scholarly journals The Role of Coconut Oil to Increase Expression of MMP-9, PDGF-BB, and TGF-β1 in NIH-3T3 Cell Line

2019 ◽  
Vol 7 (22) ◽  
pp. 3733-3736
Author(s):  
Dian Ika Perbina Meliala ◽  
Jansen Silalahi ◽  
Yuandani Yuandani ◽  
Linda Margata ◽  
Denny Satria
Keyword(s):  
Healing Process ◽  
Coconut Oil ◽  
Wound Healing Process ◽  
3T3 Cells ◽  
Virgin Coconut Oil ◽  
Nih3t3 Cells ◽  
Nih 3T3 ◽  
Tgf Β1 ◽  
Nih 3T3 Cells

AIM: The objective of the study was to evaluate protein expression in NIH 3T3 cells that are treated with virgin coconut oil (VCO) and hydrolysed of virgin coconut oil (HVCO) in vitro. METHODS: Coconut oil used in this study was virgin coconut oil (VCO) and VCO hydrolysed by Rhizomucor miehei (HVCO). NIH 3T3 cells (5x105 cells/well) were seeded in nine wells and incubated for overnight, then divided into three groups. Each group consisted of three wells. Group one without treatment, group two added VCO, and group three added HVCO and then incubated for overnight. One well in each group was added MMP-9, PDGF-BB, and TGF-β1 and incubated one hour. Finally, expressions of MMP-9, PDGF-BB, and TGF-β1 were detected using immunocytochemistry method. RESULTS: The results of the study showed that VCO and HVCO increased protein expressions of MMP-9, PDGF-BB, and TGF-β1. Percentage of MMP-9 expressions treated by VCO increased from 2.89 ± 0.07 to 28.16 ± 0.34, PDGF-BB from 28.11 ± 0.13 to 48.53 ± 0.49, and TGF-β1 from 4.19 ± 0.08 to 18.41 ± 0.54. Percentage of MMP-9 expressions treated by HVCO increased from 2.89 ± 0.07 to 55.40 ± 0.94, PDGF-BB from 28.11 ± 0.13 to 61.65 ± 0.42, and TGF-β1 from 4.19 ± 0.08 to 36.35 ± 0.67. CONCLUSION: VCO and HVCO increase the expression of MMP-9, PDGF-BB, dan TGF-β1 in NIH3T3 cells and therefore, coconut oil active in the wound healing process. HVCO is more than active than VCO.

scholarly journals The Activity of Hydrolyzed Virgin Coconut Oil to Increase Proliferation and Cyclooxygenase-2 Expression towards on NIH 3T3 Cell Line in Wound Healing Process

2019 ◽  
Vol 7 (19) ◽  
pp. 3164-3168
Author(s):  
Jansen Silalahi ◽  
Yuandani Yuandani ◽  
Dian Ika Perbina Br Meliala ◽  
Linda Margata ◽  
Denny Satria
Keyword(s):  
Fatty Acids ◽  
Cell Proliferation ◽  
Wound Healing ◽  
Healing Process ◽  
Coconut Oil ◽  
Acid Value ◽  
Wound Healing Process ◽  
Virgin Coconut Oil ◽  
Cox 2 ◽  
Nih 3T3

AIM: This study aims to determine the effect of hydrolysed virgin coconut oil (HVCO) to increase cell proliferation, COX-2 expression of NIH 3T3. METHODS: The sample used was Virgin Coconut Oil (VCO). VCO was partially hydrolysed using lipase from Rhizomucor miehei (active on sn-1,3 position) to produce hydrolysed VCO (HVCO) composed of free fatty acids, 2-monoglycerides. Then acid value was determined. The effect of HVCO on proliferation was evaluated using the MTT method. Wound healing assay was established by a cell migration method, and COX-2 expression was determined using RT-PCR. RESULTS: Acid value is 135.89 ± 0.12 mg NaOH/g oil and free fatty acids (FFA) is 48.50 ± 0.06%. The effect of HVCO 62.5 µg/mL on cell proliferation after 24h, 48h, and 72h incubation found as viable cells are 109.24 ± 0.52%; 118.26 ± 0.91% and 106.59 ± 0.74%. Percent of wound closed after 24 h and 48 h incubation are 69.94 ± 0.54% and 100.00 ± 0.00%, and expression of COX-2 increased from 1 (control) to 1.83 (HVCO). CONCLUSION: The results suggest that HVCO is effective to increase cells proliferation and hence wound healing process.

scholarly journals Endoplasmic Reticulum Stress Is a Determinant of Retrovirus-Induced Spongiform Neurodegeneration

