RESUMO
OBJECTIVE:To study the effect and potential mechanism of the total flavonoids from Marchantia convoluta on anti-hepatic fibrosis in the mice. METHODS :Seventy-two mice were randomly divided into blank group ,model group ,positive control group (colchicine 0.2 mg/kg)and M. convoluta total flavonoids high-dose ,medium-dose and low-dose groups (300,150, 75 mg/kg),with 12 mice in eac group. Except for blank group ,other groups were subcutaneously given 25% CCl4-peanut oil solution on the back to induce liver fibrosis model. At the same time ,blank group and model group were given water intragastrically,while other groups were given relevant medicine intragastrically 20 mL/kg,once a day ,for consecutive 10 weeks. After last administration ,the serum levels of ALT and AST were detected . Histopathological changes of liver tissue in mice was observed. The levels of COL- Ⅰ,COL-Ⅲ and TGF-β1 in liver tissue were detected . The protein expression levels of α-SMA and TGF-β1,Smad2,Smad4 and Smad 7 in liver tissue were detected . The expression levels of TGF-β1,Smad2,Smad4 and Smad 7 mRNA in liver tissue were detected . RESULTS :Compared with blank group ,the serum levels of ALT and AST in model group,the levels of COL- Ⅰ,COL-Ⅲ and TGF-β1 in liver tissue,protein expression levels of α-SMA,TGF-β1,Smad2 and Smad 4,mRNA expression levels of TGF-β1,Smad2 and Smad4 were increased significantly (P<0.05 or P<0.01).The mRNA and protein expression levels of Smad 7 in liver tis sue were decreased significantly (P<0.05). The degree of liver tissue injury and collagen fiber hyperplasia were serious. Compared with model group ,above indexes of mice were reversed significantly in positive control group and M. convoluta total flavonoids high-dose group (P<0.05 or P<0.01). Serum level of ALT ,the levels of COL- Ⅰ,mRNA and protein expression of TGF-β1,Smad2 and Smad 4 in liver tissue were decreased significantly in M. convoluta total flavonoids medium-dose group (P<0.05 or P<0.01). Protein expression of Smad 2 and Smad 4 in liver tissue were decreased significantly in M. convoluta total flavonoids low-dose group (P<0.05). The liver injury and fibrosis of mice were relieved in administration groups. CONCLUSIONS :M. convoluta total flavonoids possess the effect of anti-hepatic fibrosis ,the mechanism of which is related to the regulation of mRNA and protein expression of TGF-β1,Smad2,Smad4 and Smad 7 in the signaling pathway of TGF-β/Smad.
RESUMO
OBJECTIVE:To establish a host specific method for the qual ity evaluation of Taxillus chiueusis by evaluating the quality of T. chiueusis from different hosts sources. METHODS :HPLC was adopted with CAPCELL PAK C 18 MGⅡ column, mobile phase consisted of methanol- 0.2% phosphoric acid ,volume flow 1.0 mL/min(gradient elution ),detection wavelength of 254 nm,column temperature of 30 ℃. Totally of 16 batches of T. chiueusis from different hosts sources were collected as sample (4 batches from tea host ,maple host ,willow host and mulberry host respectively ). HPLC characteristic chromatogram was established with TCM Chromatogram Fingerprint Similarity Evaluation System (2004A edition ),and similarity evaluation was performed. The peak was identified by comparison with the reference substance. IBM SPSS 19.0 software was used for cluster analysis and principal component analysis. RESULTS :Totally 11 common peaks were demarcated ,peak No. 8,10,11 were identified as rutin ,quercetin and quercitin. The similarity of 16 batches of samples was more than 0.9. The chemical components of T. chiueusis from different hosts sources were basically the same (RSDs of relative time of commom peaks were 0.03%-0.40%), but the contents of the same component were quite different (RSDs of relative peak area of commom peaks were 27.00%-64.20%). By cluster analysis ,16 batches of T. chiueusis from different hosts sources were divided into 3 types. The results of principle component analysis showed that quercetin ,quercitin and rutin had significant effect on the quality of T. chiueusis ,which could be used as the quality evaluation index ,and the whole quality of maple host was the best (the highest comprehensive score ). CONCLUSIONS: HPLC fingerprint combined with chemometrics can be used to evaluate the quality of T. chiueusis from different hosts sources.