Rosmarinic acid attenuates hepatic fibrogenesis via suppression of hepatic stellate cell activation/proliferation and induction of apoptosis

OBJECTIVE@#To investigate the antifibrotic role of rosmarinic acid (RA), a natural polyphenolic compound, on HSCs activation/proliferation and apoptosis in vitro and in vivo.@*METHODS@#The impact of RA on stellate cell line (HSC-T6) proliferation, activation and apoptosis was assessed along with its safety on primary hepatocytes. In vivo, rats were divided into: (i) normal; (ii) thioacetamide (TAA)-intoxicated rats for 12 weeks; (iii) TAA + silymarin or (iv) TAA + RA. At the end of experiment, liver functions, oxidative stress, inflammatory and profibrogenic markers, tissue inhibitor metalloproteinases type-1 (TIMP-1) and hydroxyproline (HP) levels were evaluated. Additionally, liver histopathology and immunohistochemical examinations of alpha-smooth muscle actin (α-SMA), caspase-3 and proliferation cellular nuclear antigen (PCNA) were determined.@*RESULTS@#RA exhibited anti-proliferative effects on cultured HSCs in a time and concentration dependent manner showing an IC of 276 μg/mL and 171 μg/mL for 24 h and 48 h, respectively, with morphological reversion of activated stellate cell morphology to quiescent form. It significantly improved ALT, AST, oxidative stress markers and reduced TIMP-1, HP levels, inflammatory markers and fibrosis score (S1 vs S4). Furthermore, reduction in α-SMA plus elevation in caspase-3 expressions of HSCs in vitro and in vivo associated with an inhibition in proliferation of damaged hepatocytes were recorded.@*CONCLUSIONS@#RA impeded the progression of liver fibrosis through inhibition of HSCs activation/proliferation and induction of apoptosis with preservation of hepatic architecture.

Rosmarinic acid attenuates hepatic fibrogenesis via suppression of hepatic stellate cell activation/proliferation and induction of apoptosis

Objective To investigate the antifibrotic role of rosmarinic acid (RA), a natural polyphenolic compound, on HSCs activation/proliferation and apoptosis in vitro and in vivo. Methods The impact of RA on stellate cell line (HSC-T6) proliferation, activation and apoptosis was assessed along with its safety on primary hepatocytes. In vivo, rats were divided into: (i) normal; (ii) thioacetamide (TAA)-intoxicated rats for 12 weeks; (iii) TAA + silymarin or (iv) TAA + RA. At the end of experiment, liver functions, oxidative stress, inflammatory and profibrogenic markers, tissue inhibitor metalloproteinases type-1 (TIMP-1) and hydroxyproline (HP) levels were evaluated. Additionally, liver histopathology and immunohistochemical examinations of alpha-smooth muscle actin (α-SMA), caspase-3 and proliferation cellular nuclear antigen (PCNA) were determined. Results RA exhibited anti-proliferative effects on cultured HSCs in a time and concentration dependent manner showing an IC

use of microencapsulated hepatocytes transplantation reduces mortality and liver alterations in schistosoma mansoni infected hamsters

