dehydrogenase(LDH) assay, nitric oxide(NO) assay, enzyme-linked immunosorbent assay(ELISA), and Western blot analysis in RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS). 1) Results showed that ES leaf extractions were not cytotoxic at a concentration of up to 30 ㎍/㎖. The leaves of 70% prethanol A extractions of ES(30 ㎍/㎖) inhibited NO, interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) production and decreased the protein level of cyclooxygenase 2(COX-2). There was no significant change in the protein level of inducible nitric oxide synthase(iNOS). The stem extractions of ES did not exhibit anti-inflammatory effects. Conclusions : In this study, the leaves of 70% prethanol A extractions of ES demonstrated anti-inflammatory effect on RAW 264.7 macrophages. The 70% prethanol A extractions have a relatively higher anti-inflammatory effect on RAW 264.7 macrophages than water extractions.

• 대한한방내과학회지
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• 제44권3호
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• pp.402-417
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• 2023

Objective: Liver fibrosis is a highly conserved wound-healing response and the final common pathway of chronic inflammatory injury. This study aimed to evaluate the potential anti-fibrotic effect of the combination of Rhei Radix et Rhizoma water extract (RW) and silymarin in a thioacetamide (TAA)-induced liver fibrosis model. Methods: The liver fibrosis mouse model was established through the intraperitoneal injection of TAA (1 week 100 mg/kg, 2-3 weeks 200 mg/kg, 4-8 weeks 400 mg/kg) three times per week for eight weeks. Animal experiments were conducted in five groups; Normal, Control (TAA-induced liver fibrosis mice), Sily (silymarin 50 mg/kg), RSL (RW 50 mg/kg+silymarin 50 mg/kg), and RSH (RW 100 mg/kg+silymarin 50 mg/kg). Biochemical analyses were measured in serum, including aspartate aminotransferase (AST), alanine aminotransferase (ALT), malondialdehyde (MDA), and ammonia levels. Liver inflammatory cytokines and fibrous biomarkers were measured by Western blot analysis, and liver histopathology was evaluated through tissue staining. Results: A significant decrease in the liver function markers AST and ALT and a reduction in ammonia and total bilirubin were observed in the group treated with RSL and RSH. Measurement of reactive oxygen species and MDA revealed a significant decrease in the RSL and RSH administration group compared to the TAA induction group. The expression of extracellular matrix-related proteins, such as transforming growth factor β1, α-smooth muscle actin, and collagen type I alpha 1, was likewise significantly decreased. All drug-administered groups had increased matrix metalloproteinase-9 but a decreasing tissue inhibitor of matrix metalloproteinase-1. RSL and RSH exerted a significant upregulation of NADPH oxidase 2, p22^phox, and p47^phox, which are oxidative stress-related factors. Furthermore, pro-inflammatory proteins such as cyclooxygenase 2 and interleukin-1β were markedly suppressed through the inhibition of nuclear factor kappa B activation. Conclusions: The administration of RW and silymarin suppressed the NADPH oxidase factor protein level and showed a tendency to reduce inflammation-related enzymes. These results suggest that the combined administration of RW and silymarin improves acute liver injury induced by TAA.

• Journal of Nutrition and Health
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• 제57권1호
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• pp.88-104
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• 2024

본 연구는 국민건강영양조사 2015년부터 2018년도 자료를 활용하여 우리나라 만 19세 이상 성인의 KHEI와 복부비만의 관련성 및 hs-CRP의 매개 효과를 분석한 연구이다. 한국 성인 남성의 경우 허리둘레 90 cm 이상, 여성은 허리둘레 85 cm 이상인 경우에 복부비만군으로 정의하였다. 남성에서 복부비만군은 2,587명, 정상군은 5,007명이였고 여성의 복부비만군은 2,869명, 정상군은 7,307명이였다. KHEI, hs-CRP, 복부비만의 연관성 분석에서는 모든 공변량을 보정하였을 때 남성의 KHEI는 복부비만과 유의한 연관이 없었지만, hs-CRP와 음의 연관성이 있었다. 여성의 경우, 복부비만과 hs-CRP 모두 음의 연관성이 관찰되었다. 또한, 남성과 여성에서 hs-CRP 위험군 (> 1.0 mg/L) 이 hs-CRP 정상군 (≤ 1.0 mg/L)보다 복부비만 유병률이 유의하게 높았다. Bootstrapping을 이용한 매개효과 분석에서는 KHEI와 복부비만 관계에서 hs-CRP가 매개 효과를 가지는 것으로 나타났으며 공변량을 단계적으로 보정하였을때 증가하는 양상이 나타났다. 결론적으로, 본 연구 결과는 균형잡힌 식생활를 통한 높은 식사의 질이 hs-CRP에 영향을 미쳐 복부비만 예방에 중요한 영향을 미칠 수 있음을 시사한다.

• Journal of Korean Neurosurgical Society
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• 제67권3호
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• pp.333-344
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• 2024

Objective : Markers of neuroinflammation during ischemic stroke are well characterized, but additional markers of neural damage are lacking. The study identified associations of behavioral disorders after stroke with histologic neural damage and molecular biological change. Methods : Eight-week-old, 25 g male mice of the C57BL/6J strain were subjected to middle cerebral artery occlusion (MCAO) to induce ischemic stroke. The control group was a healthy wild type (WT), and the experimental group were designed as a low severity MCAO1 and a high severity MCAO2 based on post-stroke neurological scoring. All groups underwent behavioral tests, realtime polymerase chain reaction, triphenyltetrazolium chloride (TTC) staining and Hematoxylin and Eosin staining. One-way analysis of variance was used to analyze statistical significance between groups. Results : In TTC staining, MCAO1 showed 29.02% and MCAO2 showed 38.94% infarct volume (p<0.0001). The pro-inflammatory cytokine interleukin (IL)-1β was most highly expressed in MCAO2 (WT 0.44 vs. MCAO1 2.69 vs. MCAO2 5.02, p<0.0001). From the distance to target in the Barnes maze test, WT had a distance of 178 cm, MCAO1 had a distance of 276 cm, and MCAO2 had a distance of 1051 (p=0.0015). The latency to target was 13.3 seconds for WT, 27.9 seconds for MCAO1, and 87.9 seconds for MCAO2 (p=0.0007). Prospero homeobox 1 (Prox1) was most highly expressed in MCAO2 (p=0.0004). Doublecortin (Dcx) was most highly expressed in MCAO2 (p<0.0001). Conclusion : The study demonstrated that histological damage to neural cells and changes in brain mRNA expression were associated with behavioral impairment after ischemic stroke. Prox1 and Dcx may be biomarkers of neural damage associated with long-term cognitive decline, and increased expression at the mRNA level was consistent with neural damage and long-term cognitive dysfunction.