Open Access Original Research Article

LIGHT and FGF-BP are Upregulated in Chronic Liver Injury

Elsayed Gomaa Elsayed Elsakka

International Journal of Biochemistry Research & Review, Page 1-9
DOI: 10.9734/IJBCRR/2016/26029

Aims: The aim of this study is to explore the expression pattern of LIGHT and FGF-BP at the different stages of liver injury including acute injury, fibrosis and cirrhosis.

Study Design: Controlled experiment. 

Place and Duration of Study: Department of Biochemistry and Department of Pharmacology and Toxicology, Faculty of pharmacy (boys) Al-Azhar University, between February 2015 and June 2015.

Methodology: Four Sprague-Dawley rat groups were used for the experiment. Control group: 9 rats received corn oil; acute toxicity group: 10 rats were injected 50% CCl4 in corn oil (4 ml/kg/IP/single dose); 6 weeks group: 12 rats were injected with 50% CCl4 in corn oil (4 ml/kg/IP/twice weekly/6 weeks); 11 weeks group: 10 rats were injected with 50% CCl4 in corn oil (4 ml/kg/IP/twice weekly/6 weeks) followed by 5 weeks of CCl4 treatment with the half previous dose.

On the day after the last dose, rats were anesthetized with diethyl ether and blood samples were collected for measurement of blood chemistry. The animals were then euthanized, and tissue samples from the livers harvested and divided into 2 parts; one part was processed for standard histology and immunofluorescence techniques and the other was homogenized for oxidative status assessment.

Results: LIGHT and FGF-BP were shown to upregulate in chronic liver injury; while LIGHT was shown to upregulate in cirrhosis stage only, FGF-BP was shown to upregulate at fibrosis and cirrhosis stages. LIGHT was shown to express at the cytoplasm of the cells at cirrhotic nodule border. FGF-BP localization was shown to occur mainly at the extracellular matrix.

Conclusion: LIGHT and FGF-BP are of critical importance in pathogenesis of chronic liver injury so they may be used as a target for chronic liver injury therapy and/or candidate marker for diagnosis and/or prognosis of chronic liver injury.

Open Access Original Research Article

Characterization of Elastolytic Protease, Asnilase from Aspergillus nidulans and Its Cytotoxicity on Human Endothelial and Epithelial Cells

Yumiko Komori, Yoshiyuki Okumura, Kazuhito Kamiya, Kenji Ogawa, Toshiaki Nikai

International Journal of Biochemistry Research & Review, Page 1-8
DOI: 10.9734/IJBCRR/2016/26206

Aims: To elucidate the pathogenicity of Aspergillus nidulans, elastolytic protease was isolated from the culture supernatant of the isolate, and its biological activity and cytotoxicity were examined.

Methodology: A. nidulans (NBRC 4340) spores were cultured on YCB-elastin medium for three days and elastolytic protease was isolated from the culture supernatant by DEAE-Cellulose anion exchange chromatography. Elastolytic activity was measured by using GAAPLNA (Glutaryl-L-alanyl-L-alanyl-L-prolyl-L-leucine p-nitroanilide) as the substrate. Molecular mass and isoelectric point were determined by polyacrylamide gel electrophoresis. Substrate specificities were measured by using fibrinogen, collagen, and oxidized insulin B chain. Cytotoxicity of Asnilase on human pulmonary artery endothelial cells (HPAEC), human small airway epithelial cells (HSAEC), human bronchial/tracheal epithelial cells (HBTEC) and human pulmonary alveolar epithelial cells (HPAEpiC) was determined with colorimetric method.

Results: An elastolytic protease isolated from the culture supernatant of A. nidulans was found to be homogeneous as indicated by a single band after polyacrylamide gel electrophoresis (PAGE), and the final preparation named as Asnilase. Molecular mass of this enzyme was determined to be 33,800 Da and the isoelectric point was 4.2. Asnilase cleaved the Aα and Bβ chains of fibrinogen, collagen and hydrolyzed His(5)-Leu(6) and Glu(13)-Ala(14) bonds of oxidized insulin B chain. Cytotoxic effects for various human pulmonary cells were observed when Asnilase was added at concentration of 2.0-8.0 mg/mL.

Conclusion: It is shown in our current investigation that Asnilase is a newly isolated elastolytic protease from A. nidulans. It is responsible for pulmonary aspergillus infection.

Open Access Original Research Article

Chelation Potential of Aqueous Leaf Extracts of Vernonia amygdalina and Phyllanthus amarus on Kidney Functions in Lead-intoxicated Albino Wistar Rats

U. Ezirim Amanda, C. Udenze Emeka, P. Ihedimbu Chiamaka, I. Ukairo Doris, I. Iheme Callistus

International Journal of Biochemistry Research & Review, Page 1-10
DOI: 10.9734/IJBCRR/2016/25356

