| Purpose To study the pathological changes of liver fibrosis, and to explore the activation of hepatic stellate cells(HSCs), the dynamic changes in the expression of nuclear factor kappa B (NF- B), transforming growth factor beta-1 ( TGF- 1) and type I receptor of transforming growth factor beta (TR 3 I ) in the development of rat hepatic fibrosis and the effects of Compound Astragalus Extract (CAE) on them. Methods SD rats were divided into six groups: normal control, model control, groups at three doses ( 60 , 120 , 240 mg-kg-1 ) CAE and Liver-Aid tablets(921mg-kg-1). CCl4 (12.5%, 4ml-kg-1) was injected sub-cutaneously in rat twice a week for 8 and 13 weeks. The drug at three different doses (0.5% CMC-Na in control groups) was given intragastricly once a day for 8 and 13 weeks. The rats were killed at the end of 8th and 13th week respectively, the left liver tissue was then used to make tissue microarrays (TMA). The TMA sections were used for hematoxylin and eosin( HE ) stain , Van Giesons ( VG ) collagen stain , reticular fiber stain , Immunohistochemistry (IHC) and In situ hybridization (ISH ). Imrnunohistochemical S-P method was used to detect the expression of a -smooth muscle actin ( a -SMA) and proteins of NF-KB p65, TGF- 1 and TR I . mRNA expression of TGF- P i and TRP I was detected by ISH. a -SMA positive cells were counted in three random 250nm 250 m areas under light microscope ( 200). The expression of NF- B protein, TGF- 1 protein and its mRNA, T R I protein and its mRNA were quantifiedby MetaMorph imaging analysis system. Results 1. During 8 and 13 weeks, the degrees of liver injury and fibrosis grades of hepatofibrotic rats liver induced by CCl4 of model group were higher than those in normal group ( 8 weeks, P<0.01 ; 13 weeks, P<0.001 ), which were amelionated remarkably by CAE ( 60~240mg-kg-1 ) treatment. 2. The number of activated hepatic stellate cells (HSCs) in fibrotic model liver tissue was more than that in normal group during 8 and 13 weeks (P<0.001 ). The protein expression of NF- K B p65 , the protein and mRNA expression of TGF- P and T P R I in fibrotic model liver tissue were stronger than those in normal group during 8 and 13 weeks (P<0.001 ). The number of activated HSCs, protein expression of NF- K B p65, protein and mRNA expression of TGF- P i and T P R I were positively correlated to each other during 8 weeks, while the number of activated HSCs was only correlated with the expression of NF- K B p65 protein (PO.01 ) and TGF- P i mRNA (PO.001 ) during 13 weeks. The expression of NF- K. B p65 protein, protein and mRNA expression of TGF -P i and T 3 R I were positively correlated to each other during 13 weeks(PO.001 ). 3. CAE (60- 240mg-kg"1 ) inhibited the activation and proliferation of HSC of fibrotic liver in rats during 8 and 1 3 weeks ( CAE at low and high doses group during 8 weeks and CAE at middle dose group during 13 weeks, P<0.01 ; CAE at low and high doses group during 13 weeks and CAE at middle dose group during 8 weeks, P<0.001 ); CAE (60~240mg-kg-1) inhibited the expression of NF- K B p65 protein of fibrotic liver tissue in rats during 8 and 13 weeks (CAE at low dose group during 8 weeks, P<0.05; the other CAE doses group during 8 and 13 weeks,PO.001 ); CAE ( 60~240mg-kg"' ) dose-dependently inhibited not only the protein expression of TGF- P i and T P R I during 8 weeks(r TGF-P1i Protein= -0.683 , r TPR i protein= -0.796, PO.001 )but also the protein expression of TGF- 1 during 13 weeks (r TGF-P i protein= -0.629, PO.001) of fibrotic liver tissue in rats, but it only had a tendency to inhibit the expression of T P R I protein(P>0.05) of fibrotic liver tissue in rats during 13 weeks. CAE (60-240mg-kg"1) inhibited the mRNA expression of TGF- P i and T P R I of fibrotic liver tissue in ratsduring 8 and 13 weeks ( CAE at low and high doses group during 13 weeks, P<0.01 ; the other CAE doses group during 8 and 13 weeks, .P<0.001). Conclusion A model for liver fibrosis may be established by subcutaneous injection of 1... |
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