Effect Of A Curcumin Derivative On Rat Liver Fibrosis Induced By CCL₄

Objective To evaluate the efficacy of Curc-OEG in preventingCCl₄-induced by hepatic fibrosis.Methods （1） Establishment of rat hepatic fibrosis model:85SD malerats were randomly divided into6groups: group A（n=10）, normal control;group B（n=15）, CCl₄control; group C（n=15）, D（n=15） and E（n=15） wereCCl₄and curcumin derivative （25,50,100mg.kg-1.d-1） treated rats; groupF（n=15） was treated with CCl₄and colchicin as positive control. All ratswere fed with normal diet. Except that those in group A were givensubcutaneous injection of CCl₄and olive oil mixture （2:3）.The dosage of2mL/kg were given twice a week for10weeks except that the first dose was5mL/kg.（2） Examine that effect and the relative mechanism of Curc-OEG onrat liver fibrosis induced by CCl₄: After4weeks of modeling, those ingroup A and B were treated with normal saline by intravenous injection oncea day, those in group C, D and E were treated with Curc-OEG for25,50,100mg.kg-1.d-1by intravenous injection, those in group F were treated withcolchicin for0.1mg.kg-1.d-1by gavages. During the experiment, all rats were weighed before giving the mixture of CCl₄and olive oil to adjust thedose of CCl₄. At the end of10thweek after Curc-OEG administration, ratswere anesthetized with ether and blood was obtained from the heart forserum biochemistry analysis. Samples from the right liver lobes were thenfixed in10%formaldehyde solution for histological analysis containing HEand Sirius red staining. Samples from the left liver lobes were immediatelyfrozen for determination of matrix metalloproteinase-13, tissue inhibitor ofmetalloproteinase-1，collagenâ…  and hydroxyproline contents.Results Compared with model group, the serum levels of ALT andAST was obviously decreased in those treated groups of25,50,100mg.kg-1.d-1and colchicin. Compared with normal group, all groupsexcept high-dose group were increased. These differences were statisticallysignificant （P<0.05）. The contents of Hyp in all groups given CCl₄wereobviously increased in comparison with normal group. Compared withmodel group, the contents of Hyp in treated groups of50,100mg.kg-1.d-1and colchicin were reduced except for low-dose group. Compared withlow-dose group, the contents of Hyp in medium-dose group and high-dosegroup were a remarkable reduction and colchicin group was no statisticalsignificance. But the content of Hyp in medium-dose group and high-dosegroup was obviously lower than colchicin group, and high-dose group waslower than medium-dose group. HE and Sirius red staining showed that theliver tissue structure was normal, no inflammatory cells, no fibrosis proliferation. The marked bridging was observed in portal to portal andportal to central, and the part of these were observed nodules occasionallyin model group. But collagenous fiber accumulation were reduced in thosegroups of Curc-OEG treatment with25,50,100mg.kg-1.d-1and colchicintreatment, and the higher the dosage, the better effect. Results weremeasured in terms of%area occupied by specific staining and these werescored as described previously.Compared with model group, the gene transcription of MMP-13wasup-regulated and the gene transcription of TIMP-1was down-regulated inmedium-dose group, high-dose group and colchicin group except that thegene transcription of MMP-13and TIMP-1were no difference in theexpression of low-dose group. In addition, the expression ofMMP-13mRNA in medium-dose group, high-dose group was higher thanlow-dose group but the expression of TIMP-1mRNA in medium-dosegroup, high-dose group was lower than low-dose group. The expression ofMMP-13mRNA and TIMP-1mRNA were no statistically significantdifference between low-dose group and colchicin group. Compared withmedium-dose group, the expression of MMP-13mRNA was up-regulatedand the expression of TIMP-mRNA was down-regulated in high-dosegroup. The expression of Collagenâ…  of model group was obviously higherthan normal group by RT-PCR and Western blot. The expressions ofCollagenâ… in those treatment groups of medium-dose group, high-dose group and colchicin group were lower than model group and low-dosegroup, and low-dose group was not lower than model group. But comparedwith colchicin group, the expression of Collagenâ… in group low-dose wasno statistical significance. And medium-dose group and high-dose groupwere lower than colchicin group in expression of Collagenâ… . And thehigh-dose group was lower than medium-dose group in expression ofCollagenâ… .Conclusion Curc-OEG could ameliorate the progress of liver fibrosisinduced by CCl₄.