| Objective:Hepatic fibrosis is a pathologic process that leads to deposition of an excess of fibrous connective tissue, which is due to the synthesis of fibrous connective tissue exceeding the degradation of fibrous connective tissue. The balance between synthesis and degradation of fiber has been broken. Multiple etiological factors cause injury and inflammation of liver. Hepatic fibrosis is an important pathologic character in most kinds of chronic liver disease, which induce hepatic cirrhosis. In some circumstance, hepatic fibrosis was reversible .But, if the etiological factors keep on, hepatic cirrhosis will aggravate gradually. At last, hepatic lobules and vascular structures will be reconstructed, the normal hepatic tissues will be destructed, central vein region and header region will be separated, and will be formed, that is to say the cirrhosis has been appeared which is not reversible. Many cells, cytokines and changes of cellularmatrix in hepatic tissues are associated with hepatic fibrosis. Among them, HSCs are"activated"to a myofibroblast-like phenotype plays a central role in the development and resolution of liver fibrosis. Various factors associated with fibrosis are all targeted at HSCs which could be proliferated and transformed to myofibroblast resulting in hepatic fibrosis. In our country, HBV and HCV are the common entities of hepatic fibrosis, 20%-30% chronic hepatitis C patients eventually will show up hepatic fibrosis or hepatic cirrhosis.In the western country, chronic alcoholism is the main cause among which 60%-70% will result in cirrhosis. The research on the mechanism, prevention and cure of the hepatic fibrosis has become one of the hottest problems in the biologic field and medical science research both in home and aborad. At present,the treatment for hepatic fibrosis is till in the exploration stage, and specific drugs can not be got. Chinese medicine is showing its specific advantages in treating hepatic fibrosis. Its research has been furthered into molecular level of cells. Minority has been furthered into the level of intracellular information proventling. Its mechanism of action are multiple links,multipe way and multiple targets. Curcumin, a plant-derived polyphenol withdrew from curcuma, whose clinical properties of anti-tumor, anti-oxidant, and antiinflamm- ation, anti-human mim-unodeficiency virus, and live rprotection need further confirmation. This article summarizes the recent- advances in the experiment research and clinical application of curcumin, especially its role in the treatment of digestive diseases.This study used intraperitoneal injection with CCl4 establish a mouse model of hepatic fibrosis, application curcumin intervention therapy as positive provent. With method of immuno-histochemistry and RT-PCR to investigate the curative effect of curcum on hepatic fibrosis and express of caspase-3 in hepatic tissue of mice. Intervention HSC with different concentration of curcumin to investigate the depressant effect, precipitate HSC apoptosis, prevent the progress of hepatic fibrosis. To reveal the anti-hepatic fibrosis effect of Chinese medicine further, and provide new ideas for prevention and treatment of hepatic fibrosis effectively.Methods: healthy female C57BL6/J mice were randomly divided into 4 groups: normal group,prevention group,treatment group and model group,each with 10. All mice except normal group were given 10% CCL4 olive oil (5ul/g) by intraperitoneal injection,two times a week for 8 weeks. Normal group was given 0.9% Sodium Chloride by intraperitoneal injection, two times a week for 8 weeks. Curcumin (50 ug/g)was given by intragastric administration to prevention group and treatment group, three times a week for 8 weeks at beginning of 1st and 7th week. All groups mice were killed at 8th week. Mice serumal ALT were measured by automatic biochemical analyzer. HA were easured by intellectual radioimmunityγ-measurement appearance. Pathological changes in hepatic tissue were observed by HE and Masson staining. The expression ofα-SMA,caspase-3 of in hepatic tissue were examined by immunohistochemistry. The expression of caspase-3 mRNA were analyzed by RT-PCR. Affter treatment with curcumin for 24h, The cycle and apoptosis of HSC-T6 cells were studied by flow-cytometry (FCM). Results: 1. The common behavior of mice: normal mice were active, their hair was bright and weight increased gradually. Body weight of mice in model group decreased remarkably