Hepatocyte transplantation is an attractive therapeutic modality for liver disease as an alternative for orthotropic liver transplantation. The goal of this work was to study the adequacy of intrasplenic hepatocyte transplantation [HCTx] in fresh and microencapsulated forms, in a hamster model of liver fibrosis by Schistosoma mansoni infected hamsters were divided into 6 groups; untreated for 11 weeks [GI] and for 15 weeks [GII] treated with praziquantel [PZQ] 7 weeks PI, and killed 4 weeks [GUI] and 8 weeks [GIV] post-treatment. Treated with PZQ 7 weeks PI, and then treated orally with immunosuppressive drug "cyclosporine [4 weeks post PZQ treatment], 24 hr. before interasplenic injection with fresh hepatocytes [V]. Treated with PZQ 7 weeks PI, and then injected interasplenically [4 weeks post-treatment] with microencapsulated hepatocytes [GVI]. GI and GUI were killed 11 weeks PI for assessment the anti-schistosomal efficacy of PZQ. The other four groups were killed 15 weeks PI for investigation of liver and spleen histology, serum liver enzymes and hepatic oxidative markers before and after HCTx. Freshly isolated hepatocytes with a mean viability 92.9711.2% were used for microencapsulation and transplantation. Histological study showed the presence of transplanted hepatocytes in spleen of recipient. PZQ accelerated healing of hepatic granulomatous lesions as evidenced parasitologically by the increase in the percentage of dead eggs and histologically showing more gfanuloma circumscription with more ova degeneration and less inflammatory cells. The 25-day survival rates in Gil, GIV, GVand GVI were 5/15 [33.3%], 8/15 [53.3%], 10/15 [66.7%] and 9/15 [60%] respecttively. In addition, there were significantly better outcomes in serum biochemical indexes such as ALT, AST, y-GT, ALP, and hepatic SOD and MDA in the fresh and microencapsulated groups than in PZQ-treated group, without great differences between the microencapsulated and the fresh transplanted groups. Liver pathological staining supported these findings

Pharmacological and antioxidant actions of garlic and/or onion in non-alcoholic fatty liver disease [NAFLD] in rats

Non-alcoholic fatty liver disease [NAFLD] includes a broad spectrum of fat-induced liver injury, ranging from mild steatosis to cirrhosis and liver failure. This study investigates the hepatoprotective properties of garlic and onion in NAFLD rat model. Ninety male Sprague-Dawley rats were randomly divided into 9 groups; normal [I], NAFLD induced with high fat diet [HFD; II], NAFLD switched to regular diet [RD; III], NAFLD-HFD or NAFLD-RD treated with garlic [IV, V] onion [VI, VII] or the combined garlic+onion [VIII, IX] respectively. A NAFLD rat model was established by feeding the animals with a high-fat diet for 12 wk. These animals were then treated with garlic or/and onion or vehicle for 8 wk [weeks 13-20] and then killed to obtain serum samples and liver tissues. Liver histology, lipids, parameters of oxidative stress, TNF-alpha and TGF-p were measured. The liver in NAFLD-HFD showed typical steatosis, accompanied with mild to moderate lobular inflammatory cell infiltration. Serum levels of ALT, AST, ALP, leptin, cholesterol, triglycerides, TNF-alpha, TGF-P and hepatic MDA were significantly increased [P<0.05] compared with normal group. This was accompanied with reduction of hepatic GSH, GR, GPx, GST, SOD and serum adiponectin. These changes were to a less degree in NAFLD-RD group. Combined administration of garlic+onion produced a better and significant decrease in liver steatosis, serum liver enzymes, oxidative markers and lipid peroxidation versus each one alone. In the same time, NAFLD-induced inflammation was also mitigated via reduction of TNF-alpha and TGF-P. In addition, these results were better in the group IX versus group VIII

Susceptibility of giardia lambia to newly introduced synthetic compounds in experimentally infected animals

