scholarly journals Unmethylated Cytosine phosphate guanine DNA (CpG-DNA) exacerbates liver Kupffer cells (KCs) inflammation through Toll-like receptor 9 (TLR9) in diabetic rats

2021 ◽  
Author(s):  
Simin Cai ◽  
Jing Li ◽  
Xiaoming Zhang
Keyword(s):  
Gut Microbiota ◽  
Kupffer Cells ◽  
Inflammatory Cytokine ◽  
Diabetic Rats ◽  
Toll Like Receptor ◽  
High Concentration ◽  
Cpg Dna ◽  
Significant Difference ◽  
Unmethylated Cytosine

Abstract Recently, gut microbiota for various pathogens has attracted attention. The present study investigated the role of gut microbiota unmethylated cytosine phosphate guanine DNA (CpG-DNA) on liver Kupffer cells (KCs) inflammatory cytokine interleukin-1β (IL-1β) in diabetic rats. We induced diabetic rats models and sequenced the gut microbiota composition of fecal samples. We also applied CpG-DNA and TLR9 inhibitor on KCs to investigate the regulation of inflammatory cytokine IL-1β and Toll-like receptor 9 (TLR9) signaling pathway. We found a significant difference of gut microbiota between the control and the diabetic rats with increased Clostridium. Meanwhile, diabetes could upregulate TLR9 in KCs and increase IL-1β concentration. Furthermore, high concentration of unmethylated CpG-DNA could significantly increase IL-1β secretion while it was suppressed by TLR9 inhibitor in KCs cultured in high glucose medium. Our study suggests that unmethylated CpG-DNA, which was highly expressed in diabetic rats, activated KCs through TLR9, and induced IL-1β secretion in vitro and in vivo which plays an important role in diabetic liver inflammation. It may contribute to the progress of the diabetes.

RES hyperphagocytosis by rats with streptozotocin-induced diabetes mellitus

1981 ◽  
Vol 240 (3) ◽  
pp. G225-G231
Author(s):  
R. P. Cornell
Keyword(s):  
Body Weight ◽  
Insulin Dose ◽  
Reticuloendothelial System ◽  
Diabetic Rats ◽  
Phagocytic Index ◽  
Control Value ◽  
Colloidal Carbon ◽  
Significant Difference

In contrast to previous studies of neutrophils from diabetic animals and humans in vitro and of macrophages from diabetic humans in vivo, which reported phagocytic depression, reticuloendothelial system (RES) hyperphagocytosis of colloidal carbon was observed in rats at 14 and 28 days after diabetes induction with streptozotocin (STZ). Carbon clearance half times were significantly enhanced to 6.3 +/- 0.79 and 8.1 +/- 1.04 min at 14 and 28 days post-STZ, respectively, compared with the nondiabetic value (12.7 +/- 0.98 min). The severity of uncontrolled STZ-induced diabetes in rats was confirmed by significant hypoinsulinemia, hyperglucagonemia, hyperglycemia, and hyperlipidemia. Although body weights of STZ-diabetic animals declined progressively, liver weights as a percent of body weight increased above the control value at 14 and 28 days post-STZ. In fact, expression of carbon phagocytosis as the corrected phagocytic index, which accounts for changes in liver and spleen weights relative to body weight, eliminated the significant difference between STZ-diabetic and nondiabetic animals. Antibiotic treatment of diabetic rats failed to alter the hyperphagocytosis, implying that a chronic bacterial infection was not the cause of phagocytic stimulation. Daily insulin replacements, but not a single large insulin dose to 14-day post-STZ rats, reversed the enhanced phagocytosis of colloidal carbon.

