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

Study of kinetics of circulating schistosome antigens in tissues and sera of infected mice using anti-schistosoma mansoni soluble egg antigen monoclonal antibody

In the present study, the kinetics of uptake and deposition of Schistosoma mansoni antigens in liver, spleen, kidney and intestine of C57BL/6 mice infected with 100 Schistosoma mansoni cercariae as well as their levels in sera have been investigated during the course of infection [12 weeks]. The presence of antigen was demonstrated by indirect immunofluorescence using IgM anti-soluble egg antigen monoclonal antibody [anti-SEA MAb]. Immunofluorescence reactivity was evident in both renal and spleen tissues 3 weeks post-infection [p.i.], in Kupffer cells of liver 4 weeks p.i. and in intestinal mucosa [5 weeks p.i.]. Maximal immunofluorescence staining was reached during the patent phase [5-9 weeks p.i.]. During the chronic stage of infection [9-12 weeks pi.], diminution of immunofluorescent intensity was evident in liver tissue, while it remained constant in other studied organs. Circulating schistosome antigen [CSA] level in mice sera was determined using a sandwich ELISA with a MAb for both antigen capture and detecting antibody. CSA was demonstrated in mice sera [one week p.i.] reaching its peak at 6 weeks p.i. and remained at a detectable level until the end of the study [12th week p.i.]

dynamics of serum and hepatic schistosomal antigens in two different strains of mice

This work was designed to study the dynamics of circulating schistosome antigens [CSA] both in serum and hepatic tissues to evaluate their role in the pathological changes in murine schistosomiasis mansoni in two strains of mice with different genetic make up [C57BL/6 and BALB/c] during the course of schistosomal infection. The detection of serum CSA was performed using sandwich ELISA. Immunolocalization of schistosomal antigens in hepatic tissue was studied using indirect immunofluorescent staining. In both techniques, an IgM anti-Schistosoma mansoni soluble egg antigen [SEA] monoclonal antibody [MAb] used. Immunolocalization as well as detection of CSA were studied weekly starting from the first week up to twelve weeks post-infection [p.i.]. The serum circulating antigen was detected as early as the first week p.i.; whereas, its detection in hepatic tissue was observed at a later stage [fourth week p.i.]. Serum circulating antigen levels reached their peaks at the 5th and 6th weeks p.i. in BALB/c and C57BL/6 mice, respectively, and then declined. The deposited schistosomal antigen in hepatic tissues reached their peaks at 7th- 8th week p.i. in both strains, but declined more rapidly in C57BL/6 mice than in BALB/c mice. The data denoted that the dynamics of circulating antigen both in the serum and the hepatic tissues varies according to the genetic background of the mice