Therapeutic strategy has included diverse pharmaceutical agents of traditional use such as metronidazole, quinacrine, furazolidone and paramomycin, other drugs of more recent introduction, such as albendazole and nitazoxanide, have also been applied as clinical practice. Of these e.g. metronidazole may be considered the most representative anti-giardial agents of traditional and recent use. However, evidence points to an increasing frequency of cases refractory to treatment with these drugs, the causes of which include non-compliance to treatment and emergence of drug resistant Giardia. The aim of this study is to evaluate the therapeutic effect of four new compounds, 3- / [[2E]-3-[dimethylamino] Prop-2-enoyl/ ] -5, 6-diphenyl- 1, 2, 4-triazin-3 [2H]-one [T1] 3-/ [[2E]-3-[dimethylamino] prop2-enoyl]-2H-chromen-2-one [C1] and 3-[Hexa-2, 4 dienoyl]- 4 hydroxy-1-methyl quinolin-2 [1H]-one [Q1] and N, N-Bis / [2,4-dioxo-1-methyl-1, 2, 3, 4-tetrhydroquinol-3y1] methy1/ ] benzidine [Q2] on the infection with Giardia lamblia. Treatment of Giardiasis 2 weeks post-infection with compound T1, C1, orally administered gave a very highly significant reduction in the number of cysts/gm stool were the percentage reduction rate reported for compound T1 given in a dose of [100 and 60 mg/kg] was 94.3% and 83.8%] respectively. However groups treated with compound C1 given in a dose of [100mg/kg and 60mg/kg] resulted 96.2% and 89.1%. When Q1 and Q2, were administered orally a significant reduction in the number of cysts/gm stool was noticed. Percent reduction reported for compound Q1 given in a dose [80 and 60mg/kg] was 89.6% and 72.6% respectively. Representing a highly significant reduction in number of cysts/ gm stool. Where in the compound Q2 percent reduction of cysts/ gm stool was 80.35%, 68.9% stool. The effect of compounds T1, C1, Q1, Q2 on the vegetative [Trophozoite] forms in the small intestine of sacrificed hamsters was studied. Compound T1 orally administered in a dose [100 mg/kg and 60 mg/kg] gave a highly significant reduction in the number of trophozoites [96.7% and 91.7%]. However compound C1 resulted to a significant reduction in the number of trophozoites [75.2% and 60.2%]. It was found that in treatment with compound Q1 and Q2 [80mg/kg and 60mg/ kg] the former was much more efficient than the later i.e there is a highly significant difference [94.5%, 96.7%] in case of Q1, Q2 at a dose of 80mg/kg, where as groups treated with 60mg/ kg of Q1 and Q2 showed much less reduction in the number of trophozoites. The curative effect of these compounds almost ranks with metronidazole. Histopathological examination, revealed a profound effect on the structure and function of the intestinal mucosa, villous shortening and a trophy, hypercellularity of the lamina propria due to increase in the number of mononuclear, polymorphonuclear and eosinophilic cells with diffuse loss of brush border microvillus surface area

Immunolocalization of schistosoma mansoni and schistosoma haematobium antigens reacting with their Egyptian snail vectors

The reaction of the haemolymph and the tissue of infected intermediate hosts, Biomphalaria alexandrina and Bulinus truncatus to Schistosoma mansoni and S. haematobium antigens were investigated using the indirect immunoperoxidase technique. A new technique, Agarose cell block was used in collection of haemolymph which helped in collecting plenty of well formed cells in comparison to the ordinary one using the cytospin. Collected haemolymph and prepared tissues of uninfected and infected B. alexandria and B. truncatus were fixed and then reacted with anti-S. mansoni and anti-S. haematobium IgG polyclonal antibodies. The haemolymph and tissue of infected B. alexandrina and B. truncatus gave a positive peroxidase reaction represented by a brown colour. In haemolymph, the positive peroxidase reaction was detected mainly in the cytoplasm of the amoebocytes. In the tissue, it was detected in epithelial cells lining the tubules, male cells in the lumen of the tubules and in female oogonia cells along the periphery of the tubules. The similarity in the strength and distribution of positive reaction in B. alexandrina and B. truncates was observed as compared to control. Thus, the immunoperoxidase technique proved to be an effective indicator for the schistosome-antigen in the snails

Circulating soluble Fas and Fas antigen tissue expression in human bladder cancer

