| Objectives To evaluate the effects of Baihe Wuyao Decoction(BWD)on treatment of chronic liver injury and liver fibrosis induced by carbon tetrachloride(CCl4),and to elucidate the possible underlying mechanisms.Methods Sixty eight-weeks Kunming male mice were randomly divided into six groups:normal group(CNTR),model group(Model),positive group(Positive),Baihe Wuyao decoction low-dose group(LD),middle-dose group(MD),and high-dose group(HD).Except CNTR and Model group,the mice in other groups were given by gavage once a day for successive 6 weeks.Among them,the mice in the positive group was given Diammonium Glycyrrhizinate at a dose of 0.585 g/10 g bodyweight,and LD,MD and HD of BWD were respectively administered at the doses of 0.05 g,0.15 g,0.45 g crude drug/10 g bodyweight.At the same time,except CNTR group,the mice in all other groups were intraperitoneally injected of 20% CCl4 dissolved in olive oil at a dose of 0.1 m L/10 g bodyweight twice a week.After the administration of 6 weeks,all the mice were fasted for12 h,and the serums were collected through the abdominal aorta and liver samples collected.The content of Alanine aminotransferase(ALT)and Aspartate aminotransferase(AST)in the serum were evaluated using automatic biochemical detector;The hepatic pathological changes and the collagen fibers distributions in the liver were observed through HE staining and Sirius scarlet staining;The antioxidant capacity of liver tissues were detected by the contents of superoxide dismutase(SOD)and malonaldehyde(MDA)in liver homogenate;The protein level expressions of collagen fibers Collagen1,Collagen3,alpha smooth muscle actin(?-SMA),transcription growth factor(TGF-?1),inducible nitric oxide synthase(i NOS),p-Smad2/3,and Smad2/3 in the liver were by analyzed using Western-blot;The m RNA level expressions of liver matrix metalloproteinases(such as MMP2,MMP9,MMP12),tissue inhibitor metalloproteinase(TIMP1),manganese superoxide dismutase(Mn-SOD),inducible i NOS,interleukin(IL-1?,IL-11),and tumor necrosis factor(TNF-?)were detected through quantitative Real-time PCR(RT-PCR).Results 1 Compared with the CNTR group,the mice in Model group exhibited that: 1)The liver weight index(the ratio of liver weight to body weight)was increased;There were many white granules on the surface of liver,and the liver morphologies were dramatically defected and severe liver fibrosis had been occurred;The serum ALT and AST contents were significantly increased,which indicated that liver functions were severely damaged;2)Western-blot results showed that the protein expressions of ECMs(such as Collagen1 and Collagen3)and the HSCs activated marker ?-SMA in the liver increased,which demonstrated that HSCs were activated;3)The expressions of TGF-?1,p-Smad2/3,Smad2/3 in the liver were upregulated which indicated that TGF-?1?Smad signal pathway was activated;4)The gene expressions of MMPs and TIMP1 were increased,which suggested that increased TIMP1 inhibited the degradation of MMPs on Collagens;5)The expressions of MDA and i NOS in the liver increased,while the expression of Mn SOD and SOD decreased,which indicates that the ability of scavenging free radicals in the liver was weakened.6)The expression of IL-1?,IL-11 and TNF-? in the liver increased,indicating that the liver of the mice had occurred inflammatory response.2 Compared with the Model group,the mice in BWD groups exhibited that: 1)Almost no white particles were observed on the surface of the livers;The liver morphology and fibrosis were significantly improved;In addition,the content of ALT and AST in serum decreased and the liver function was obviously ameliorated;2)The expressions of Collagen1,Collagen3 and ?-SMA decreased in the liver,and at the same time,HSCs activities were suppressed;3)The liver TGF-?1?Smad2/3 signal pathway was inhibited;4)The expressions of MMPs in the liver were up-regulated,while the expression of TIMP1 was down-regulated,indicating that the degradation of MMPs for ECMs was enhanced;5)The expressions of MDA and i NOS in the liver were elevated,while the expressions of Mn SOD and SOD were decreased,suggesting that the antioxidant capacity of the liver was enhanced;6)The expressions of IL-1? ? IL-11 and TNF-? were reduced,which manifested that liver anti-inflammatory activity were increased.Conclusions BWD has a good protective effect on chronic liver injury and liver fibrosis induced by CCl4,which underlying mechanisms are related to its blocking of TGF-?1?Smad2/3 signaling pathway,anti-inflammatory effects and antioxidant effects.Figure 11;Table 6;References 144... |
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