• Journal of Nutrition and Health
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• 제53권1호
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• pp.1-12
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• 2020

본 연구에서는 고혈당으로 인해 유발되는 당뇨병성 신장병증의 대표적인 증상인 사구체 경화증을 완화시키는 nobiletin의 효능에 대해 알아보고자 하였다. 신장 세포인 HRMC를 이용하여 고혈당에서의 세포외 기질 축적 단백질의 발현과 경화에 관여하는 신호 전달 억제 효능을 확인한 결과 nobiletin은 고혈당의 자극에 의해 증가하는 섬유화 단백질인 collagen IV, fibronectin 그리고 CTGF의 발현을 억제하였으며, 여기에 관여하는 TGF-β1-Src-caveolin-1 신호 전달 경로를 통해 사구체 경화증을 억제하는 것을 확인하였다. 따라서 nobiletin은 고혈당으로 유도된 당뇨병성 신장병증에 있어 사구체 경화증을 예방하는 기능성 성분으로서의 활용 가능성을 확인하였다.

• Tuberculosis and Respiratory Diseases
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• 제82권2호
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• pp.133-142
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• 2019

Background: Idiopathic pulmonary fibrosis involves irreversible alveolar destruction. Although alveolar epithelial type II cells are key functional participants within the lung parenchyma, how epithelial cells are affected upon bleomycin (BLM) exposure remains unknown. In this study, we determined whether BLM could induce cell cycle arrest via regulation of Schlafen (SLFN) family genes, a group of cell cycle regulators known to mediate growth-inhibitory responses and apoptosis in alveolar epithelial type II cells. Methods: Mouse AE II cell line MLE-12 were exposed to $1-10{\mu}g/mL$ BLM and $0.01-100{\mu}M$ baicalein (Bai), a G1/G2 cell cycle inhibitor, for 24 hours. Cell viability and levels of pro-inflammatory cytokines were analyzed by MTT and enzyme-linked immunosorbent assay, respectively. Apoptosis-related gene expression was evaluated by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Cellular morphology was determined after DAPI and Hoechst 33258 staining. To verify cell cycle arrest, propidium iodide (PI) staining was performed for MLE-12 after exposure to BLM. Results: BLM decreased the proliferation of MLE-12 cells. However, it significantly increased expression levels of interleukin 6, tumor necrosis factor ${\alpha}$, and transforming growth factor ${\beta}1$. Based on Hoechst 33258 staining, BLM induced condensation of nuclear and fragmentation. Based on DAPI and PI staining, BLM significantly increased the size of nuclei and induced G2/M phase cell cycle arrest. Results of qRT-PCR analysis revealed that BLM increased mRNA levels of BAX but decreased those of Bcl2. In addition, BLM/Bai increased mRNA levels of p53, p21, SLFN1, 2, 4 of Schlafen family. Conclusion: BLM exposure affects pulmonary epithelial type II cells, resulting in decreased proliferation possibly through apoptotic and cell cycle arrest associated signaling.

• Tuberculosis and Respiratory Diseases
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• 제55권2호
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• pp.140-153
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• 2003

