Methylprednisolone induces activation of the contact system in a dose-dependent manner. An in vitro study

Abstract Background Cornin is a commonly used herb in cardiology for its cardioprotective effect. The effect of herbs on the activity of cytochrome P450 enzymes (CYP450s) can induce adverse drug-drug interaction even treatment failure. Therefore, it is necessary to investigate the effect of cornin on the activity of CYP450s, which can provide more guidance for the clinical application of cornin. Methods Cornin (100 μM) was incubated with eight isoforms of CYP450s, including CYP1A2, 2A6, 3A4, 2C8, 2C9, 2C19, 2D6, and 2E1, in pooled human liver microsomes. The inhibition model and corresponding parameters were also investigated. Results Cornin exerted significant inhibitory effect on the activity of CYP3A4, 2C9, and 2E1 in a dose-dependent manner with the IC50 values of 9.20, 22.91, and 14.28 μM, respectively (p < 0.05). Cornin inhibited the activity of CYP3A4 non-competitively with the Ki value of 4.69 μM, while the inhibition of CYP2C9 and 2E1 by cornin was competitive with the Ki value of 11.31 and 6.54 μM, respectively. Additionally, the inhibition of CYP3A4 by cornin was found to be time-dependent with the KI/Kinact value of 6.40/0.055 min− 1·μM− 1. Conclusions The inhibitory effect of cornin on the activity of CYP3A4, 2C9, and 2E1 indicated the potential drug-drug interaction between cornin and drugs metabolized by these CYP450s, which needs further investigation and validation.

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Hydroxyl Safflower Yellow Pigment A (HSYA) Improves Vascular Endothelial Injury Induced by Lipopolysaccharide In Vitro Study

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2021.2596 ◽

2021 ◽

Vol 11 (9) ◽

pp. 1792-1798

Author(s):

Li Yan ◽

Ge Jingping ◽

Yin Yuanyuan ◽

Li Xiaomei ◽

Zhao Boxiang ◽

...

Keyword(s):

Cell Proliferation ◽

In Vitro Study ◽

Treated Group ◽

Yellow Pigment ◽

Endothelial Injury ◽

Vitro Study ◽

Vascular Endothelial ◽

Treatment Groups ◽

Dose Dependent

Aim: This research was to investigate the effects and mechanisms of HSYA in vascular endothelial injury by vitro study. Methods: Dividing HUVECs as Normal Control (NC), Model (LPS treated) group, HSYA-L, HSYA-M and HSYA-H groups. Cells in the HSYA treatment groups were treated with LPS, followed by 40 mg/ml, 80 mg/ml, and 120 mg/ml HSYA intervention (HSYA-L, HSYA-M, and HSYA -H groups), respectively. Measuring the cell proliferation, apoptosis, relative proteins and mRNA (TLR4, MyD88 and NF-κB(p65)) expressions by MTT, Flow cytometry, WB and RT-qPCR assay. Using cellular immunofluorescence to evaluate NF-κB(p65) nuclear volume of difference groups. Results: With HSYA supplement, the cell proliferation rates were significantly up-regulation with cell apoptosis significantly down-regulation with TLR4 relatived mRNA and proteins and NF-κB(p65) nuclear significantly depressed with dose-dependent (P <0.05, respectively). Conclusion: HSYA improved vascular endothelial injury induced by LPS via TLR4 pathway In Vitro.

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Ascorbic acid attenuates activation and cytokine production in sepsis-like monocytes

10.1101/2021.04.15.21255504 ◽

2021 ◽

Author(s):

Tobias Schmidt ◽

Robin Kahn ◽

Fredrik Kahn

Keyword(s):

