• 제목/요약/키워드: Real-Time Polymerase Chain Reaction

검색결과 809건 처리시간 0.026초

Effects of Gyeongshingangjeehwan 18 on Pancreatic Fibroinflammation in High-Fat Diet-Fed Obese C57BL/6J Mice

  • Jang, Joonseong;Park, Younghyun;Yoon, Michung
    • 대한의생명과학회지
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    • 제24권4호
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    • pp.341-348
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    • 2018
  • The polyherbal drug Gyeongshingangjeehwan 18 (GGEx18) from Rheum palmatum L. (Polygonaceae), Laminaria japonica Aresch (Laminariaceae), and Ephedra sinica Stapf (Ephedraceae) has traditionally been used as an antiobesity drug in Korean local clinics. This study investigates the effects of GGEx18 on pancreatic fibroinflammation in high-fat diet (HFD)-fed obese C57BL/6J mice and the molecular mechanism involved in this process. After HFD-fed obese C57BL/6J mice were treated with GGEx18 (125, 250, and 500 mg/kg) for 12 weeks, variables and determinants of obesity, pancreatic inflammation, and fibrosis were measured using histology, immunohistochemistry, and real-time polymerase chain reaction. Administration of GGEx18 at 500 mg/kg/day to obese mice decreased body weight gain, mesenteric adipose tissue mass, and adipocyte size. GGEx18 treatment not only reduced mast cells and CD68-immunoreactive cells, but also decreased collagen levels and ${\alpha}$-smooth muscle actin-positive cells in the pancreas of HFD-fed mice. Concomitantly, GGEx18 decreased the expression of genes for inflammation (i.e., CD68 and tumor necrosis factor ${\alpha}$) and fibrosis (i.e., collagen ${\alpha}1$ and transforming growth factor ${\beta}$) in the pancreas of obese mice. These results suggest that GGEx18 may inhibit visceral obesity and related pancreatic fibroinflammation in HFD-fed obese mice.

Gallic acid caused cultured mice TM4 Sertoli cells apoptosis and necrosis

  • Li, Wanhong;Yue, Xiangpeng;Li, Fadi
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권5호
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    • pp.629-636
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    • 2019
  • Objective: The study was designed to determine the cytotoxic effect of gallic acid (GA), obtained by the hydrolysis of tannins, on mice TM4 Sertoli cells apoptosis. Methods: In the present study, non-tumorigenic mice TM4 Sertoli cells were treated with different concentrations of GA for 24 h. After treatment, cell viability was evaluated using WST-1, mitochondrial dysfunction, cells apoptosis and necrosis was detected using JC-1, Hoechst 33342 and propidium iodide staining. The expression levels of Cyclin B1, proliferating cell nuclear antigen (PCNA), Bcl-2-associated X protein (BAX), and Caspase-3 were also detected by quantitative real-time polymerase chain reaction and Western-blotting. Results: The results showed that 20 to $400{\mu}M$ GA inhibited viability of TM4 Sertoli cells in a dose-dependent manner. Treatment with $400{\mu}M$ GA significantly inhibited PCNA and Cyclin B1 expression, however up-regulated BAX and Caspase-3 expression, caused mitochondrial membrane depolarization, activated Caspase-3, and induced DNA damage, thus, markedly increased the numbers of dead cells. Conclusion: Our findings showed that GA could disrupt mitochondrial function and caused TM4 cells to undergo apoptosis and necrosis.