연구배경 : 결핵균 항원으로 세포 매개성 면역반응이 활성화되면 여러 종류의 cytokine이 분비되고 각기 다른 cytokine과 network system으로 작용하여 여러 병태생리적인 과정을 조절한다. 결핵 환자에서 염증반응, 조직파괴 및 질환의 중증도는 proinflammatory cytokine과 suppressive cytokine의 균형과 조합에 의하여 결정되며, 결핵 진행의 제한과 악화에 중요한 역할을 할 것으로 생각되고 있다. 따라서 결핵의 이환과정에서 cytokine의 분비 및 변화와 역할을 파악하는 것이 질환의 병태생리를 이해하는데 크게 도움이 될 수 있다. 대상 및 방법 : 활동성 폐결핵 83명, 기관지 결핵 10명의 치료전과 정상 대조군 20명에서 말초혈액을 채취하여 혈청을 분리하여 $-70^{\circ}C$에 보관하였고, sandwich ELISA 방법을 이용하여 혈청 IL-$1{\beta}$, IL-2, IL-4, IL-6, IL-10, IL-12(p40), TNF-${\alpha}$, IFN-${\gamma}$, TGF-${\beta}$를 측정하였다. 폐결핵 환자 83명을 ATS guideline에 따라 중증도를 분류하였고, 추적관찰 중 탈락자를 제외한 45명에서 초치료 2개월과 6개월 후 각각 혈청 sIL-$1{\beta}$, IL-2, IL-4, IL-6, IL-10, IL-12(p40), TNF-${\alpha}$, IFN-${\gamma}$, TGF-${\beta}$를 재측정하였다. 결 과 : 1) IL-$1{\beta}$, TNF-${\alpha}$, IFN-${\gamma}$는 폐결핵 환자에서 대조군에 비하여 증가된 경향을 보였고(p>0.05), IL-6는 폐결핵군에서 통계적으로 유의하게 증가되었다(p<0.05). TGF-${\beta}$는 폐결핵과 기관지 결핵환자에서의 분비가 대조군에 비하여 감소된 경향을 보였으며(p>0.05), IL-2, IL-12(p40), IL-4, IL-10은 대조군과 폐결핵 환자에서 별다른 차이를 보이지 않았다. 2) 기관지 결핵 환자에서 IL-6, TNF-${\alpha}$는 대조군에 비해 증가되고 TGF-${\beta}$는 감소된 경향을 보였다(p>0.05). 폐결핵에 비해 IL-12(p40)의 분비는 증가된 경향을 보였다. 3) 결핵의 중증도가 심할수록 IFN-${\gamma}$와 IL-6가 유의하게 증가하였고(P<0.05), 특히 중증군에서 현저하였다. 4) 폐결핵 환자에서 치료전 측정한 IL-$1{\beta}$, IL-6, TNF-${\alpha}$ 간 및 IL-2, IL-4, IL-12 간에는 강한 양의 상관관계를 보였다(p<0.01). 5) 폐결핵 환자 45명에서 측정한 IL-6와 IFN-${\gamma}$는 치료후 2개월과 6개월에 각각 통계적으로 유의하게 감소되었다(p<0.05). 결 론 : 이상의 결과로 결핵의 병태생리에 있어서 여러 cytokine간의 균형과 조합의 변화가 숙주의 염증, 조직파괴 및 결핵의 중증도와 관련되어 있을 것으로 생각되지만 결핵의 형태나 면역반응 정도에 따른 다양성을 보여서 결과의 해석과 cytokine 측정의 임상적 이용에 대한 연구가 더욱 필요할 것으로 사료된다.

• 생명과학회지
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• 제25권12호
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• pp.1425-1431
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• 2015

면역 시스템의 림프구는 B 림프구와 T 림프구 두 종류로 나눌 수 있다. B 림프구는 플라즈마 세포로 분화하여 항체를 생성하는 체액성 면역을 담당하며, T 림프구는 다른 세포나 세균을 죽이는 세포성 면역을 담당한다. 고전적으로 B 림프구와 T 림프구의 작용은 한 방향으로 이뤄졌다. T 림프구는 B 림프구의 분화를 촉진하고 면역글로불린종류의 전환을 조절한다. T 림프구가 부족한 경우 B 림프구의 부족을 초래함이 보고되어 있다. 그러나 최근에 역으로 B 림프구가 T 림프구의 분화와 활성을 조절할 수 있다는 보고가 있다. 예를 들어, B 림프구는 CD8+ T 림프구의 tolerance를 직접 조절할 수 있고, TGF-β의 분비를 통해 T 림프구의 anergy를 유도할 수 있다. 본 연구는 LPS에 의해 자극된 B 림프구가 수지상세포에서 IL-12의 분비를 억제하여 Th1 림프구의 분화를 억제할 수 있음을 보여준다. 이 억제는 B 림프구와 수지상세포의 직접적인 interaction에 의해 일어나는 것이 아니며 B 림프구가 수지상세포의 성숙을 조절하여 일어나는 것도 아니다. B 림프구에서 분비되는 soluble factor가 LPS에 의해 증가되는 수지상세포의 IL-12p35 transcription을 억제한다. 이 결과들은 B 림프구가 매개하는 새로운 면역억제 기전이 존재함을 보여준다. 이것은 고전적인 방향성을 가진 T 림프구에 의한 B 림프구 작용조절로 면역반응이 결정되는 것이 아니라 T 림프구와 B 림프구가 서로 작용을 하여 면역평형을 결정하는 기전이 존재함을 보여준다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권11호
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• pp.4437-4441
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• 2014