Flow Cytometry ◽

Ascorbic Acid ◽

Cytokine Production ◽

In Vitro Study ◽

Surface Expression ◽

High Dose ◽

Functional Assay ◽

Dependent Manner ◽

Dose Dependent

Objective To investigate the effects of high dose ascorbic acid (AA) on monocyte polarization and cytokine production in vitro Design Experimental in vitro study of cells from healthy subjects and patients with sepsis Setting University research laboratory and academic hospital Subjects Six healthy controls and three patients with sepsis Interventions Monocytes were isolated from whole blood of healthy donors (n=6) and polarized in vitro for 48hrs using LPS or LTA. Polarization was confirmed by surface marker expression using flow cytometry. As a comparison, monocytes were also isolated from septic patients (n=3) and analyzed for polarization markers. The effect of AA on monocyte polarization was evaluated. As a functional assay, AA-treated monocytes were analyzed for cytokine production of TNF and IL-8 by intracellular staining and flow cytometry following activation with LPS or LTA. Measurements and Main Results Both LPS and LTA induced polarization in healthy monocytes in vitro, with increased expression of both pro- (CD40 and PDL1, p<0.05) and anti-inflammatory (CD16 and CD163, p<0.05) polarization markers, with non-significant effects on CD86 and CD206. This pattern resembled, at least partly, that of monocytes from septic patients. Treatment with AA significantly inhibited the upregulation of surface expression of CD16 and CD163 (p<0.05) in a dose dependent manner, but not CD40 or PDL-1. Finally, AA attenuated LPS or LTA-induced cytokine production of IL-8 and TNF in a dose-dependent manner (both p<0.05). Conclusions AA inhibits upregulation of anti-, but not pro-inflammatory related markers in LPS or LTA polarized monocytes. Additionally, AA attenuates cytokine production from in vitro polarized monocytes, displaying functional involvement. This study provides important insight into the immunological effects of high dose AA on monocytes, and potential implications in sepsis.

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INVESTIGATION ON PHARMACOGNOSY AS WELL AS THE ANTIOXIDANT, ANTI-INFLAMMATORY POTENTIAL OF THE KATHA POWDER

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2021.v14i6.41710 ◽

2021 ◽

pp. 125-132

Author(s):

PANKAJ SHARMA ◽

RAJU L

Keyword(s):

Diclofenac Sodium ◽

In Vitro Study ◽

Alcoholic Solution ◽

Hot Water ◽

Dependent Manner ◽

Standard Drug ◽

Anti Inflammatory ◽

Acacia Catechu ◽

Dose Dependent

Objective: The objective of the study was to investigate the pharmacognosy as well as the antioxidant, anti-inflammatory potential of the Katha powder. Methods: The Coarsely dried chips of Acacia catechu heartwood were treated with 10 % hydro-alcoholic solution to obtain Katha as the final product. The powdered Katha was standardized through pharmacognostic parameters. This Katha power is showing the good solubility in the hot water having astringent in the taste. The powder microscopy of the Katha powder is to be demonstrated fragments of acicular crystals, fibers, and bordered pitted vessels. Katha powder antioxidant potential is to be accessed by using the 2, 2-diphenyl-1-picryl hydrazyl assay and NO Scavenging assay using ascorbic acid as a standard drug. Further, the Katha powder is to be subjected for the assessment of its anti-inflammatory potential by the use of heat-induced hemolysis as well as hypotonicity-induced hemolysis approach by the use of the aspirin or diclofenac sodium as a standard drug. Results: Microscopical investigations were showed that Katha showing the presence of fragments of acicular crystals, fibers, and bordered pitted vessels. In vitro study shows that the Katha powder has excellent antioxidant as well as anti-inflammatory potential in a dose-dependent manner in comparison of the result of heartwood of A. catechu. Conclusion: So from this investigation, it is to be suggested that the Katha powder is rich in the phenolic compound and the experimentation study shows that the drug is to possess a good antioxidant as well as anti-inflammatory property.

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Obestatin Modulates Ghrelin’s Effects on the Basal and Stimulated Testosterone Secretion by the Testis of Rat: an In Vitro Study

Physiological Research ◽

10.33549/physiolres.933345 ◽

2017 ◽

pp. 93-98

Author(s):

T. AFSAR ◽

S. JAHAN ◽

S. RAZAK ◽

A. ALMAJWAL ◽

M. ABULMEATY ◽

...

Keyword(s):

