• Title/Summary/Keyword: RNA degradation

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Transcription factor EGR-1 transactivates the MMP1 gene promoter in response to TNFα in HaCaT keratinocytes

  • Yeo, Hyunjin;Lee, Jeong Yeon;Kim, JuHwan;Ahn, Sung Shin;Jeong, Jeong You;Choi, Ji Hye;Lee, Young Han;Shin, Soon Young
    • BMB Reports
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    • v.53 no.6
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    • pp.323-328
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    • 2020
  • Matrix metalloproteinase 1 (MMP-1), a calcium-dependent zinccontaining collagenase, is involved in the initial degradation of native fibrillar collagen. Tissue necrosis factor-alpha (TNFα) is a pro-inflammatory cytokine that is rapidly produced by dermal fibroblasts, monocytes/macrophages, and keratinocytes and regulates inflammation and damaged-tissue remodeling. MMP-1 is induced by TNFα and plays a critical role in tissue remodeling and skin aging processes. However, the regulation of the MMP1 gene by TNFα is not fully understood. We aimed to find additional cis-acting elements involved in the regulation of TNFα-induced MMP1 gene transcription in addition to the nuclear factor-kappa B (NF-κB) and activator protein 1 (AP1) sites. Assessments of the 5'-regulatory region of the MMP1 gene, using a series of deletion constructs, revealed the requirement of the early growth response protein 1 (EGR-1)-binding sequence (EBS) in the proximal region for proper transcription by TNFα. Ectopic expression of EGR-1, a zinc-finger transcription factor that binds to G-C rich sequences, stimulated MMP1 promoter activity. The silencing of EGR-1 by RNA interference reduced TNFα-induced MMP-1 expression. EGR-1 directly binds to the proximal region and transactivates the MMP1 gene promoter. Mutation of the EBS within the MMP1 promoter abolished EGR-1-mediated MMP-1 promoter activation. These data suggest that EGR-1 is required for TNFα-induced MMP1 transcriptional activation. In addition, we found that all three MAPKs, ERK1/2, JNK, and p38 kinase, mediate TNFα-induced MMP-1 expression via EGR-1 upregulation. These results suggest that EGR-1 may represent a good target for the development of pharmaceutical agents to reduce inflammation-induced MMP-1 expression.

Cell Biological Study on Factors Affecting Brain Formation at Early Chick Embryo (1) The Effect of Serotonin (초기 계배의 뇌형성에 미치는 몇가지 요인에 관한 세포 생물학적 연구 (1) Serotonin의 영향)

  • 최임순;주상옥;주충노;오억수;신길상
    • The Korean Journal of Zoology
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    • v.32 no.1
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    • pp.55-73
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    • 1989
  • The effect of tryptophan or serotonin on the early stage of chick brain development has been morphologically investigated using an electron microscope. The electron micrographs of neural plate cells of 1-day chick embryo treated with tryptophan or serotonin showed irregularity, evagination and disruption of nuclear membrane and nuclear chromatin condenstation, nucleolar margination and segregation. Hypertrophy of stalks, vesicles and vaculoes were seen and dilated and disrupted rough endoplasmic reticulum and underdeveloped neurotubules were also observed. In mesenchyme cells of tryptophan or serotonin administered 18 hr embryo, irregular nuclear membrane, swollen mitochondria, dilated rough endoplasmic reticulum and very large yolk granules were observed. Furthermore, DNA, RNA and protein contents of the embryos treated with typtophan or serotonin were considerably lower than those of control group. The amount of tubulin of the experimental groups was also greatly lower than that of control, suggesting that the impairment of microtubule formation occurred. Tryptophan or serotonin administration might depress the biosynthesis, of nucleic acid and protein including some enzymes tested. It seems that the serotonin formed from exogeneous tryptophan might inhibit the degradation of yolk granule by feedback regulation mechanism so as to impair microtububle and microvilli formation followed by a malformation of chick embryos.