SMAD7 has been identified as a functional candidate gene for colorectal cancer (CRC). SMAD7 protein is a known antagonist of the transforming growth factor beta ($TGF-{\beta}$) signaling pathway which is involved in tumorigenesis. Polymorphisms in SMAD7 may thus alter cancer risk. The aim of this study was to investigate the influence of a SMAD7 gene polymorphism (rs2337107) on risk of CRC and clinicopathological features in an Iranian population. In total, 210 subjects including 105 patients with colorectal cancer and 105 healthy controls were recruited in our study. All samples were genotyped by TaqMan assay via an ABI 7500 Real Time PCR System (Applied Biosystems) with DNA from peripheral blood. The polymorphism was statistically analyzed to investigate the relationship with the risk of colorectal cancer and clinicopathological properties. Logistic regression analysis revealed that there was no significant association between rs2337107and the risk of colorectal cancer. In addition, no significant association between genotypes and clinicopathological features was observed (p value>0.05). Although there was not any association between genotypes and disorder, CT was the most common genotype in this population. This genotype prevalence was also higher in the patients with well grade (54.9%) and colon (72.0%) tumors. Our results provide the first evidence that this polymorphism is not a potential contributor to the risk of colorectal cancer and clinicopathological features in an Iranian population, and suggests the need of a large-scale case-control study to validate our results.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제38권4호
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• pp.204-211
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• 2012

Objectives: Dental implants installation in patients with diabetes remains controversial as altered bone healing around implants has been reported. And little is known about the biological factors involved in bone healing around implants. The present study aimed to investigate the biological markers around immediately placed implants in rats with controlled and uncontrolled diabetes. Materials and Methods: Twenty rats (40 sites) were divided into the control, insulin-treated and diabetic groups. The rats received streptozotocin (60 mg/kg) to induce diabetes; animals in the insulin-treated group also received three units of subcutaneous slow-release insulin. Two threaded titanium alloy implant ($1.2{\times}3mm$) were placed in the extraction socket of the both maxillary first molars and allowed for healing. Bone blocks including implant were harvested at 3 days, 1, 2 and 4 weeks. The levels of bone morphogenetic protein (BMP)-4, transforming growth factor (TGF)-${\beta}1$, osteocalcin (OC) and osteonectin (ON) were measured in the peri-implant osseous samples by RT-PCR. Results: The BMP-4 level increased immediately in all groups by day 3, then decreased abruptly in the control and the insulin-treated groups. However, by week 4, all groups showed mostly the same amount of BMP-4 expression. The level of TGF-${\beta}1$ also instantly increased by day 3 in the insulin-treated group. This level elevated again reaching the same values as the control group by week 4, but was not as high as the diabetic group. In addition, the expression of OC and ON in the control and insulin-treated groups was higher than that of the diabetic group at 2 weeks and 4 weeks, indicating active bone formation in these groups. Conclusion: The immediate placement of titanium implants in the maxilla of diabetic rat led to an unwanted bone healing response. Conclusively, the results of this study suggest that immediate implant insertion in patients with poorly controlled diabetes might be contraindicated.