compared with normal group. Hair, appetite and body weight of mice in prevention group were much better than model group. More time of treatment with curcumin, symptoms of treatment mice were more better. The difference of body weight of mice in model,prevention and treatment group decreased compareing with normal group was significant(P<0.05);The difference of body weight of mice in prevention and treatment group increased compareing to model group was significant(P<0.05). There was no difference between prevention and treatment group(P>0.05).2. The examination of biochemical markers of serum: the levels of serum ALT of mice: model group>treatment group> prevention group> normal group(168.62±13.75 U/L,104.86±11.68 U/L,82.98±9.57U/L,47.91±8.08 U/L,P<0.05), the difference was significant;The levels of serum HA of mice: model group>treatment group> prevention group> normal group(1609.95±125.91ng/ml,1290.26±127.59ng/ml,953.39±98.48ng/ml,759.10±77.31ng/ml,P<0.05),the difference was significant.3. Pathological changes in the liver: Liver histology of normal group was changed rarely. was found in model group. There were lobules of liver,severe cellular necrosis and obvious fibrosis can be seen in model group. Large areas of mixed inflammatory infiltration with single nuclear cell can be seen throughout the hepatic lobular. The extent of inflammation, cellular necrosis and fibrosis of liver in prevention group and treatment group were dramatically decreased, and prevention group was better than treatment group.4. The expression ofα-SMA in hepatic tissue:there were only few expression in vessel wall in normal group. In model group,overexpression ofα-SMA in vessel wall, collects the district and fiber structure can be seen.Compared with model group, the expression ofα-SMA in the liver of prevention group and treatment group were decreased. The expression ofα-SMA in hepatic tissue in every group were: model group> treatment group> prevention group> normal group(8.21±1.48,6.25±1.69,4.78±1.32,1.51±0.95,P <0.05) the difference was significant.5. The expression of caspase-3 protein in hepatic tissue: there were only few expression in Disse space, central veins wall in normal group; The expression of caspase-3 protein was increases obviously in model group, in cytolymph,central veins,fibrosis district and Disse space can be seen. Compared with model group, the expression of caspase-3 in the liver of prevention group and treatment group were decreased. The expression ofα-SMA in hepatic tissue in every group were: model group> treatment group> prevention group>normal group(9.88±1.32,7.91±1.28,2.62±0.89,0.62±0.17, P<0.05), the difference was significant.6. The expression of caspase-3 mRNA in hepatic tissue: experimental mice hepatic tissue caspase-3 mRNA expression levels were: model group>treatment group>provention group> normal group. (0.441±0.013,0.374±0.010,0.310±0.010,0.203±0.006,P<0.05),the difference was significant.6.After treatment with curcumin, HSC-T6 cells shown some typical morphologic features, including cell shrinkage, cytoplasm concentration, nclius forming outwardly acute angle promineney, chromatin concentrating on karyotheca or forming meniscus, nuclear condensation, nuclear fragmentation and formation of apoptotic bodies. The result of FCM showed that curcumin can induce apoptosis and cell cycle of HSC-T6 cells. Affter treated by curcumin for 24h, the apoptosis rate of HSC-T6 increased markely with a dose-dependent manner. Meanwhile, after treatment with curcumin, the cell cycles of HSC-T6 cells were changed, the cells in G2/M phase were increased and cells in S phase were decreased.Conclusions:1.The hepatic fibrosis model could be established successfully by intraperitoneal injection with CCl4 to mouse for 8 weeks. And this model is consonant with the pathological features of human.2. Curcumin could improve hepatic function and hepar necrosis degree,have an contribution of treat hepatic fibrosis.3.Curcumin could inhibit hepatic tissue caspase-3 mRNA and protein expression changes, reduce hepatic fibrosis and liver tissue inflammation, prevent the progress of hepatic fibrosis and provides new insight to treatment of hepatic fibrosis. 4.Curcumin could inhibit growth of HSC-T6, inhibitory effect can be related to induce apoptosis of cells and arrest of cell cycle,curcumin could arrest HSC-T6 cell cycle at G2/M, the effect have an evident dose dependability. |
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