2003 ◽  
Vol 77 (23) ◽  
pp. 12617-12629 ◽  
Author(s):  
Derek E. Dimcheff ◽  
Srdjan Askovic ◽  
Audrey H. Baker ◽  
Cedar Johnson-Fowler ◽  
John L. Portis
Keyword(s):  
Endoplasmic Reticulum ◽  
Transmissible Spongiform Encephalopathy ◽  
Viral Envelope ◽  
Spongiform Encephalopathy ◽  
Envelope Gene ◽  
Viral Envelope Protein ◽  
3T3 Cells ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

ABSTRACT FrCasE is a mouse retrovirus that causes a fatal noninflammatory spongiform neurodegenerative disease with pathological features strikingly similar to those induced by transmissible spongiform encephalopathy (TSE) agents. Neurovirulence is determined by the sequence of the viral envelope protein, though the specific role of this protein in disease pathogenesis is not known. In the present study, we compared host gene expression in the brain stems of mice infected with either FrCasE or the avirulent virus F43, differing from FrCasE in the sequence of the envelope gene. Four of the 12 disease-specific transcripts up-regulated during the preclinical period represent responses linked to the accumulation of unfolded proteins in the endoplasmic reticulum (ER). Among these genes was CHOP/GADD153, which is induced in response to conditions that perturb endoplasmic reticulum function. In vitro studies with NIH 3T3 cells revealed up-regulation of CHOP as well as BiP, calreticulin, and Grp58/ERp57 in cells infected with FrCasE but not with F43. Immunoblot analysis of infected NIH 3T3 cells demonstrated the accumulation of uncleaved envelope precursor protein in FrCasE- but not F43-infected cells, consistent with ER retention. These results suggest that retrovirus-induced spongiform neurodegeneration represents a protein-folding disease and thus may provide a useful tool for exploring the causal link between protein misfolding and the cytopathology that it causes.

scholarly journals Transformation of NIH 3T3 cells by cotransfection with c-src and nuclear oncogenes.

1987 ◽  
Vol 7 (10) ◽  
pp. 3582-3590 ◽  
Author(s):  
D Shalloway ◽  
P J Johnson ◽  
E O Freed ◽  
D Coulter ◽  
W A Flood
Keyword(s):  
3T3 Cells ◽  
Focus Formation ◽  
Adenovirus E1a ◽  
Rous Sarcoma ◽  
Cellular Homolog ◽  
Sarcoma Virus ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

pp60c-src, the cellular homolog of the Rous sarcoma virus transforming protein, does not completely transform cells even when present at high levels, but has been shown to be involved in polyomavirus-induced transformation when activated by polyomavirus middle T (pmt)-antigen binding. Here we show that cotransfection, but not solo transfection, of expression plasmids for c-src and either adenovirus E1A, v-myc, c-myc, or the 5' half of polyomavirus large T (pltN) antigen into NIH 3T3 cells induces anchorage-independent growth, enhanced focus formation, and, for pltN cotransfection, tumorigenicity in adult NFS mice. Enhancement of transformation was not observed with polyomavirus small t (pst) antigen. Cotransfection of c-src with pltN induced modification of pp60c-src that altered its electrophoretic mobility and in vivo phosphorylation state and stimulated its in vitro kinase activity. Similar alterations were not seen after c-src-E1A cotransfection, suggesting that at least two different mechanisms of enhancement are involved.

Activation of mitogen-activated protein kinase kinase by v-Raf in NIH 3T3 cells and in vitro

Science ◽  
1992 ◽  
Vol 257 (5075) ◽  
pp. 1404-1407 ◽  
Author(s):  
P Dent ◽  
W Haser ◽  
T. Haystead ◽  
L. Vincent ◽  
T. Roberts ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Mitogen Activated Protein Kinase ◽  
3T3 Cells ◽  
Mitogen Activated Protein ◽  
Nih 3T3 ◽  
Protein Kinase Kinase ◽  
Nih 3T3 Cells

scholarly journals Activation of Ras in vitro and in intact fibroblasts by the Vav guanine nucleotide exchange protein.

1994 ◽  
Vol 14 (2) ◽  
pp. 906-913 ◽  
Author(s):  
E Gulbins ◽  
K M Coggeshall ◽  
C Langlet ◽  
G Baier ◽  
N Bonnefoy-Berard ◽  
...  
Keyword(s):  
Map Kinases ◽  
Guanine Nucleotide ◽  
Nucleotide Exchange ◽  
3T3 Cells ◽  
Transfected Cells ◽  
Guanine Nucleotide Exchange ◽  
Nih 3T3 ◽  
Exchange Activity ◽  
Nih 3T3 Cells