• Journal of Periodontal and Implant Science
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• 제29권4호
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• pp.765-785
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• 1999

Bone morphogenetic protein-2/4 (BMP-2/4) are members of Transforming Growth $Factor-{\beta}\;(TGF-{\beta})$ superfamily and they may differentiate the osteoprogenitor cell and induce formation of cartilage and bone in vivo. This study was performed to investigate the effects of bone morphogenetic protein-2/4 on the characteristics of rat periodontal ligament cells(RPDL) and rat calvaria cells(RCV). In the control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 20% fetal bovine serum, $100{\mu}/ml$ penicillin, $100{\mu}/ml$ streptomycin. In the experimental groups, recombinant human bone morphogenetic protein-2/4 (25ng, 100ng, 250ng/ml) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 2, 3, 5, 7th day, the amount of total protein synthesis and alkaline phosphatase activity at 2, 5, 7th day. And also, the calcified nodule was examed. The results were as follows ; 1 . Both RCV and RPDL cells in both control and experimental groups proliferated during the entire experimental period, but there is no stastically significant difference according to the BMP-2/4 concentration. 2 . Amount of total protein synthesis of both cells in both groups was steadily increased until 5th day, but all experimental groups were significantly different from the control group at 7th day. 3. Alkaline phosphatase activity of both cells in both groups was increased during the entire experiment period. In RCV cells, the experimental group treated with 100ng/ml and 250ng/ml BMP-2/4 were significantly different from the control group at 7th day. In RPDL cells, the experimental group treated with 100ng/ml and 250ng/ml BMP-2/4 were significantly different from the control group at 5th day, and all experimental groups were significantly different from the control group at 7th day. 4. In the both of the cultured Rat Periodontal ligament and calvaria cell treated with BMP-2/4 to compared with control group, it revealed more rapid cell polarization, cell aggregation and hyperchromatic stained on HE agent, and even though only 1 day treated with BMP-2/4 both RPDL and RCV showed more rapid cell reaction than control group. More sensivitve cell reaction of RCV were observed than RPDL in this experiment. From the above results, we could conclude that BMP-2/4 influenced the induction, proliferation and differentiation of bone forming cells

• 대한한의학회지
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• 제30권4호
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• pp.13-27
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• 2009

Objectives: Trans-cinnamaldehyde (TCA) is the main component of Cinnamomi Ramulus and it has been reported that TCA inhibits inflammatory responses in various cell types. Inflammation-mediated neurological disorders induce the activation of macrophages such as microglia in brain, and these activated macrophages release various inflammation-related molecules, which can be neurotoxic if overproduced. In this study, we evaluated gene expression profiles using gene chip microarrays in lipopolysaccharide (LPS)-stimulated BV-2 cells to investigate the antiinflammatory effect of TCA on inflammatory responses in brain microglia. Methods: A negative control group was cultured in normal medium and a positive control group was stimulated with $1{\mu}g/ml$ in the absence of TCA. TCA group was pretreated with $10{\mu}g/ml$ before $1{\mu}g/ml$ LPS stimulation. The oligonucleotide microarray analysis was performed to obtain the expression profiles of 28,853 genes using gene chip mouse gene 1.0 ST array in this study. Results: In positive control group, 1522 probe sets were up-regulated in the condition of the cutoff value of 1.5-fold change and 341 genes with Unigene ID were retrieved. In TCA group, 590 probe sets were down-regulated from among 1522 probe sets and 33 genes with Unigene ID were retrieved, which included 6 inflammation-related genes. We found out that Id3 gene is associated with transforming growth factor-${\beta}$ (TGF-${\beta}$) signaling pathway and Klra8 gene is related to natural killer cell-mediated cytotoxicity pathway. Conclusions: The results mean that TCA inhibits inflammatory responses through down-regulating the expressions of inflammation-related genes in LPS-stimulated BV-2 cells.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제19권3호
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• pp.265-286
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• 1997