Knockdown of Circ_0000144 Suppresses Cell Proliferation, Migration and Invasion in Gastric Cancer Via Sponging MiR-217

  • Ji, Fengcun;Lang, Chao;Gao, Pengfei;Sun, Huanle
    • Journal of Microbiology and Biotechnology
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    • 제31권6호
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    • pp.784-793
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    • 2021
  • Previous studies have uncovered the role of circ_0000144 in various tumors. Here, we investigated the function and mechanism of circ_0000144 in gastric cancer (GC) progression. The expression of circ_0000144 in GC tissues and cells was detected through quantitative real-time polymerase chain reaction (qRT-PCR) method. Gain- and loss-of-function experiments including colony formation, wound healing and transwell assays were performed to examine the role of circ_0000144 in GC cells. Furthermore, western blot was conducted to determine the expressions of epithelial mesenchymal transition (EMT)-related proteins. The interaction between circ_0000144 and miR-217 was analyzed by bioinformatic analysis and luciferase reporter assays. The circ_0000144 expression was obviously upregulated in GC tissues and cells. Silencing of circ_0000144 inhibited cell proliferation, migration and invasion of GC cells, but ectopic expression of circ_0000144 showed the opposite results. Moreover, circ_0000144 sponged miR-217, and rescue assays revealed that silencing miR-217 expression reversed the inhibitory effect of circ_0000144 knockdown on the progress of GC. Our findings reveal that circ_0000144 inhibition suppresses GC cell proliferation, migration and invasion via absorbing miR-217, providing a new biomarker and potential therapeutic target for treatment of GC.

Improved immune-enhancing activity of egg white protein ovotransferrin after enzyme hydrolysis

  • Lee, Jae Hoon;Kim, Hyeon Joong;Ahn, Dong Uk;Paik, Hyun-Dong
    • Journal of Animal Science and Technology
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    • 제63권5호
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    • pp.1159-1168
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    • 2021
  • Ovotransferrin (OTF), an egg protein known as transferrin family protein, possess strong antimicrobial and antioxidant activity. This is because OTF has two iron binding sites, so it has a strong metal chelating ability. The present study aimed to evaluate the improved immune-enhancing activities of OTF hydrolysates produced using bromelain, pancreatin, and papain. The effects of OTF hydrolysates on the production and secretion of pro-inflammatory mediators in RAW 264.7 macrophages were confirmed. The production of nitric oxide (NO) was evaluated using Griess reagent and the expression of inducible nitric oxide synthase (iNOS) were evaluated using quantitative real-time polymerase chain reaction (PCR). And the production of pro-inflammatory cytokines (tumor necrosis factor [TNF]-α and interleukin [IL]-6) and the phagocytic activity of macrophages were evaluated using an ELISA assay and neutral red uptake assay, respectively. All OTF hydrolysates enhanced NO production by increasing iNOS mRNA expression. Treating RAW 264.7 macrophages with OTF hydrolysates increased the production of pro-inflammatory cytokines and the phagocytic activity. The production of NO and pro-inflammatory cytokines induced by OTF hydrolysates was inhibited by the addition of specific mitogen-activated protein kinase (MAPK) inhibitors. In conclusion, results indicated that all OTF hydrolysates activated RAW 264.7 macrophages by activating MAPK signaling pathway.

Conjugation of vascular endothelial growth factor to poly lactic-co-glycolic acid nanospheres enhances differentiation of embryonic stem cells to lymphatic endothelial cells

  • Yoo, Hyunjin;Choi, Dongyoon;Choi, Youngsok
    • Animal Bioscience
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    • 제34권4호
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    • pp.533-538
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    • 2021
  • Objective: Pluripotent stem cell-derived lymphatic endothelial cells (LECs) show great promise in their therapeutic application in the field of regenerative medicine related to lymphatic vessels. We tested the approach of forced differentiation of mouse embryonal stem cells into LECs using biodegradable poly lactic-co-glycolic acid (PLGA) nanospheres in conjugation with growth factors (vascular endothelial growth factors [VEGF-A and VEGF-C]). Methods: We evaluated the practical use of heparin-conjugated PLGA nanoparticles (molecular weight ~15,000) in conjugation with VEGF-A/C, embryoid body (EB) formation, and LEC differentiation using immunofluorescence staining followed by quantification and quantitative real-time polymerase chain reaction analysis. Results: We showed that formation and differentiation of EB with VEGF-A/C-conjugated PLGA nanospheres, compared to direct supplementation of VEGF-A/C to the EB differentiation media, greatly improved yield of LYVE1(+) LECs. Our analyses revealed that the enhanced potential of LEC differentiation using VEGF-A/C-conjugated PLGA nanospheres was mediated by elevation of expression of the genes that are important for lymphatic vessel formation. Conclusion: Together, we not only established an improved protocol for LEC differentiation using PLGA nanospheres but also provided a platform technology for the mechanistic study of LEC development in mammals.

