• Parasites, Hosts and Diseases
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• 제54권6호
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• pp.759-768
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• 2016

Cystic echinococcosis (CE) treatment urgently requires a novel drug. The p38 mitogen-activated protein kinases (MAPKs) are a family of Ser/Thr protein kinases, but still have to be characterized in Echinococcus granulosus. We identified a 1,107 bp cDNA encoding a 368 amino acid MAPK protein (Egp38) in E. granulosus. Egp38 exhibits 2 distinguishing features of p38-like kinases: a highly conserved T-X-Y motif and an activation loop segment. Structural homology modeling indicated a conserved structure among Egp38, EmMPK2, and H. sapiens $p38{\alpha}$, implying a common binding mechanism for the ligand domain and downstream signal transduction processing similar to that described for $p38{\alpha}$. Egp38 and its phosphorylated form are expressed in the E. granulosus larval stages vesicle and protoscolices during intermediate host infection of an intermediate host. Treatment of in vitro cultivated protoscolices with the p38-MAPK inhibitor ML3403 effectively suppressed Egp38 activity and led to significant protoscolices death within 5 days. Treatment of in vitro-cultivated protoscolices with $TGF-{\beta}1$ effectively induced Egp38 phosphorylation. In summary, the MAPK, Egp38, was identified in E. granulosus, as an anti-CE drug target and participates in the interplay between the host and E. granulosus via human $TGF-{\beta}1$.

• Preventive Nutrition and Food Science
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• 제19권3호
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• pp.136-144
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• 2014

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This study was conducted to evaluate the effects of Platycarya strobilacea S. et Z. (PSE) extract on mouse hair growth and to determine the mechanism of action of PSE. PSE was purchased and its antioxidant activities, such as electron donating ability, total polyphenol content, and flavonoid content were tested. Toxicity during topical treatment was determined by the CCK-8 assay, a cell viability test. Fifteen 4-week-old male C57BL/6 mice were assigned to receive one of three treatments: dimethyl sulfoxide (negative control), minoxidil (positive control) or PSE. Test materials were topically applied to the shaved dorsal skin of each mouse daily for 3 weeks. After 21 days, we observed skin tissue hair follicle morphology and length, mast cell number, and stem cell factor (SCF) expression using hematoxylin and eosin (H&E), toluidine blue, and immunohistochemical staining, respectively. Furthermore, the expression of cytokines involved in hair growth [i.e., insulin-like growth factor (IGF)-1, keratinocyte growth factor (KGF), and transforming growth factor (TGF)-${\beta}1$] was determined by PCR. PSE was found to have very high antioxidant activity. The cell viability rate of PSE-treated mice was markedly higher than that of mice in the control group. We also observed an increase in hair follicle length, strong SCF staining, and a decrease in mast cell number in the PSE group. In addition, PSE-treated mice had higher IGF-1 and KGF expression and lower TGF-${\beta}1$ expression than mice in the minoxidil-treated group. These results suggest that topical application of PSE promotes hair growth by intensifying SCF, suppressing mast cell production, and increasing hair growth-promoting cytokine expression.

• 대한한의학회지
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• 제31권1호
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• pp.138-152
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• 2010

Objective: Yikgeebohyul-tang (Yiqib$\check{u}$xu$\grave{e}$-t$\bar{a}$ng) is one of the representative prescriptions for invigoration of vitality and nourishing of the blood. This study was carried out to investigate the effects of Yikgeebohyul-tang (Yiqib$\check{u}$xu$\grave{e}$-t$\bar{a}$ng) on hair regrowth and cytokine changes in a shaving model of C57BL6 mice. Method: Five-week-old mice were acclimated for 1 week at a temperature between $21-23^{\circ}C{\acute{E}}$, 40-60% relative humidity, and 12h of a light/dark cycle before beginning of the experiment. There were two groups including normal saline (control) and a positive control of oral intake of Yikgeebohyul-tang (Yiqib$\check{u}$xu$\grave{e}$-t$\bar{a}$ng) extract (sample) in 18 female mice. The test compounds were topically treated once a day over 14 days. The hair regrowth was photographically and histologically determined during the experimental period of 14 days. Revelation of TGF-${\beta}1$ and EGF were also determined using immunohistochemistry. In addition to that, IFN-$\gamma$, IL-4 and IL-10 were determined in serum. Results: Hair regrowth in the sample group was promoted earlier and faster than the control group, as shown by concentrations of hairs and thick-hair ratio in the sample group. TGF-${\beta}1$ was not revealed in either control or sample group. EGF was strongly positive in out root sheath of some thick hair of the sample group. Serum IFN-$\gamma$ was significantly decreased in the sample group compared with the control group at 7 experimental days. However, it was not significantly decreased at 14 experimental days. Serum IL-4 was significantly increased in the sample group compared with the control group at 7 experimental days. However, it was not significantly decreased at 14 experimental days. Serum IL-10 was decreased in the sample group compared with the control group, but with no real statistical significance. Conclusion: These results suggest that Yikgeebohyul-tang (Yiqib$\check{u}$xu$\grave{e}$-t$\bar{a}$ng) has hair growth promoting activity and it can be used for treatment of alopecia. Also, these effects relate to EGF revelation of hair roots, a decrease in serum IFN-$\gamma$, and an increase of serum IL-4.