Endothelial cells but not platelets are the major source of Toll-like receptor 4 in the arterial thrombosis and tissue factor expression in mice

2014 ◽  
Vol 307 (7) ◽  
pp. R901-R907 ◽  
Author(s):  
Meiping Ren ◽  
Rong Li ◽  
Mao Luo ◽  
Ni Chen ◽  
Xin Deng ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Tissue Factor ◽  
Arterial Thrombosis ◽  
Vascular Wall ◽  
Toll Like Receptor ◽  
Occlusion Time ◽  
Significant Difference ◽  
Expression And Activity

It is known that Toll-like receptor (TLR)-4 plays an important role in myocardial infarction and atherothrombosis. The role of TLR-4 in arterial thrombosis is undefined. Both TLR-4-deficient ( TLR-4−/−) and wild-type (WT) mice were subjected to FeCl3carotid artery injury, and the time required to form an occlusive thrombus was measured. The mean time to occlusion in TLR-4−/−mice was significantly greater than that in WT mice after injury (303 ± 32 vs. 165 ± 34 s, P < 0.05). Furthermore, when we used a WT or TLR-4−/−-derived platelet reinfusion in a platelet depletion/reinfusion procedure, there was no significant change in the occlusion time and tissue factor (TF) activity in injured arteries between WT mice and platelet-depleted WT mice. Similarly, no significant difference was observed between TLR-4−/−mice and platelet-depleted TLR-4−/−mice for the WT or TLR-4−/−-derived platelet reinfusion. However, TF expression and activity were significantly reduced in the vascular wall of TLR-4−/−mice compared with WT mice. In vivo, lipopolysaccharide accelerated the occlusion time in WT mice but not TLR-4−/−mice. In vitro, LPS-induced TF activity was reduced in endothelial cells of TLR-4−/−mice relative to WT mice. The data demonstrate that TLR-4 contributes to arterial thrombosis formation in vivo and causes increased TF expression and activity in vitro. The results further suggest that the stimulation is mainly derived by endothelial cells but is not due to platelet-derived TLR-4.

scholarly journals NFATC3 promotes IRF7 transcriptional activity in plasmacy­­toid dendritic cells

2016 ◽  
Vol 213 (11) ◽  
pp. 2383-2398 ◽  
Author(s):  
Musheng Bao ◽  
York Wang ◽  
Ying Liu ◽  
Peiqing Shi ◽  
Hongbo Lu ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Transcription Factor ◽  
Nuclear Translocation ◽  
Constitutive Expression ◽  
Toll Like Receptor ◽  
Cpg Dna ◽  
Specialized Function

Plasmacytoid dendritic cells (pDCs) rapidly produce large amounts of type 1 interferon (IFN) after Toll-like receptor 7 and 9 engagements. This specialized function of type 1 IFN production is directly linked to the constitutive expression of IRF7, the master transcription factor for type 1 IFN production. However, the IRF7 regulatory network in pDCs remains largely unknown. In this study, we identify that the transcription factor NFATC3 specifically binds to IRF7 and enhances IRF7-mediated IFN production. Furthermore, knockout of NFATC3 greatly reduced the CpG DNA–induced nuclear translocation of IRF7, which resulted in impaired type 1 IFN production in vitro and in vivo. In addition, we found that NFATC3 and IRF7 both bound to type 1 IFN promoters and that the NFAT binding site in IFN promoters was required for IRF7-mediated IFN expression. Collectively, our study shows that the transcription factor NFATC3 binds to IRF7 and functions synergistically to enhance IRF7-mediated IFN expression in pDCs.

scholarly journals Bioconversion Variation of Ginsenoside CK Mediated by Human Gut Microbiota From Healthy Volunteers and Colorectal Cancer Patients

2021 ◽  
Author(s):  
Yin-Ping Guo ◽  
Li Shao ◽  
Li Wang ◽  
Man-Yun Chen ◽  
Wei Zhang ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
16S Rrna ◽  
Gut Microbiota ◽  
Healthy Volunteers ◽  
Healthy Subjects ◽  
Positive Association ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Significant Difference