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The Effects of Somatid on the Cytotoxicity of Cancer Cells and Human Papillomavirus Type 16 E6 and E7 Oncogenes (생기액(生肌液)의 세포독성 및 자궁경부암 바이러스 (HPV 16 type) 암 유발인자 E6와 E7의 작용에 미치는 효과)

  • Joung, Ok;Cho, Young-Sik;Cho, Cheong-Weon;Lee, Kyung-Ae;Shim, Jung-Hyun;Cho, Min-Chul;Lee, Hong-Soo;Yeom, Young-Il;Kim, Sang-Bom;Park, Sue-Nie;Yoon, Do-Young
    • YAKHAK HOEJI
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    • v.44 no.4
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    • pp.340-346
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    • 2000
  • Cervical cancer is one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papilloma viruses (HPV types 16 and 18) and cervical cancer has been well known. An extract of natural products, named as Somatid, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. This Somatid inhibited the proliferation of human cervical cancer cell lines (C-33A, SiHa, CaSki) and HaCaT keratinocytes in a dose response manner, In vitro binding assay and ELISA showed that Somatid inhibited the in vitro biding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, Somatid inhibited the in vitro binding of E7 and Rb which is essential tumor suppressor for the control of cell cycle. The levels of mRNA for E6 and E7 were also decreased by Somatid. Our data suggested that Somatid inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus can be used as a putative anti-HPV agent for the treatment of cervical carcinomas caused by HPV.

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Suppression of TNF-α-induced Inflammation by Extract from Different Parts of Moringa in HaCaT Cells (HaCaT 각질형성세포에서 TNF-α에 의하여 유도되는 염증 발현에 대한 부위별 모링가 추출물의 억제 효과)

  • Lee, Hyo-Jin;Chang, Young-Chae
    • Journal of Life Science
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    • v.22 no.9
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    • pp.1254-1260
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    • 2012
  • The moringa (Moringa oleifera Lam.) plant is used both as food and an anti-allergic agent. In this study, we investigated skin protection effects of methanol extracts from the root, seed, fruit, and leaves of moringa in HaCaT keratinocyte cells. To investigate the pharmacological potential of various moringa extracts on TNF-${\alpha}$-induced collagen degradation in HaCaT cells, we measured the activity of matrix metallopeptidase-9,2 (MMP-9,2) by zymography analysis. Our results showed that all the moringa extracts inhibit the TNF-${\alpha}$-induced enzyme activity of MMP-9. In particular, moringa root extracts significantly suppressed MMP-9 and MMP-2 in a dose-dependent manner. Next, to investigate the anti-inflammation effect of the moringa extracts, we examined cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and interleukin-6 (IL-6) expression of the extracts. The results showed that both the root extracts and the seed extracts decreased the TNF-${\alpha}$-induced expression of COX-2. In addition, the root and leaf extracts reduced the expression of IL-6. However, none of the moringa extracts affected the expression of iNOS. The results suggest that moringa root extracts down-regulate MMP-9, COX-2, and IL-6 and that the root extracts offer superior skin protection effects compared with other extracts of moringa in HaCaT cells.

Isolation and Identification of Histamine Degrading Bacteria from Kwamegi (과메기에서 histamine 분해능을 나타내는 세균의 분리 동정)

  • Kim Min-Woo;Kim Young-Man
    • Journal of Life Science
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    • v.16 no.1
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    • pp.120-125
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    • 2006
  • To isolate and identify histamine degrading bacteria from Kwamegi, bacteria were screened with restriction media containing histamine. Ten strains were selected through morphological and biochemical identification procedure followed by comparison with DNA sequence of 16 rRNA gene. And also, these strains were confirmed by the histamine degrading assay such as turbidity and enzymatic assay. The results of identification are as followings : Ewingella americana B791, Arthrobacter sp. R45S, Halomonas marisflava, Psychrobacter sp. 9B-7, Bacillus sp. LMC 21002, Psychrohacter cibarius BC-220, Bacillus megaterium KL-197 were identified showing homology of $99\%,\;95\%,\;98\%,\;99\%,\;99\%,\;99\%\;and\;98\%$, respectively. Three strains remain unidentified. Arthrobacter sp. R45S, H. marisflava, Bacillus sp. LMG 21002, B. megaterium KL-197 showed histamine degrading activity, whereas, Psychrobacter sp. 9B-7 only showed weak activity. Three unidentified strains also have histamine degrading activity. In contrast, E. american B791 and p. cibarius JG-220 did not show any significant activity of histamine degradation. The strains isolated from this study showed relatively fast growth rate and histamine degrading rate as compared to those from salted mackerel.