Human Mastadenovirus Infections and Meteorological Factors in Cheonan, Korea

  • Oh, Eun Ju;Park, Joowon;Kim, Jae Kyung
    • 한국미생물·생명공학회지
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    • 제49권2호
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    • pp.249-254
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    • 2021
  • The study of the impact of weather on viral respiratory infections enables the assignment of causality to disease outbreaks caused by climatic factors. A better understanding of the seasonal distribution of viruses may facilitate the development of potential treatment approaches and effective preventive strategies for respiratory viral infections. We analyzed the incidence of human mastadenovirus infection using real-time reverse transcription polymerase chain reaction in 9,010 test samples obtained from Cheonan, South Korea, and simultaneously collected the weather data from January 1, 2012, to December 31, 2018. We used the data collected on the infection frequency to detect seasonal patterns of human mastadenovirus prevalence, which were directly compared with local weather data obtained over the same period. Descriptive statistical analysis, frequency analysis, t-test, and binomial logistic regression analysis were performed to examine the relationship between weather, particulate matter, and human mastadenovirus infections. Patients under 10 years of age showed the highest mastadenovirus infection rates (89.78%) at an average monthly temperature of 18.2℃. Moreover, we observed a negative correlation between human mastadenovirus infection and temperature, wind chill, and air pressure. The obtained results indicate that climatic factors affect the rate of human mastadenovirus infection. Therefore, it may be possible to predict the instance when preventive strategies would yield the most effective results.

Astaxanthin Ameliorates Atopic Dermatitis by Inhibiting the Expression of Signal Molecule NF-kB and Inflammatory Genes in Mice

  • Donghwan, Kim;Yong-Suk, Kim;Ho Sueb, Song
    • Journal of Acupuncture Research
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    • 제39권4호
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    • pp.304-309
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    • 2022
  • Background: This study was conducted to determine the anti-inflammatory effect of astaxanthin, on atopic dermatitis. Methods: Changes in mouse body weight, lymph node weight, and the degree of improvement in symptoms were measured to determine the inflammatory response. Real-time reverse transcription-polymerase chain reaction tests were performed to determine the degree of expression of inflammation-related cytokines (IL-31 and IL-33 and chemokines such as CCL17 and CCL22), and western blot analysis was performed to evaluate the expression of inflammation-related factors (iNOS, COX-2, and NF-kB signaling molecules p-IkBα, p50, p-65 and pSTAT3). Results: The degree of symptoms significantly improved in the PA+AX group. Lymph node weight in the PA+AX group was lower than the PA group. Inflammatory cytokines (IL-31, IL-33, and inflammatory chemokines such as CCL17 and CCL22) were significantly reduced in the PA+AX group compared with the PA group. The expression of inflammatory genes (iNOS, COX-2, NF-kB and signaling molecules (p-IkBα, p50, p65, and p-STAT 3) was lower in the PA+AX group compared with the PA group. Conclusion: Astaxanthin may modulate the inflammatory response in a mouse model of atopic dermatitis and has an anti-inflammatory effect.