Abstract Background: Ginsenoside CK (GCK) serves as the potential anti-colorectal cancer (CRC) protopanaxadiol (PPD)-type saponin, which could be mainly bio-converted to yield PPD by gut microbiota. Meanwhile, the anti-CRC effects of GCK could be altered by gut microbiota due to its different diversity in CRC patients. We aimed to investigate the bioconversion variation of GCK mediated by gut microbiota from CRC patients by comparing with healthy subjects.Methods: Gut microbiota profiled by 16S rRNA gene sequencing was collected from healthy volunteers and CRC patients. GCK was incubated with gut microbiota in vitro. A LC-MS/MS method was validated to quantify GCK and PPD after incubation at different time points.Results: The bioconversion of GCK in healthy subjects group was much faster than CRC group, as well as the yield of PPD. Moreover, significant difference of PPD concentration between healthy subjects group and CRC group could be observed at 12 h, 48 h and 72 h check points. According to 16S rRNA sequencing, the profiles of gut microbiota derived from healthy volunteers and CRC patients significantly varied, in which 12 differentially abundant taxon were found, such as Bifidobacterium, Roseburia, Bacteroides and Collinsella. Spearman’s correlation analysis showed bacteria enriched in healthy subjects group were positively associated with biotransformation of GCK, while bacteria enriched in CRC group displayed non correlation characters. Among them, Roseburia which could secrete β-glycosidase showed the strongest positive association with the bioconversion of GCK.Conclusion: The bioconversion of GCK in healthy subjects was much faster than CRC patients mediated by gut microbiota, which might alter the anti-CRC effects of GCK.

scholarly journals Eugenyl-2-Hydroxypropyl Methacrylate-Incorporated Experimental Dental Composite: Degree of Polymerization and In Vitro Cytotoxicity Evaluation

Polymers ◽  
2022 ◽  
Vol 14 (2) ◽  
pp. 277
Author(s):  
Abdel-Basit Al-Odayni ◽  
Dalal H. Alotaibi ◽  
Waseem Sharaf Saeed ◽  
Abdullah Al-Kahtani ◽  
Ali Assiri ◽  
...  
Keyword(s):  
Cell Viability ◽  
Colorimetric Assay ◽  
In Vitro Cytotoxicity ◽  
Degree Of Polymerization ◽  
Dental Composite ◽  
High Concentration ◽  
Dental Resins ◽  
Hydroxypropyl Methacrylate ◽  
Significant Difference

The aim of this study was to evaluate the properties of new dental formulations containing eugenyl-2-hydroxypropyl methacrylate (EgGMA) monomer, as restorative dental material, in terms of their degree of photopolymerization and cytotoxicity. The target model composites (TBEg0, TBEg2.5, and TBEg5) were prepared by mixing 35% organic matrix (TEGDMA/BisGMA (50/50 wt%) of which 0, 2.5, and 5 wt%, respectively, were replaced with EgGMA monomer) with 65% filler (silanized hydroxyapatite (HA)/zinc oxide (ZnO2), 4:3 by weight). The vinylic double-bond conversion (DC) after light-curing was studied using Fourier transform infrared technique whereas cell viability was in vitro tested using primary human gingival fibroblasts cells over 7 days by means of AlamarBlue colorimetric assay. The obtained data were statistically analyzed using ANOVA and Tukey post-hoc tests. The results revealed no significant difference in DC between TBEg2.5 (66.49%) and control (TBEg0; 68.74%), whereas both differ significantly with TBEg5, likely due to the inhibitory effect of eugenol moiety at high concentration. The cell viability test indicated that all the composites are biocompatible. No significant difference was counted between TBEg2.5 and TBEg5, however, both differed significantly from the control (TBEg0). Thus, even though its apparent negative effect on polymerization, EgGMA is potentially safer than bisphenol-derived monomers. Such potential properties may encourage further investigations on term of EgGMA amount optimization, compatibility with other dental resins, and antimicrobial activity.

Hyperbaric oxygenation affects acetylcholine-induced relaxation in female diabetic rats

2019 ◽  
pp. 635-646
Author(s):  
Dragan Manojlović ◽  
◽  
Ana Stupin ◽  
Zrinka Mihaljević ◽  
Anita Matić ◽  
...  
Keyword(s):  
Hyperbaric Oxygenation ◽  
Thiobarbituric Acid ◽  
Diabetic Rats ◽  
Sprague Dawley ◽  
Systemic Oxidative Stress ◽  
Reducing Ability ◽  
Significant Difference ◽  
And Control ◽  
Cox 1