• Reproductive and Developmental Biology
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• 제32권3호
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• pp.167-173
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• 2008

Somatic cells such as oviduct epithelial cell, uterine epithelial cell, cumulus-granulosa cell and buffalo rat river cell has been used to establish an effective culture system for bovine embryos produced in in vitro. But nitric oxide (NO) metabolites secreted from somatic cells were largely arrested the development of bovine in vitro matured/ in vitro fertilized (IVM/IVF) embryos, suggesting that NO was induced the embryonic toxic substance into culture medium. The objective of this study was to investigate whether BOEC co-culture system can ameliorate the NO-mediated oxidative stress in the culture of bovine IVM/IVF embryos. Therefore, we evaluated the developmental rate of bovine IVM/IVF embryos under BOEC co-culture system in the presence or absence of sodium nitroprusside (SNP), as a NO donor, and also detected the expression of growth factor (TGF-$\beta$, EGF and IGFBP) and apoptosis (Caspase-3, Bax and Bcl-2) genes. The supplement of SNP over 5 uM was strongly inhibited blastocyst development of bovine IVM/IVF embryos than in control and 1 uM SNP group (Table 2). The developmental rates beyond morulae stages of bovine IVM/IVF embryos co-cultured with BOEC regardless of SNP supplement (40.4% in 5 uM SNP+ BOEC group and 65.1% in BOEC group) were significantly increased than those of control (35.0%) and SNP single treatment group (23.3%, p<0.05: Table 3). The transcripts of Bax and Caspase-3 genes were detected in all experiment groups (1:Isolated fresh cell (IFC), 2:Primary culture cell (PCC), 3:PCC after using the embryo culture, 4: PCC containing 5 uM SNP and 5: PCC containing 5 uM SNP after using the embryo culture), but Bcl-2 gene was not detected in IFC and PCC (Fig. 1). In the expression of growth factor genes, TGF-$\beta$ gene was found in all experimental groups, and EGF and IGFBP genes were not found in IFC and PCC (Fig. 2). These results indicate that BOEC co-culture system can increase the development beyond morula stages of bovine IVM/IVF embryos, possibly suggesting the alleviation of embryonic toxic substance like nitric oxide.

• 한국임상수의학회지
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• 제26권5호
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• pp.433-440
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• 2009

High levels of inflammatory cytokines were proposed contributors to the pathogenesis of a various inflammatory skin disorders. Therefore, investigating the immune response of the inflammatory skin disorder allows a better understanding of pathogenesis of a various inflammatory skin disorders and therapeutic approaches. The aim of this study was to analyze of the immune response in dogs with chronic inflammatory skin disease. To this aim, the present study evaluated relative mRNA expression of canine $IFN-{\gamma}$, IL-4, $TGF-{\beta}$ and IL-10 using TaqMan realtime PCR assays and semi-quantitative RT-PCR in freshly isolated peripheral blood mononuclear cells from twenty dogs with chronic inflammatory skin disease and ten normal dogs. The relative mRNA expression levels of IL-4 mRNA were significantly higher in dogs with chronic inflammatory skin disease than those in normal dogs (P < 0.01). The results of present study also showed a tendency towards increased expression of IL-10 transcripts in dogs with chronic inflammatory skin disease. However, there were no significant differences in the levels $IFN-{\gamma},\;TGF-{\beta}$ between normal and chronically inflammed dogs. In addition, the concentration of serum IgE was significantly increased in dogs with chronic inflammatory skin disease compared with those in normal dogs (P < 0.01). In histopathological examination, we found that there were markedly increased mast cell counts in chronically inflammed dogs (P < 0.05). These results suggest that the pathogenesis of chronic inflammatory skin disease might be associated with a T-cell mediated inflammatory responses characterized by a Th2-skewed immune response. Based on these results, the modulation of Th1/Th2 balance may be an effective therapeutic strategy for the treatment of chronic inflammatory skin disease.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제23권4호
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• pp.346-355
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• 2020