A Review of Experimental study on Dementia in Oriental medicine;within Oriental medicine journal since 2000 (치매에 대한 최신 실험적 연구 동향;2000년 이후 한의학 학술지를 중심으로)

  • Choi, Sung-Youl;Kim, Dae-Hyun;Kim, Sang-Tae;Kim, Tae-Heon;Kang, Hyung-Won;Lyu, Yeong-Su
    • Journal of Oriental Neuropsychiatry
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    • v.19 no.1
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    • pp.125-146
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    • 2008
  • Objectives : The purpose of this study is to suggest for the following experimental study of dementia by reviewing recent oriental medicine journals that have been published since 2000. Methods: We have investigated various types of studies in relation to dementia through 90 articles that have been published from 2000 to 2007 in recent oriental medicine journals were registered Korea research foundation. Results and Conclusions : 1. Since 2000, 88 articles in relation to dementia have been published and almost of them are herbal medicine-centered studies. Also they show a tendency to increase every year. The journal of oriental neuropsychiatry carries the highest number of studies in relation to dementia. 2. According to the experimental paper, there are 30 cases of using herb simplexes, 48 cases of herb-combined prescription, and 10 cases of other ways. Especially 7 cases of using herb-combined prescription relation to Sasang constitution are all for the Taeumin. 3. There are 85 cases of Animal and cellular experimental, 60 cases of using pathologic model induced cytotoxic activity, a case of using L-NAME, 3 cases of 192 saporin, 4 cases of ibotenic acid, 10 cases of focal cerebral ischemia, 3 cases of alcohol-administered, and one case of natural degradation. 4. Moms water maze, Radial arm maze Passive avoidance learning model were using for examining learning and memory of model animal 5. We propose that following studies of dementia are to he investigated of the applied method of using siRNA with tranceduced gene, sample preparation by water-soaking, oriental medical diagnosis, standardization of differentiating symptom and herb simplexes, building the database by classified prescriptions, and experiment model which are based on precise examining mechanism with cell line as like mouse H19-7 hippocampus, rat HT22 hippocampus, astrocyte, microglia, using the model of animals at APP, PS1, BACE, CT99/PS1, APOE4, Tau, APP/PSI/Tau

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Mitochondrial Ca2+ Uptake Relieves Palmitate-Induced Cytosolic Ca2+ Overload in MIN6 Cells

  • Ly, Luong Dai;Ly, Dat Da;Nguyen, Nhung Thi;Kim, Ji-Hee;Yoo, Heesuk;Chung, Jongkyeong;Lee, Myung-Shik;Cha, Seung-Kuy;Park, Kyu-Sang
    • Molecules and Cells
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    • v.43 no.1
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    • pp.66-75
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    • 2020
  • Saturated fatty acids contribute to β-cell dysfunction in the onset of type 2 diabetes mellitus. Cellular responses to lipotoxicity include oxidative stress, endoplasmic reticulum (ER) stress, and blockage of autophagy. Palmitate induces ER Ca2+ depletion followed by notable store-operated Ca2+ entry. Subsequent elevation of cytosolic Ca2+ can activate undesirable signaling pathways culminating in cell death. Mitochondrial Ca2+ uniporter (MCU) is the major route for Ca2+ uptake into the matrix and couples metabolism with insulin secretion. However, it has been unclear whether mitochondrial Ca2+ uptake plays a protective role or contributes to lipotoxicity. Here, we observed palmitate upregulated MCU protein expression in a mouse clonal β-cell, MIN6, under normal glucose, but not high glucose medium. Palmitate elevated baseline cytosolic Ca2+ concentration ([Ca2+]i) and reduced depolarization-triggered Ca2+ influx likely due to the inactivation of voltage-gated Ca2+ channels (VGCCs). Targeted reduction of MCU expression using RNA interference abolished mitochondrial superoxide production but exacerbated palmitate-induced [Ca2+]i overload. Consequently, MCU knockdown aggravated blockage of autophagic degradation. In contrast, co-treatment with verapamil, a VGCC inhibitor, prevented palmitate-induced basal [Ca2+]i elevation and defective [Ca2+]i transients. Extracellular Ca2+ chelation as well as VGCC inhibitors effectively rescued autophagy defects and cytotoxicity. These observations suggest enhanced mitochondrial Ca2+ uptake via MCU upregulation is a mechanism by which pancreatic β-cells are able to alleviate cytosolic Ca2+ overload and its detrimental consequences.