This study was performed to determine the role of sFas and Fas expression in human bladder cancer. The percussion of schistosomiasis association and correlations between various parameters and tumor progression were evaluated as well. Fifty patients [24 with chronic cystitis and 26 with bladder cancer [20 with transitional cell carcinoma and 6 with squamous cell carcinoma]] were included in this study and fifteen individuals served as normal controls. SFas level in sera was estimated using enzyme linked immunosorbent assay. Fas expression in bladder tissue was immunohistochemically determined. SFas level and% of Fas expression in chronic cystitis and bladder cancer patients were significantly higher than normal controls. Moreover, a significant increase in sFas level and% of Fas expression was detected in chronic cystitis associated with schistosomiasis compared to chronic nonspecific cystitis. SFas level was significantly increased with tumor progression in the invasive group compared to the noninvasive group; whereas,% of Fas expression was comparable in both groups. Moreover, the number of Fas positive cases was significantly high in invasive than noninvasive

Expression of intercellular adhesion molecule ICAM-1 and cytokeratin as Potential markcrs of disease activity in the Egyptian pattern of chronic hepatitis C

Chronic hepatitis C [ch.HCV] and schistosomal hepatic fibrosis or both as a mixed hepatic lesion [MHL] are among the most common causes of endemic chronic hepatic disease in Egypt. Adhesion molecules especially ICAM-1 play an important role in inflammatory and immunological responses of chronic liver disease. Cytokeratin 18 [CK-18], although normally expressed in hepatic tissue, yet it is altered during chronic inflammatory hepatic lesions. This work was planned to study ICAM-1 as expressed in hepatic tissue in the different grades of the disease activity, in relation to its circulating levels in patients sera, and to evaluate the level of CK-18 expression in relation to the different grades of chronic inflammation and stages of fibrosis in the examined liver biopsies. The material for this study comprised 33 patients [17 ch.HCV and 16 MHL]. Seven cases, that proved to have nearly normal serological data and insignificant histopathological hepatic features, were considered as controls. All patients were assessed for HCV serological markers as well as serum levels of soluble ICAM-1 [sICAM-1] by the Enzyme Linked Immunosorbent. Assay [ELISA]. Liver needle biopsy specimens were processed and assessed for the histopathological grade of the disease activity and stage of fibrosis of the hepatic lesion. Tissue expression of ICAM-1 and CK-18 was detected using immunohistochemical techniques. Our results revealed significantly higher levels of serum sICAM-1 in both ch.HCV and MHL groups compared to controls [p<0.01]. Meanwhile, higher levels of sICAM-1 were recorded in the MHL cases relative to ch.HCV cases. ICAM-1 expression was not detected in any of the control cases, while it was positively expressed in all ch. HCV and MHL cases, with a higher score recorded in the later group [P>0.001 compared to the control group]. ICAM-1 expression was detected mainly within the sinusoidal cells [endothelial and Kupffer cells], hepatocytes, mononuclear inflammatory cells and vascular endothehail cells in portal areas. On classifying patients according to their grades of active inflammation and stages of fibrosis, higher scores of ICAM-1 expression within the hepatic tissue were recorded in cases with more active inflammatory grades and higher fibrotic stages. On the other hand, serum of ICAM-1 levels though were significantly elevated in patients with higher grades of inflammatory activity yet, they were decreased in patients with higher fibrotic stages. CK18 expression was mainly detected within hepatocytes of the periportal areas [in a combined membranous and intracytoplasmic pattern], as well as within the bile ducts epithelium in the portal areas. Over expression of CKI 8 was detected in both the ch.HCV and MHL groups [p<0.001 relative to controls], but the expression scores were higher in the MHL group. From these results we may conclude that MHL is a more aggressive and active chronic inflammatory hepatic disease than ch.HCV alone. Also, serum levels of sICAM-1 as well as hepatic expression of both ICAM-1 and CK-18 are related to the degree of disease activity, which may point out to the possibility of using serum sICAM-1 levels as well as the expression scores of hepalic ICAM-1 and CK-18 as efficient tools for monitoring the disease activity in ch. HCV and MHL patients. While ICAM-1 expression in tissue could be used as indicator for the stage of fibrosis in those patients also recommend that both ICAM-1 in serum andhepatic tissue could be used for monitoring the effect of therapy on the studied pattern of chronic hepatitis C