In Vitro Study ◽

Inhibitory Effect ◽

Human Chorionic Gonadotrophin ◽

Dependent Manner ◽

Control Groups ◽

Adult Rats ◽

Testosterone Secretion ◽

Functional Antagonism ◽

Dose Dependent

The functional antagonism between obestatin and ghrelin in the testis is under investigation. We investigated the ability of obestatin to counteract the inhibitory effect of ghrelin on basal and stimulated testosterone (T) secretion in vitro. Testicular strips from adult rats were incubated with 10 ng/ml and 100 ng/ml of obestatin alone, ghrelin alone and obestatin + ghrelin. Obestatin modulation of stimulated T secretion was evaluated by incubation of testicular samples with 10 ng/ml and 100 ng/ml obestatin, ghrelin and obestatin + ghrelin in the absence and presence of 10 IU of human chorionic gonadotrophin (hCG). T concentrations in the hCG treated groups were significantly (P<0.0001) higher than those in the control groups. Obestatin caused a significant increase in basal T secretion in a dose-dependent manner; however, obestatin at the both 10 ng/ml and 100 ng/ml significantly (P<0.0001) increased hCG-stimulated T secretion. In contrast, ghrelin in a dose-dependent manner significantly (P<0.001) decreased both basal and hCG-induced T secretion by testicular slices. Obestatin opposed the inhibitory effect of ghrelin on T secretion under both basal and hCG-stimulated conditions at all doses tested. In conclusions, administration of obestatin was able to antagonize the inhibitory effect of ghrelin on testosterone secretion in vitro.

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TAMARIND EXTRACT INHIBITS CYTOCHROME P450 (CYP3A4 ISOZYME) - AN IN VITRO STUDY

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2018.v11i7.25767 ◽

2018 ◽

Vol 11 (7) ◽

pp. 333

Author(s):

Jagadish Rajkumaar R ◽

Anitha Roy ◽

Lakshmi T

Keyword(s):

Room Temperature ◽

Potassium Phosphate ◽

In Vitro Study ◽

Inhibitory Effect ◽

Dependent Manner ◽

Fruit Pulp ◽

Ic50 Value ◽

Dose Dependent ◽

Cytochrome P 450

Objective: The aim of the present study was to analyze the effect of the aqueous fruit pulp extract of Tamarindus indica L. (tamarind extract) on cytochrome P 450 isoform CYP3A4.Methods: Tamarind extract at different concentrations from 5 to 100 μg/ml was examined for its inhibitory property toward cytochrome P 450 isoform CYP3A4. The various concentrations of tamarind extract, potassium phosphate buffer, CYP450 reagent, and substrate 7-Benzyloxy-4- trifluoromethylcoumarin were added to a 96-well plate. The mixtures were preincubated for 20 min at room temperature. The reaction was started by a mixture of free constituted substrate and NADP+ and incubated at room temperature for 30–60 min. The reaction was stopped by Tris-HCl buffer, pH 10.5. The fluorescent intensities of the products were measured by PerkinElmer Enspire fluorescence reader using an excitation and emission wavelength of 405 nm and 460 nm, respectively. Inhibitory concentration (IC50) was calculated by plotting concentrations of tamarind extract against the corresponding percentage inhibition.Results: All the tested concentrations of extract except 5 μg/ml showed good inhibition against CYP3A4 in a dose-dependent manner. The IC50 value of tamarind for CYP3A4 inhibitory activity was found to be 27.89 μg/ml.Conclusion: T. indica aqueous fruit pulp extract exhibited an inhibitory effect on CYP34A, thereby indicating the possibilities of herb-drug interaction if these extracts are coadministered with the prescribed drugs that are metabolized by CYP3A4.

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Itraconazole and fluconazole-induced toxicity in rat hepatocytes: a comparativein vitro study

Human & Experimental Toxicology ◽

10.1191/0960327102ht208oa ◽

2002 ◽

Vol 21 (1) ◽

pp. 43-48 ◽

Cited By ~ 12

Author(s):

N Somchit ◽

S M Hassim ◽

S H Samsudin

Keyword(s):

Cytochrome P450 ◽

Lactate Dehydrogenase ◽

Rat Hepatocytes ◽

Antifungal Drugs ◽

Vitro Study ◽

Time Dependent ◽

In Vitro Toxicity ◽

Dependent Manner ◽

Dose Dependent

This current study was to investigate the in vitrocytotoxicity of rat hepatocytes induced by the antifungal drugs, itraconazole and fluconazole. Both antifungal drugs caused dose-dependent cytotoxicity. In vitro incubation of hepatocytes with itraconazole revealed significantly higher lactate dehydrogenase (LDH) leakage when compared to fluconazole. Phenobarbital pretreated hepatocytes contained significantly higher total cytochrome P450 content than the control hepatocytes. P450 content was reduced approximately 30% for both types of hepatocytes after 6 hours incubation. Interestingly, cytotoxicity of itraconazole was reduced significantly by phenobarbital pretreatment. Phenobarbital did not have any effect on the cytotoxicity induced by fluconazole. These results demonstrate the in vitro toxicity of hepatocytes induced by itraconazole and fluconazole that were expressed in a dose and time-dependent manner. Phenobarbital plays a role in the cytoprotection of hepatocytes to itraconazole-induced but not fluconazole-induced cytotoxicity in vitro.

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