Purpose: Peroxisome proliferator-activated receptor gamma (PPAR-γ) has a key role in hepatic fibrogenesis by virtue of its effect on the hepatic stellate cells (HSCs). Although many studies have shown that PPAR-γ agonists inhibit liver fibrosis, the mechanism remains largely unclear, especially regarding the cross-talk between PPAR-γ and other potent fibrogenic factors. Methods: This experimental study involved 25 male Wistar rats. Twenty rats were subjected to bile duct ligation (BDL) to induce liver fibrosis, further divided into an untreated group (BDL; n=10) and a group treated with the PPAR-γ agonist thiazolidinedione (TZD), at 14 days post-operation (BDL+TZD; n=10). The remaining 5 rats had a sham operation (sham; n=5). The effect of PPAR-γ agonist on liver fibrosis was evaluated by histopathology, protein immunohistochemistry, and mRNA expression quantitative polymerase chain reaction. Results: Histology and immunostaining showed markedly reduced collagen deposition, bile duct proliferation, and HSCs in the BDL+TZD group compared to those in the BDL group (p<0.001). Similarly, significantly lower mRNA expression of collagen α-1(I), matrix metalloproteinase-2, platelet-derived growth factor (PDGF)-B chain, and connective tissue growth factor (CTGF) were evident in the BDL+TZD group compared to those in the BDL group (p=0.0002, p<0.035, p<0.0001, and p=0.0123 respectively). Moreover, expression of the transforming growth factor beta1 (TGF-β1) was also downregulated in the BDL+TZD group (p=0.0087). Conclusion: The PPAR-γ agonist inhibits HSC activation in vivo and attenuates liver fibrosis through several fibrogenic pathways. Potent fibrogenic factors such as PDGF, CTGF, and TGF-β1 were downregulated by the PPAR-γ agonist. Targeting PPAR-γ activity may be a potential strategy to control liver fibrosis.

• Parasites, Hosts and Diseases
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• 제47권2호
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• pp.145-151
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• 2009

This study examined the association of cytokine gene polymorph isms with intrahepatic bile duct wall fibrosis in human clonorchiasis. A total of 240 residents in Heilongjiang, China underwent ultrasonography, blood sampling, and stool examination. Single nucleotide polymorphism (SNP) sites for $IFN-{\gamma}$ (+874 T/A), IL-10 (-1,082 G/A, -819 C/T, -592 C/A), $TNF-{\alpha}$ (-308 G/A), and $TGF-{\beta}1$ (codon 10 T/C, codon 25 G/C) genes were observed with the TaqMan allelic discrimination assay. No significant correlation was observed between individual cytokine gene polymorphisms and intrahepatic duct dilatation (IHDD). Among individuals with clonorchiasis of moderate intensity, the incidence of IHDD was high in those with $IFN-{\gamma}$ intermediate-producing genotype, +874AT (80.0%, P=0.177), and in those with $TNF-{\alpha}$ low-producing genotype, -308GG (63.0%, P=0.148). According to the combination of $IFN-{\gamma}$ and $TNF-{\alpha}$ genotypes, the risks for IHDD could be stratified into high (intermediate-producing $IFN-{\gamma}$ and low producing $TNF-{\alpha}$), moderate, and low (low-producing $IFN-{\gamma}$ and high producing $TNF-{\alpha}$) risk groups. The incidence of IHDD was significantly different among these groups (P=0.022): 88.9% (odds ratio, OR=24.0) in high, 56.5% (OR=3.9) in moderate, and 25.0% (OR=1) in low risk groups. SNP of $IFN-{\gamma}$ and $TNF-{\alpha}$ genes may contribute to the modulation of fibrosis in the intrahepatic bile duct wall in clonorchiasis patients.

• 대한임상검사과학회지
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• 제47권3호
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• pp.117-124
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• 2015