We aimed to assess the effects of intermittent hyperbaric oxygenation (HBO2 at 2 bars for 120 minutes a day for four successive days) on acetylcholine-induced vasorelaxation (AChIR) in female Sprague-Dawley (SD) rats (N=80) that were randomized into four groups: healthy controls (CTR); diabetic rats (DM); and control and diabetic rats that underwent hyperbaric oxygenation (CTR+HBO and DM+HBO), respectively. AChIR was measured in vitro in aortic rings, with/without L-NAME, MS-PPOH, HET0016 or indomethacin. mRNA expression of eNOS, iNOS, COX-1, COX-2, thromboxane A synthase 1 (TBXAS1), CYP4A1, CYP4A3 and CYP2J3 was assessed by qPCR. Systemic oxidative stress and plasma antioxidative capacity were determined with the thiobarbituric acid-reactive substances (TBARS) and the ferric reducing ability of plasma (FRAP) assays, respectively. There was no significant difference in AChIR among experimental groups of rats. In CTR and DM group of rats, AChIR was mediated by NO and EETs pathway, while in the CTR+HBO and DM+HBO groups, NO-pathway prevailed. iNOS expression was upregulated in the DM group compared to CTR, while HBO2 upregulated eNOS in CTR group and TBXAS1 in DM group of rats. In both, CTR and DM group of rats, the sensitivity to ACh in the presence of L-NAME or in the presence of MSPPOH was significantly decreased compared to the response to ACh in the absence or presence of indomethacin or HET0016. DM and DM+HBO rats had increased TBARS compared to their respective controls. In conclusion, HBO2 presumably alters vasorelaxation in response to ACh from NO-EETs mediated pathways to solely NO-pathway, without affecting oxidative status of DM rats.

scholarly journals Expression of Phosphofructokinase-1 Gene in Streptozotocinized Diabetic Rats Fed Fermented and Non-Fermented Maize Diets

2021 ◽  
pp. 149-164
Author(s):  
Chukwudi Nze ◽  
Osaretin Albert Taiwo Ebuehi
Keyword(s):  
Gene Expression ◽  
Lipid Profile ◽  
Blood Chemistry ◽  
Diabetic Control ◽  
Diabetic Rats ◽  
Antioxidant Assay ◽  
Gene Expression Assay ◽  
Significant Difference ◽  
The Impact

Aim: This nutrigenomic research study is to investigate the impact of fermented maize (FM) and non-fermented maize (N-FM) diets on the expression of phosphofructokinase-1 (PFK-1) gene in a diabetic state. Methodology: The rats were equally grouped into four for the subsequent two weeks after acclimatization; Group 1 contained streptozotocinized-diabetic rats fed with FM diet (DFM), Group 2 contained streptozotocinized-diabetic rats fed with N-FM diet (DNM), Group 3 contained the normal control rats fed with standard rodent chow (NCG) and Group 4 contained diabetic control rats fed with standard rodent chow (DCG). The total phenol, flavonoid and antioxidant capacity (in vitro) of the maize diets were analyzed. Results: Rats fed the N-FM diet had higher concentration of phenols (73.20±0.9 mg/100 g) and flavonoids (82.83±1.02 mg/100 g). The in vitro antioxidant assay showed a statistically significant difference between the FM and N-FM diets (p<0.05). After the two weeks period, animals were sacrificed and blood samples obtained for blood chemistry and lipid profile tests. The livers were harvested for antioxidant activity and gene expression assay. The antioxidant assay showed no statistically significant difference among all groups, as well as the blood chemistry and lipid profile. The gene expression assay carried out using two-step Real-time qPCR, showed that PFK-1 gene was more expressed in the DFM group when compared to the DNM and DCG groups. Conclusion: The FM diet enhanced the expression of PFK-1 gene in streptozotocinized-diabetic rats.

scholarly journals PENGARUH KONSENTRASI MADU TERHADAP PERUBAHAN WARNA GIGI PADA PROSES PEMUTIHAN GIGI SECARA IN VITRO

2015 ◽  
Vol 1 (2) ◽  
pp. 25
Author(s):  
Sigit Fitri Istanti ◽  
Endah Aryati E ◽  
Kusuma Arbianti
Keyword(s):  
Negative Impact ◽  
T Test ◽  
Test Results ◽  
High Concentration ◽  
Tooth Discoloration ◽  
Significant Difference ◽  
Whitening Process ◽  
Colour Changes ◽  
Tooth Colour

