Biochemical Liver Functions and Molecular Identification of Fasciola hepatica from Experimentally Infected Rat Model

The current study was performed to evaluate the liver function status as well as molecular characterization of the recovered worms in rats experimentally infected with F. hepatica. Sixteen male Wister rats aged 30 days were randomly allocated into two groups (n = 8). The first group was infected orally with 15 viable encysted metacercaria of F. hepatica per animal. The other group was kept non-infected (control group). At zero time (before infection), the 2nd, 4th, 6th, 8th, 10th, 12th and 14th weeks post-infection (WPI), blood and serum samples were collected via puncture of retro-orbital plexus of veins from each rat. Serum enzyme level (AST and ALT) and total protein were measured, and the serum protein profile was carried out using agarose gel electrophoresis. During the period of the experiment, serum ALT and AST activities and serum total globulins significantly increased while serum total proteins and albumin markedly decreased in the infected group. On the 14th WPI, the data of the electropherogram showed that globulin fractions (α1-, β- and γ-globulin) levels were significantly increased while α2-globulin was markedly decreased in the infected group. The molecular analysis confirmed the amplification of the ITS1, ITS2 and NDI genes of F. hepatica recovered from the infected liver of rats with amplicon sizes of 630, 510 and 560 bp, respectively. Sequencing of the amplified ITS gene resulted in the determination of 3 strains (PP108836, PP108837, and PP108838). Also, analysis of the ITS2 gene resulted in obtaining 3 isolates under the accession numbers (PP109065, PP109066, and PP109067). In conclusion, fasciolosis in the rat model is suitable for routine experimental infections and caused a pronounced liver dysfunction with discharging of the Fasciola eggs in the faeces and the development of adult stages in the bile ducts. Furthermore, molecular techniques are a sensitive tool for the identification and characterisation of the Fasciola parasite.

Phytochemical, antioxidant and hepatoprotective effects of different fractions of Moringa oleifera leaves methanol extract against liver injury in animal model

Objective: To evaluate the potential antioxidant and hepatoprotective effects of n-hexane, dichloromethane(DCM), ethyl acetate(EtOAc), n-butanol and aqueous fractions of Moringa oleifera(M. oleifera) leaves methanol extract against carbon tetrachloride(CCl_4)-induced liver injury in rats. Methods: These fractions were prepared from the M. oleifera leaves methanol extract by solubilization in water and partitioning in n-hexane, EtOAc, DCM and n-butanol. Their phyto-components were identified by GC-MS analysis. The in vitro antioxidant effect of these fractions was carried out by assessment of 1,1-diphenyl-2-picrylhydrazyl scavenging activity. A total of 40 Sprague Dawley rats were allocated into 8 equal groups: group 1 given olive oil(1 m L/kg b.wt.), group 2 injected with CCl_4, group 3 to 7 administered with n-hexane, DCM, EtOAc, n-butanol and aqueous fractions, respectively after CCl_4, group 8 administered with silymarin after CCl_4. The activities of aspartate aminotransferase, alanine aminotransferase, and the levels of total cholesterol, triglycerides, glucose, total proteins and albumin in serum were determined spectrophotometrically. Glutathione reduced, lipid peroxide by-products levels, glutathione-s-transferase and catalase enzyme activities in the liver homogenate were determined by spectrophotometer. Liver specimens were also examined for histopathological alterations under light microscope. Results: The GCMS analysis of different fractions of the M. oleifera leaves methanol extract revealed that n-hexane, DCM, EtOAc, n-butanol, and aqueous fractions contained 17, 22, 23, 19 and 32 compounds, respectively. The percent and the molecular structure of each component in each fraction were identified. The n-butanol and EtOAc fractions exhibited the strongest in vitro antioxidant activity against 1,1-diphenyl-2-picrylhydrazyl. CCl_4 significantly decreased glutathione reduced and total proteins concentration and glutathione-s-transferase and catalase activities but increased lipid peroxide by-products and total cholesterol levels. The n-hexane followed by aqueous and DCM fractions were the most potent to regulate serum enzyme activities and lipid peroxide by-products levels in the liver homogenate. Conclusions: n-hexane, DCM, and aqueous fractions have the highest effectiveness against CCl_4-induced hepatotoxicity. Isolation and purification of the active constituents require further experiments.

Phytochemical studies and anti-ulcerative colitis effect of Moringa oleifera seeds and Egyptian propolis methanol extracts in a rat model

Objective: To analyze the phytochemical constituents, and to explore potential protective effect of the methanol extract of Moringa oleifera(M. oleifera) seeds and Egyptian propolis, each alone or concurrently administered on acetic acid-induced ulcerative colitis in rats. Methods: Eight groups of 5 rats each were used: normal control group with distilled water, model group, two groups with M. oleifera seeds(100 and 200 mg/kg), two groups with propolis(50 and 100 mg/kg), one group with concurrent administration of both, and one group with prednisolone(reference drug). Macro-and microscopic picture, ulcer index and lesion scores, oxidative markers, inflammatory mediators, in vitro activity of the inflammatory enzymes and 1, 1-diphenyl-2-picrylhydrazyl free radicals scavenging activity were evaluated. The phytochemical constituents of both extracts were explored by GC-MS analysis. Results: Both treatments modulated the macro-and microscopic picture, decreased the ulcerative index, lesion score, oxidative markers and inflammatory mediators, and inhibited the COX-1 and COX-2 enzymes. Propolis appeared to be powerful free radicals scavenger. A powerful synergistic effect of both treatments in modulating the course of the disease was reported. GCMS analysis of methanol extract of M. oleifera seeds and propolis revealed the presence of 50 and 34 compounds, respectively. Conclusions: M. oleifera seeds and propolis methanol extracts have modulated the course of acetic acid-induced ulcerative colitis. Moreover, both treatments induce a good synergistic effect against the disease. Isolation of the active constituents is recommended.