We recently identified Vav, the product of the vav proto-oncogene, as a guanine nucleotide exchange factor (GEF) for Ras. Vav is enzymatically activated by lymphocyte antigen receptor-coupled protein tyrosine kinases or independently by diglycerides. To further evaluate the physiological role of Vav, we assessed its GDP-GTP exchange activity against several Ras-related proteins in vitro and determined whether Vav activation in transfected NIH 3T3 fibroblasts correlates with the activity status of Ras and mitogen-activated protein (MAP) kinases. In vitro translated purified Vav activated by phorbol myristate acetate (PMA) or phosphorylation with recombinant p56lck displayed GEF activity against Ras but not against recombinant RacI, RacII, Ral, or RhoA proteins. Expression of vav or proto-vav in stably transfected NIH 3T3 cells led to a approximately 10-fold increase in basal or PMA-stimulated Ras exchange activity, respectively, in total-cell lysates and Vav immunoprecipitates. Elevated GEF activity was paralleled in each case by a significant increase in the proportion of active, GTP-bound Ras. PMA had a minimal effect on the low Ras. GTP level in untransfected control fibroblasts but increased it from 20 to 37% in proto-vav-transfected cells. vav-transfected cells displayed a constitutively elevated Ras. GTP level (35%), which was not increased further by PMA treatment. MAP kinases, known downstream intermediates in Ras-dependent signaling pathways, similarly exhibited increased basal or PMA-stimulated activity in Vav-expressing cells by comparison with normal NIH 3T3 cells. These results demonstrate a physiologic interaction between Vav and its target, Ras, leading to MAP kinase activation.

scholarly journals Topical ozonated virgin coconut oil improves wound healing and increases HSP90α, VEGF-A, EGF, bFGF, and CD34 in diabetic ulcer mouse model of wound healing

F1000Research ◽  
2021 ◽  
Vol 9 ◽  
pp. 580
Author(s):  
Renni Yuniati ◽  
Prasetyowati Subchan ◽  
Wibi Riawan ◽  
Matthew Brian Khrisna ◽  
Maryam Restiwijaya ◽  
...  
Keyword(s):  
Wound Healing ◽  
Mouse Model ◽  
Healing Process ◽  
Coconut Oil ◽  
Control Group ◽  
Wound Healing Process ◽  
Virgin Coconut Oil ◽  
Diabetic Ulcer ◽  
Macroscopic Appearance ◽  
Wound Length

Background: Diabetes is a disease that affects people worldwide, including in Indonesia. The prevalence of diabetes in Indonesia is increasing from year to year. One of the most devastating complications of diabetes mellitus is diabetic ulcers, which is a limb-threatening complication. Over the past few decades, ozone generated using plasma medical technology has been investigated as an agent that helps wound healing. This study aims to evaluate the effects of topical ozonated virgin coconut oil (VCO) in a diabetic wound mouse model. Methods: This study was an experimental study with a post-test control design. An ulcer wound model was made in 50 diabetic male Wistar mice, divided into five groups, and a control group of 10 non-diabetic mice. The control groups were given conventional therapy only and the treatment groups were also given topical ozonated VCO with different flow durations (0 min, 90 min, 7 h, 14 h). Macroscopic appearance and wound contraction were observed. HSP90β, VEGF-A, EGF, bFGF, and CD34 levels were measured from the immunostained slices of wound margins. Results: The reduction of wound length was proportionally related to the duration of ozone flow. Ozonated VCO with a longer duration of ozone flow healed the wound more quickly and had the shortest wound length. VCO with ozone flow for 14 hours (16837.10 µm) had the biggest reduction in wound length compared to other groups. The wounds treated with ozonated VCO showed an increase in HSP90β, VEGF-A, EGF, bFGF, and CD34 levels that correlated to improved wound healing. A longer period of treatment resulted in higher levels of wound healing biomarkers compared to shorter therapeutic durations. Conclusions: Topical ozonated VCO improved the wound healing process in a diabetic ulcer mouse model by improving macroscopic wound appearance and increasing levels of wound healing biomarkers.

scholarly journals Topical ozonated virgin coconut oil improves wound healing and increases HSP90α, VEGF-A, EGF, bFGF and CD34 in diabetic ulcer mouse model of wound healing

F1000Research ◽  
2020 ◽  
Vol 9 ◽  
pp. 580
Author(s):  
Renni Yuniati ◽  
Prasetyowati Subchan ◽  
Wibi Riawan ◽  
Matthew Brian Khrisna ◽  
Maryam Restiwijaya ◽  
...  
Keyword(s):  
Wound Healing ◽  
Mouse Model ◽  
Healing Process ◽  
Coconut Oil ◽  
Control Group ◽  
Wound Healing Process ◽  
Virgin Coconut Oil ◽  
Diabetic Ulcer ◽  
Macroscopic Appearance ◽  
Wound Length