Background: Needs of dental cosmetic services increased due to people's desire to get a brighter and whiter smile. Hydrogen Peroxide (H2O2) is one of bleaching substance that most commonly used. Use of H2O2 in a high concentration can cause a negative impact on the oral cavity. Honey is a natural ingredient that can whiten teeth. This study aimed to determine the effect of concentration of honey on tooth colour changes of teeth whitening process. Method: This research was an experimental in vitro study using pretest-posttest design with teeth as a research sample. 24 premolars teeth as samples were soaked in various concentration of honey (25%, 35%, 50%, 75%). The colour of teeth were measured with a spectrophotometer before and after treatment. Teeth colour were analyzed by paired samples T-test. Result: Paired T-test showed p<0.05, which means that concentration of honey affected on tooth colour changes. One way ANOVA test results showed concentrations of honey significantly affect tooth discoloration (p <0,05). Post hoc test results showed significant difference On the concentration of 25% of the concentration of honey 50%, 75%, but on a concentration of 25% honey and 35% there is no significant difference. Conclusion: From these results, it can be concluded that the concentration of honey affected on tooth colour changes in an in vitro tooth whitening process.

scholarly journals 83 CRYOPRESERVATION OF IN VITRO PORCINE OOCYTES BY SOLID SURFACE VITRIFICATION (SSV)

2005 ◽  
Vol 17 (2) ◽  
pp. 191 ◽  
Author(s):  
A. Dinnyes ◽  
T. Somfai ◽  
D. Sage ◽  
M. Marosan ◽  
J. Carnwath ◽  
...  
Keyword(s):  
Solid Surface ◽  
Electric Pulse ◽  
High Concentration ◽  
Parthenogenetic Activation ◽  
Surface Warming ◽  
National Office ◽  
Significant Difference ◽  
Pre Treatment ◽  
And Control

The effects of the cytosceletal inhibitor cytochalasin in solid surface vitrification (SSV; Dinnyes et al. 2000 Biol. Reprod. 63, 513–518) were investigated on in vitro matured (IVM) porcine oocytes. Cumulus-free IVM oocytes were subjected to one of the following: SSV (equilibration in 4% ethylene glycol (EG) for 10 min followed by vitrification in 35% EG, 5% polyvinyl pirrolidone, and 0.3 M trehalose on a cold (about −150°C) surface; warming in 0.4 M trehalose at 37°C), SSV pre-treatment with 5 μg/mL cytochalasin B (SSV + CB), or the steps of SSV without cooling, i.e. toxicity control (TC). Non-lysed oocytes together with the non-treated controls were subjected to parthenogenetic activation and then in vitro cultured (IVC) for six days. The proportion of non-lysed oocytes was higher when pre-treatment with CB was performed compared to SSV. However, both results were significantly lower than that of the TC. After parthenogenetic activation via a combination of a direct current electric pulse (1 kV/cm for 45 μs) followed by 3 h treatment with 2 mM 6-dimethylaminopurine, the proportion of cleaved embryos was higher in the SSV + CB than in the SSV. Nevertheless, significantly more oocytes cleaved in the TC and control groups. On Day 6 no blastocyst, were determined in the SSV group, and one blastocyst was obtained in the SSV + CB, while significantly more blastocysts developed in both the TC and the control (Table 1). There was no significant difference in blastocyst rates and in the number of blastomere nuclei/embryo between the TC and the control. These results indicate that the high concentration of cryoprotectants per se applied in SSV were not detrimental for in vitro development and that CB pre-treatment increased survival and further development of SSV vitrified pig oocytes resulting in one parthenogenetic blastocyst from vitrified pig oocytes. Table 1. Parthenogenetic development of vitrified IVM porcine oocytes This research was supported by a Bilateral Scientific and Technological Collaboration (TET, No. D6/01) grant between Germany and Hungary and by the Hungarian National Office of Research and Technology (OM-KMUFA; BIO-00086/2002).

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