Bone morphogenetic proteins(BMPs) are a group of transforming growth factor beta(TGF-${\beta}$)-related factors and multifunctional proteins, especially the only known biologic factors capable of inducing endochondral bone formation at an extraskeletal site. This study was performed to investigate the effect of the partially purified porcine BMP(pBMP) at an ectopic site. PBMP was partially purified from porcine bone matrix and its activity was monitored by an in vivo bioassay. The purification method utilized extraction of the bone-inducing activity with 4M guanidine, followed by chromatography on heparin-Sepharose. Active fractions were assayed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. And the fractions were reconstituted with inactive insoluble collagenous bone matrix from rats, acid soluble type I collagen from rat tail and chondroitin-6-sulfate sodium salt and implanted into the pectroralis muscle pouches of Sprague-Dawley rats. And the carrier complex was implanted on the opposite side as control. The rats were sacrificed at the day of 1st, 3rd, 5th, 7th, 11th, 14th and 21st after implantation and examined histologically, radiologically and biochemically. And alkaline phosphatase activity and calcium content were used as indices of bone formation. The results were as follows ; 1. Active fractions were localized in a zone between 31 and 40 KDa on SDS-PAGE. 2. The implanted 3.0mg of the partially purified pBMP induced cartilage and bone in the muscle tissue of rats through an endochondral ossification process. 3. Inactive insoluble bone matrix, type I collagen and chondroitin-6-sulfate have functioned as carriers for pBMP, but revealed some foreign body reactions. 4. Soft X-ray didn't reveal significant change between the experimental and the control group. 5. The alkaline phosphatase activities in the experimental group of 5th, 7th, 11th, 14th and 21st were increased significantly compared with control (p<0.01) with the peak in the group of 11th day. 6. With time, the calcium content of the experimental group increased. And the calcium contents in the experimental group of 11th, 14th and 21st were increased significantly compared with control (p<0.01).

• 대한약침학회지
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• 제18권1호
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• pp.72-78
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• 2015

Objectives: The stomach is a sensitive digestive organ that is susceptible to exogenous pathogens from the diet. In response to such pathogens, the stomach induces oxidative stress, which might be related to the development of both gastric organic disorders such as gastritis, gastric ulcers, and gastric cancer, and functional disorders such as functional dyspepsia. This study was accomplished to investigate the effect of Ganoderma lucidum pharmacopuncture (GLP) on chronic gastric ulcers in rats. Methods: The rats were divided into 4 groups of 8 animals each: the normal, the control, the normal saline (NP) and the GLP groups. In this study, the modified ethanol gastritis model was used. The rats were administrated 56% ethanol orally every other day. The dose of ethanol was 8 g/kg body weight. The normal group received the same amount of normal saline instead of ethanol. The NP and the GLP groups were treated with injection of saline and GLP respectively. The control group received no treatment. Two local acupoints CV12 (中脘) and ST36 (足三里) were used. All laboratory rats underwent treatment for 15 days. On last day, the rats were sacrificed and their stomachs were immediately excised. Results: Ulcers of the gastric mucosa appeared as elongated bands of hemorrhagic lesions parallel to the long axis of the stomach. In the NP and GLP groups, the injuries to the gastric mucosal injuries were not as severe as they were in the control group. Wound healings of the chronic gastric ulcers was promoted by using GLP and significant alterations of the indices in the gastric mucosa were observed. Such protection was demonstrated by gross appearance, histology and immunehistochemistry staining for Bcl-2-associated X (BAX), B-cell lymphoma 2 (Bcl-2) and Transforming growth factor-beta 1 (TGF-${\beta}1$). Conclusion: These results suggest that GLP at CV12 and ST36 can provide significant protection to the gastric mucosa against an ethanol induced chronic gastric ulcer.