Background: Diabetes is a disease that affects people worldwide, including in Indonesia. The prevalence of diabetes in Indonesia is increasing from year to year. One of the most devastating complications of diabetes mellitus is diabetic ulcers, which is a limb-threatening complication. Over the past few decades, ozone generated using plasma medical technology has been investigated as an agent that helps wound healing. This study aims to evaluate the effects of topical ozonated virgin coconut oil (VCO) in a diabetic wound mouse model. Methods: This study was an experimental study with a post-test control design. An ulcer wound model was made in 50 diabetic male Wistar mice, divided into five groups, and a control group of 10 non-diabetic mice. The control groups were given conventional therapy only and the treatment groups were also given topical ozonated VCO with different flow durations (0 min, 90 min, 7 h, 14 h). Macroscopic appearance and wound contraction were observed. HSP90β, VEGF-A, EGF, bFGF and CD34 levels were measured from the immunostained slices of wound margins. Results: The reduction of wound length was proportionally related to the duration of ozone flow. Ozonated VCO with a longer duration of ozone flow healed the wound more quickly and had the shortest wound length. VCO with ozone flow for 14 hours (16837.10 µm) had the biggest reduction in wound length compared to other groups. The wounds treated with ozonated VCO showed an increase in HSP90β, VEGF-A, EGF, bFGF and CD34 levels that correlated to improved wound healing. A longer period of treatment resulted in higher levels of wound healing biomarkers compared to shorter therapeutic durations. Conclusions: Topical ozonated VCO improved the wound healing process in a diabetic ulcer mouse model by improving macroscopic wound appearance and increasing levels of wound healing biomarkers.

Nickel-refining dust regulates the expression of inflammatory factors in NIH/3T3 cells

2019 ◽  
Vol 35 (3) ◽  
pp. 239-247 ◽  
Author(s):  
Runan Qin ◽  
Yue Wang ◽  
Shengyuan Wang ◽  
Bing Xia ◽  
Rui Xin ◽  
...  
Keyword(s):  
Inflammatory Factors ◽  
Dependent Manner ◽  
3T3 Cells ◽  
Related Factors ◽  
Experimental Conditions ◽  
Tnf Α ◽  
Cox 2 ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

Nickel (Ni) is a metal known to be a human carcinogen that occupational workers can be exposed to during the process of Ni refining. We investigated the molecular mechanism of inflammation that is induced by Ni-refining dust in a factory, using concentrations of 0, 25, 50, and 100 µg/mL for 24 h and 48 h, in vitro. Quantitative real-time polymerase chain reactions (qRT-PCR), Western blot analysis, and enzyme-linked immunosorbent assays (ELISA) were used to detect the transcriptional expression levels of nuclear factor-κB (NF-κB), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Results showed that Ni-refining dust decreased the secretion of IL-6 under the experimental conditions. In contrast, Ni-refining dust activated NF-κB expression and stimulated the secretion of TNF-α, IL-1β, iNOS, and COX-2 in a dose- and time-dependent manner. To summarize, we demonstrated that exposure to Ni-refining dust can induce the expression of NF-κB in NIH/3T3 cells and the secretion of inflammation related factors. This provides a new basis for further study of the inflammatory effects of Ni-refining dust.

FoxF1 is Required for Ciliogenesis and Distribution of Sonic Hedgehog Signaling Components in Cilium

2019 ◽  
Vol 19 (5) ◽  
pp. 326-334
Author(s):  
Lu Huang ◽  
Marco Tjakra ◽  
Desha Luo ◽  
Lin Wen ◽  
Daoxi Lei ◽  
...  
Keyword(s):  
Sonic Hedgehog ◽  
Hedgehog Signaling ◽  
3T3 Cells ◽  
Shh Signaling ◽  
Nih 3T3 ◽  
Signaling Components ◽  
Nih 3T3 Cells

Background: In vertebrates, cilium is crucial for Hedgehog signaling transduction. Forkhead box transcriptional factor FoxF1 is reported to be associated with Sonic Hedgehog (Shh) signaling in many cases. However, the role of FoxF1 in cilium remains unknown. Here, we showed an essential role of FoxF1 in the regulation of ciliogenesis and in the distribution of Shh signaling components in cilium. Methods: NIH/3T3 cells were serum starved for 24h to induce cilium. Meanwhile, shRNA was used to knockdown the FoxF1 expression in the cells and CRISPR/Cas9 was used to generate the FoxF1 zebrafish mutant. The mRNA and protein expression of indicated genes were detected by the qRT-PCR and western blot, respectively. Immunofluorescence staining was performed to detect the cilium and Shh components distribution. Results: FoxF1 knockdown decreased the cilium length in NIH/3T3 cells. Meanwhile, the disruption of FoxF1 function inhibited the expression of cilium-related genes and caused an abnormal distribution of Shh components in the cilium. Furthermore, homozygous FoxF1 mutants exhibited defective development of pronephric cilium in early zebrafish embryos. Conclusion: Together, our data illustrated that FoxF1 is required for ciliogenesis in vitro and in vivo and for the proper localization of Shh signaling components in cilium.

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