EXCLI Journal

Topical phenytoin accelerates the healing of acetic acid-induced colitis in Rats: evaluating of Transforming growth factor-beta, Platelet-derived growth factor, and Vascular endothelial growth factor.

2021-04-15T12:42:28+02:00

BACKGROUND: Ulcerative colitis (UC) limited to the innermost lining of the colon has become a global health problem. Immunomodulatory and monoclonal antibodies are using despite their side effects and limitations of effective choices. Phenytoin is using to heal wounds due to its side effects like the increased expression of growth factors, collagen, and extracellular matrix synthesis.

OBJECTIVE: This study aimed to evaluate the effect of topical phenytoin in UC.

DESIGN: Phenytoin in two-doses and period treatment had investigated.

SETTINGS: Male Wistar rats (230–280 g, n=8) were divided randomly into ten groups.

PATIENTS: This study didn't use the human-related protocol.

INTERVENTIONS: Animals divided as follows: Sham, control, hydrocortisone, phenytoin 1%, and 3% groups in six or 12-days treatment protocol. Ulcerative colitis model was induced by administration acetic acid 4% into the colon transracially animals were sacrificed at postoperative days 7 and 13.

MAIN OUTCOME MEASURES: Bodyweight loss, Microscopic score, and Ulcer index measured using specific criteria, and growth factors were measure by western blotting.

RESULTS: Bodyweight loss reversed in treatment groups. Ulcer index in six and 12-day had reduced. Microscopic score in six-day enema treatment significantly decreased in treatment groups compared to the control groups. TGF-β in a time-dependent and PDGF and VEGF time and dose-dependently in phenytoin 1% and3% significantly increased in six and 12-day protocol.

LIMITATIONS: Acetic acid-induced UC in rats may not be matched wholly with the development and process of UC in humans.

CONCLUSIONS: Phenytoin dose and time-dependently reversed weight loss. Also, histopathological parameters include microscopic scores, and ulcer index was decreased via the induction of growth factors TGF-β, PDGF, and VEGF and accelerate ulcer healing.

Lungs epithelial cells injury induced by C-reactive protein (CRP) is mediated via p38 MAPK/mitochondrial apoptosis pathway

2021-04-15T12:30:18+02:00

Increased level of C-reactive protein (CRP) are known to be associated with Mycoplasma pneumoniae (MP) infection. The purpose of this study was to explore whether C-reactive protein (CRP) causes the apoptosis of lung epithelial cells after MP infection and the underlying mechanism. M. pneumoniae pneumonia (MPP) patients were included for CRP detection using immuno-turbidimetry and western blot. The expression of CRP in A549 cells was detected by immuno-fluorescence, qRT-PCR, western blot and immunoturbidimetric. The viability of A549 cells was detected by MTT, the expressions of p38 MAPK/mitochondrial apoptotic pathway-related proteins were evaluated by western blot, and the apoptosis was determined by flow cytometry. The expressions of Cyt C and Tom 20 were observed by immuno-fluorescence, the mitochondrial membrane potential (ΔΨm) was detected by JC-1, and ROS content was measured by DCFH-DA. We found that the expression of CRP was elevated in blood of MPP patients. The expression of CRP in MP-infected A549 cells increased significantly. The cell viability was significantly reduced, the cell morphology was abnormal and the rate of apoptosis was markedly increased in MP group. Next, overexpression of CRP induced the up-regulation of the p38 MAPK/mitochondrial apoptosis pathway related protein expression levels and increased Cyt C expression in the cytoplasm, decreased Tom 20 expression and ΔΨm and up-regulated ROS expression. The addition of SB203580 and CsA alleviated the damage of CRP to A549 cells. The increased CRP induced by MP infection, caused apoptosis of lung epithelial cells through the p38 MAPK/mitochondrial apoptosis pathway.

In silico design and immunological evaluation of a novel recombinant epitope-based protein of MDR Acinetobacter baumannii

2021-04-15T00:23:06+02:00

Vaccination is a promising strategy for the life-threatening MDR A. baumannii. In this study, we directed to design and evaluate immunological efficacy of a recombinant multi-epitope protein (rMEP). Epitope prediction was performed for candidate proteins OmpA and BAM complex (BamA, BamB, BamC, BamD and BamE) of A. baumannii, using immune-informatics tools with high affinity for the human HLA alleles. The ELISA, Western blot and Opsonic killing assay were applied for immunogenic reaction of designed recombinant protein. Results showed significant interaction designed protein with ICU patients and staff serum antibodies. There was a significant correlation between total antibody titer against rMEP and opsonic killing activities (correlation coefficient 0.67 and P< 0.001). Circular Dichroism study of purified protein showed the secondary structure and refolding of recombinant protein properly done and was more-matched with in silico prediction. The interaction of infected patient’s serum antibodies with the recombinant protein and bactericidal activity of the same serums against the A. baumannii, suggest the rMEP is a promising immunogenic protein for protection of MDR A. baumannii. Further studies are needed for investigation of different aspect of rMEP including in vivo studies.