Hangambujeongsan or Kangai Fuzheng Powder shows the anti-cancer effect by enhancing macrophage activation

  • Yang, Wan-Quan;Han, Hyung Soo
    • 대한본초학회지
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    • 제29권1호
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    • pp.1-6
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    • 2014
  • Objectives : Many of currently used anti-cancer drugs were developed to target cell death mechanisms and had serious side effects by causing damage to normal cells. Hangambujeongsan or Kangai Fuzheng Powder was a mixture based on the traditional Chinese medicine. It had been used in the local Chinese hospitals to treat cancer patients for decades and had shown a certain level of beneficial effects without major toxic effects. But its mechanism of action had not been elucidated yet. Thus this study aimed to investigate the effects of Kangai Fuzheng Powder in an in vitro experiment. Methods : Cancer lines or RAW264.7 mouse macrophage cells were treated with Kangai Fuzheng Powder. Cell viability was measured by MTT assay, and morphological observation was also performed. Gene expression of cytokines in macrophages was determined by real-time polymerase chain reaction. Phagocytic function assay was also performed in macrophage cells. Results : Kangai Fuzheng Powder had no direct detrimental effect on cancer cells. When macrophages were co-cultured with cancer cells, Kangai Fuzheng Powder had toxic effect on cancer cells. After exposing macrophages to Kangai Fuzheng Powder, macrophages transformed into activated form and the mRNA level of tumor necrosis factor-alpha, interleukin-1beta, interleukin-6, interleukin-10 and monocyte chemotactic protein-1 was significantly enhanced. Phagocytic activity of macrophages was dramatically potentiated. Conclusions : We demonstrated that anti-cancer effect of Kangai Fuzheng Powder was related to activation of macrophages including enhanced cytokine production and phagocytic function.

Effects of red ginseng extract on gut microbial distribution

  • Kim, Young Kyun;Yum, Keun-Sang
    • Journal of Ginseng Research
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    • 제46권1호
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    • pp.91-103
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    • 2022
  • Background: Red ginseng extract boosts immunity against inflammation and cancer in the human body. However, studies on the effects of red ginseng extract on the gut microbiome remain unexplored. Methods: In 2019, the positive effects and changes in the gut microbiome after administering 1 pack (3 g) of red ginseng extract per day to 53 adults aged 40 to 75 for 24 weeks were investigated. The gut microbial environment changes were qualitatively and quantitatively analyzed using next-generation sequencing and real-time polymerase chain reaction technology. Results: On comparing and analyzing alpha diversity and beta diversity, the microbial pattern showed significant differences (OTUs p = 0.003, chao1 p < 0.001, Bray-Curtis p = 0.001) before and after ingestion of red ginseng extract, indicating that gut microbial richness increased after ingestion. Moreover, after comparing and analyzing the gut microbiome's differences after red ginseng extract intake, significant differences were noted between three strains at the phylum level and among 57 strains at the genus level. Conclusion: This study proposes the potential use of red ginseng extract as a prebiotic after confirming its positive effects, including increasing gut microbiome richness, reducing harm to the gut microbiome, and increasing the number of some strains in the gut microbiome.

요양보호시설 이용중인 알츠하이머 치매환자의 구강 병원성 미생물 분석 (Analysis of oral pathogenic microorganisms in Alzheimer's dementia patients using nursing facilities)

  • 정서윤;정미애;김춘성;김수관
    • 한국치위생학회지
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    • 제22권5호
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    • pp.411-416
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    • 2022
  • Objectives: This study aimed to identify pathogenic microorganisms in the oral cavity of Alzheimer's dementia patients and recognize the necessity and importance of oral prevention management. Methods: The participants comprised 40 dementia patients aged 60 years or older and general patients who were using nursing care facilities in Gwangju from February to July 2017. Samples were collected with Eazyperio products for oral pathogenic microbial testing. Eighteen types of bacteria could be detected by analyzing Multiplex-Quantity Real Time polymerase chain reaction at a genetic testing agency. Results: The study comprised more women than men. Most participants were in their 80s. Statistically significant differences were observed in some oral pathogenic microorganisms. Conclusions: Pathogenic microorganisms could more easily proliferate in the oral cavities of Alzheimer's dementia patients than they could among general elderly participants due to a lack of awareness of oral hygiene and prevention management. To improve this, it is considered necessary to deploy oral health care professionals.