The aim of the study was to investigate the chelation potential of aqueous leaf extracts of Vernonia amygdalina and Phyllanthus amarus in albino wistar rats intoxicated with lead using kidney function markers as indicators, and compare results to an EDTA chelation therapy. Both plants were assessed for phytochemicals. Forty-two male albino wistar rats with mean weight of 75 g were divided into seven groups of six animals each: normal NC (normal saline) and intoxicated control IC (1000 ppm lead acetate in water) for three weeks; Drug treated control DTC (75 mg/kg Na2EDTA co-administered with 5 mg/kg Calcium); V. amygdalina-treated IT1 (100 mg/kg) and IT2 (200 mg/kg); P. amarus-treated IT3 (100 mg/kg) and IT4 (200 mg/kg) for eight consecutive days after three weeks of exposure. Biomarkers analyzed include serum concentrations of urea, creatinine, and electrolytes. Histological assessments on kidney tissues were performed. A significantly elevated (p < 0.05) trend was detected in serum urea (17.93±2.99 mmol/L) and creatinine (4.28±1.38 mg/dL) levels of the intoxicated control IC. The DTC (10.16±1.46 mmol/L), 100 mg/kg VA(12.69±1.59 mmol/L), 200 mg/kg VA(10.48±1.12 mmol/L), 100 mg/kg PA(9.96±1.83 mmol/L) and 200 mg/kg PA (9.12±1.19 mmol/L) treatment groups showed significant (p < 0.05) decrease in serum urea concentrations close to NC (10.55±1.20 mmol/L). Similar significant reversal (p < 0.05) of the elevated serum creatinine levels were observed in DTC (2.60±1.08 mg/dL), 100 mg/kg VA(2.22±0.43 mg/dL), 200 mg/kg VA(2.07±0.416 mg/dL), 100 mg/kg PA(1.99±0.27 mg/dL) and 200mg/kg PA (2.04±0.28 mg/dL). The intoxicated control IC exhibited remarkable decline (p < 0.05) in serum sodium (69.67±8.59 mmol/L) and potassium (4.93±0.71 mmol/L) concentrations. These were significantly elevated (p < 0.05) in all treatment groups. Histological examinations confirmed the amelioration of deranged tissues in the treatment groups. Aqueous leaf extracts of                             V. amygdalina and P. amarus can compete favorably with an EDTA chelation therapy, with regard to reversing impaired kidney functions as a result of lead intoxication.

Open Access Original Research Article

Ethanol Extract of Acalypha wilkesiana Muel Arg Leaves Ameliorates Paracetamol-induced Hepatotoxicity in Rats

C. L. Onuah, C. C. Monago, S. I. Omeodu

International Journal of Biochemistry Research & Review, Page 1-7
DOI: 10.9734/IJBCRR/2016/25588

Objective: The effect of ethanol leaf extract of A. wilkesiana on the liver of paracetamol- induced hepatotoxicity was investigated in wistar albino rats.

Study Design: Animal experimental study.

Place of Study: Department of Biochemistry, Faculty of Biological science University of Port Harcourt P.M.B 5323 Port Harcourt Nigeria

Methods: Liver toxicity was induced with 2000 mg/kg body weight of paracetamol (PARA) orally. The extract was administered to paracetamol treated wistar albino rats at a dose of 100 mg/kg,     200 mg/kg and 300 mg/kg body weight. Qualitative phytochemical analysis of the leaves of                            A. wilkesiana (AW) showed that they are rich in flavonoids, phenols, tannins, terpenoids, cardiac glycosides, protein, alkaloids and steroids. The effect of the ethanol leaf extract of A. wilkesiana on the liver was monitored by measuring the liver enzymes (alkaline phosphatase, alanine transaminase and aspartate transaminase), total bilirubin, albumin and total protein.

Results: The ethanol leaf extract of A. wilkesiana significantly (p<0.05) lowered the activities of alkaline phosphatase, alanine and aspartate transaminases.

Conclusion: Histopathological studies showed that ethanol leaf extract of A. wilkesiana had a therapeutic effect on the liver of wistar albino rats with paracetamol induced hepatotoxicity.

Open Access Review Article

Phytochemistry and Antimicrobial Activity of Extracts from Medicinal Plant Olea africana and Olea europea

Kemboi Douglas

International Journal of Biochemistry Research & Review, Page 1-7
DOI: 10.9734/IJBCRR/2016/25863

Increase in prevalence of resistant microorganisms especially to synthetic drugs, has necessitated the need to search for new bioactive compounds having natural origin. Thus it was essential to determine the antibacterial activity of hexane, dichloromethane, ethyl acetate and methanol extracts of Olea africana and Olea europea. Phytochemical investigation of both Olea africana and             O. europea extracts indicated the presence alkaloids, flavones, flavanoids and tannins. Olea africana leaves extracts afforded two triterpenoids namely erythrodiol and uvaol which were obtained through repeated column chromatography. The compounds were characterized using Nuclear Magnetic Resonance (NMR) spectroscopy, MS spectroscopy and by comparison with literature values. The isolated triterpenoids exhibited moderate antibacterial activity whereas crude extracts exhibited relatively high antibacterial activity against Gram positive bacterial strains; methanol extracts inhibited growth and showed 12.4 mm zone of inhibition against Staphylococcus aureus, Erythrodiol exhibited higher antibacterial activity than uvaol against Gram positive bacteria Staphylococcus aureus with zones of inhibition of 5.2 mm and 5.0 mm respectively. None of the pure compounds showed significant activity against Gram negative bacteria Escherichia coli.  The results gives a scientific validity and credence to the ethno-medicinal use of this medicinal plant as a chewing stick.