중간엽줄기세포는 손상된 관절연골 치유능력을 가지고 있어 줄기세포 치료 분야에서 대표적인 성체줄기세포로 알려져 있다. 자외선이 조사된 생체 친화성 필름 조성물인 DTOPV (S-triazine bridged p-phenylenevinylene)는 친수성 특성의 표면을 가진 형광 화합물이다. 이전의연구에서 물질표면의 습윤성과 친수성이 세포부착 및 증식에 중요한 역할을 하는 것이 확인 되었으며, 이번 연구에서는 DTOPV를 이용하여 중간엽줄기세포의 연골분화능을 향상시키고자 하였다. 일반 배양용기로 사용하고 있는 TCPS (tissue culture polystyrene)와 자외선이 조사된 패턴된 DTOPV 필름을 이 실험에 사용하였고 TGF (transforming growth factor)-${\beta}3$가 포함된연골분화배지로 중간엽줄기세포를 2주동안 분화유도를 하였다. TCPS에서 배양된 중간엽줄기세포는 단층으로 자라면서 분화가 유도된 반면, 자외선이 조사된 DTOPV 필름 위에서 배양된 세포는덩어리진 구형으로 형태가 변하였으며, 연골세포에 특이적으로 염색되는 Safranine O 염색으로 DTOPV 조건에서 더 붉게 염색됨을 관찰하였다. 또한 연골세포 특이적인 유전자인 Type II collage이 DTOPV 조건에서 더 강하게 발현되는 것을 확인함으로써 TCPS보다 DTOPV에서 연골세포로 분화가 향상된 것을 알 수 있었다. 따라서 자외선이 조사된 생체 친화성 필름 조성물인 DTOPV을 이용한 경우에 일반 배양용기보다 빠르게 연골분화가 이루어짐을 알 수 있었다. 결론적으로 향후 조직공학 분야에서 DTOPV가 중간엽줄기세포의 효과적인 연골분화 물질로서 활용될 수 있는 가능성을 확인 하였으며, 더 나아가 약물 스크리닝과 같은 진단분야에 활용될 수 있음을 알 수 있었다.

• 생명과학회지
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• 제24권1호
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• pp.61-66
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• 2014

본 연구는 면역억제 마우스에서 양강 에탄올 추출물이 면역증강에 미치는 영향을 평가해보았다. 마우스에 cyclophosphamide를 2회 복강 주사한 후 양강 추출물을 30, 100, 300 mg/kg 용량으로 4주간 경구투여 한 후, 체중 및 면역장기 무게, 비장세포수, 혈청 사이토카인 농도 및 혈청 면역글로불린의 농도를 측정하였다. 실험 결과 체중과 비장세포수는 양강 추출물 투여 시 대조군과 비교하여 유의한 차이를 보이지 않았다. 혈청의 IL-2, TGF-${\beta}$ 및 IFN-${\gamma}$ 농도는 AO 100군에서 대조군에 비해 유의적인 증가를 보였고(p<0.05) IL-4 농도는 실험군에서 유의한 차이를 보이지 않았다. 혈청 내 IgM의 농도는 대조군에 비해 양강 추출물 투여군 모두에서 유의적으로 증가하였고 (p<0.05), IgA의 농도는 양강 추출물 투여군에서 증가하는 경향을 보였는데 특히 AO100군에서 유의성 있게 증가하였다(p<0.05). 본 연구 결과는 양강 에탄올 추출물은 혈청 내 사이토카인 농도와 면역글로불린 농도를 증가시켜 면역력 증강에 기여할 것으로 보이며 특히 100 mg/kg을 투여하였을 때 효과가 가장 큰 것으로 나타났다.

• Journal of Periodontal and Implant Science
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• 제31권3호
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• pp.513-529
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• 2001

For reconstruction of the bony defect, various artificial substitutes were developed. Among them, there has been a study of calcium phosphate coated bone substitutes for increasing attachment of osteoblasts in vivo. The purpose of this study was to evaluate the effects of serum and platelet-rich plasma (PRP) on calcium phosphate coated culture plate for the initial attachment, proliferation and activity of osteoblasts. After sampling the blood from white rats and concentrating by centrifugation, the amount of attachment of PDGF-BB and $TGF-{\beta}$ on the calcium phosphate coated culture plate was measured. Cultured HOS and ROS 17/2.8 cell was measured on attachment level and proliferation rate of osteoblasts. Alkaline phosphatase activity of HOS and ROS 17/2.8 cell was measured for studying on the activating rate of osteoblast. 1. Counting the amount of platelets of seperated plasma and PRP, the average number of platelets was 177,003 $cell/{\mu}l$ in plasma, and 1,656,062 $cell/{\mu}l$ in PRP, which was about 9 times as high as in plasma. 2. Amount of PDGF-BB deposited at calcium phosphate coated plate had increased by the total amount of plasma and PRP on the culture plate, whereas $TGF-{\beta}$had been deposited on the plate only when treated by $50{\mu}{\ell}$ of PRP(p<0.01). 3. After plating serum and PRP for 3 hours, we attached with HOS and ROS17/2.8 cell for 1 hour and 4 hours. There were no significant difference of the attachment between serum and control group, whereas there were significantly difference of the attachment between depositioning of PRP and control group. 4. After attaching plasma and PRP for 3 hours, cell number has much increased when HOS and ROS17/2.8 cell had been cultured for 48 hours(p<0.05). 5. After attaching plasma and PRP for 3 hours, concentration of alkaline-phosphatase has increased when HOS and ROS17/2.8 cell had been cultured for 48 hours(p<0.01). These results suggested that PRP affected on initial cell attachment rather than proliferation and activation of osteoblasts at calcium phosphate coated plate.