Characterization of Cellulase from Bacillus subtilis NSC Isolated from Soil (토양으로부터 단리한 Bacillus subtilis NSC 유래 Cellulase의 특성 규명)

  • Kim, Sang Jin;Park, Chang-Su
    • Journal of Chitin and Chitosan
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    • v.23 no.4
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    • pp.228-233
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    • 2018
  • We isolated microorganisms from soil, which is sampled at forest, Kyeonbuk, Korea, as cellulolytic microorganisms. The isolated strains were identified by analysis of 16S rRNA gene from the starins. The result, four kinds of Bacillus subtilis, one kind of Bacillus amyloliquefaciens, and one kind of Bacillus cereus were identified. Among these strains, Bacillus subtilis was selected due to its high cellulase activity and this strain was named as Bacillus subtilis CNS. The optimum pH and temperature of the cellulase from Bacillus subtilis CNS was pH 5.0 and $40^{\circ}C$, respectively. In the investigation of pH and temperature stability, the cellulase from Bacillus subtilis NSC stabled pH 4.0~6.0 range and until $40^{\circ}C$ for 30 min perfectly. In the enzyme activity for various cellulosic substrate, cellulase from Bacillus subtilis CNS showed the highest activity for CM-cellulose. And, the enzyme activities for alkali swollen cellulose, Alpha-cellulose, Sigmacell-cellulose, and Avicel were approximately 31%, 8%, 8% and 4% of activity for CM-cellulose, respectively. In the degradation of CM-cellulose, the 0.26 U/ml and 0.52 U/ml of cellulase showed 0.43 and 0.76 U/ml activity for CM-cellulose after the reaction of 120 min, respectively.

Inhibitory Effects of Tannic acid on Human Skin Fibroblast Elastase Activity (사람의 섬유아세포 엘라스타제 활성에 대한 탄닌산의 억제 효과)

  • Lee, Ju-Eun;Kim, So-Young;Kim, Su-Yeon;Oh, Mi-Hee;Yun, Hye-Young;Baek, Kwang-Jin;Kwon, Nyoun-Soo;Kim, Dong-Seok
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.3
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    • pp.217-223
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    • 2008
  • Elastin is an important component of elastic fibers in the skin. Recently, many studies have reported that elastin is also involved In inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. Elastase is a metalloproteinase which acts on degradation of elastin. It is known that elastase activity is increased by ultraviolet (UV) B radiation. Thus, Increased elastase activity could be the major reason for skin elasticity reduction and winkle formation. Tannic acid is a polyphenol found in various fruits and nuts. This molecule has a potent ability to eliminate reactive oxygen species and reactive nitrogen species. In the present study, we investigated whether tannic acid has effects on elastase activity and tropoelastin synthesis. Our results showed that tannic acid reduced elastase activity significantly in a dose-dependent manner. However, the expression of tropoelastin protein and mRNA was not significantly affected by tannic acid. From these results, we suggest that tannic acid may contribute to block tortuosity of elastic fibers by inhibiting elastase. Thus, tannic acid might be developed for a possible agent to Inhibit skin aging.

Changes in expression of the autophagy-related genes microtubule-associated protein 1 light chain 3β and autophagy related 7 in skeletal muscle of fattening Japanese Black cattle: a pilot study

  • Nakanishi, Tomonori;Tokunaga, Tadaaki;Ishida, Takafumi;Kobayashi, Ikuo;Katahama, Yuta;Yano, Azusa;Erickson, Laurie;Kawahara, Satoshi
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.4
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    • pp.592-598
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    • 2019
  • Objective: Autophagy is a bulk degradation system for intracellular proteins which contributes to skeletal muscle homeostasis, according to previous studies in humans and rodents. However, there is a lack of information on the physiological role of autophagy in the skeletal muscle of meat animals. This study was planned as a pilot study to investigate changes in expression of two major autophagy-related genes, microtubule-associated protein 1 light chain $3{\beta}$ (MAP1LC3B) and autophagy related 7 (ATG7) in fattening beef cattle, and to compare them with skeletal muscle growth. Methods: Six castrated Japanese Black cattle (initial body weight: $503{\pm}20kg$) were enrolled in this study and fattened for 7 months. Three skeletal muscles, M. longissimus, M. gluteus medius, and M. semimembranosus, were collected by needle biopsy three times during the observation period, and mRNA levels of MAP1LC3B and ATG7 were determined by quantitative reverse-transcription polymerase chain reaction. The expression levels of genes associated with the ubiquitin-proteasome system, another proteolytic mechanism, were also analyzed for comparison with autophagy-related genes. In addition, ultrasonic scanning was repeatedly performed to measure M. longissimus area as an index of muscle growth. Results: Our results showed that both MAP1LC3B and ATG7 expression increased over the observation period in all three skeletal muscles. Interestingly, the increase in expression of these two genes in M. longissimus was highly correlated with ultrasonic M. longissimus area and body weight. On the other hand, the expression of genes associated with the ubiquitin-proteasome system was unchanged during the same period. Conclusion: These findings suggest that autophagy plays an important role in the growth of skeletal muscle of fattening beef cattle and imply that autophagic activity affects meat productivity.