| Objectives: ⑴This study takes WD liver fibrosis patients with no difference in age and course as the research object,and chooses Danshen pills as the comparative treatment method,to observe the function of Gandoufumutang(GDFMT)with patients’ score about their UWDRS liver function,TCM syndrome integral value and the value of TGF-β1,TNF-α、IL-6 and to evaluate the clinical efficacy of GDFMT about treating WD liver fibrosis patients.⑵ This research is based on the TGF-β1 signal transactional pathway,taking WD TX model mice as the research object and GDFMT as the treatment drug.It uses the semi-quantitative rt-pcr to detect TGF-β1,TβRI,TβRII,Smad2,Smad3,Smad4 and Smad7 after the interference of the high,medium and low concentration levels of GDFMT.It discusses the curative effects of GDFMT fight WD of liver fibrosis and its possible molecular mechanisms.Methods: ⑴ Clinical research part: Sixty patients with WD liver fibrosis meeting the criteria were randomly divided into two groups,thirty patients for the treatment group and others for the control group.Treatment group used GDFMT while control group used salvia miltiorrhiza particles.The foundation treatment with DMPS of the 2 groups were same.Eight days was counted as a course,during which six days was the copper vein row treatment and the remaining 2 days to be calcium.There were four sessions in total.The differences between the two groups before and after the treatment were compared through measuring TCM syndrome integral value,the UWDRS liver function value and the values of TGF-β1,TNF-α、IL-6.⑵ Laboratory experiment part: 192 male TX(saline)mice were randomly assigned to the model group,the penicillamine group,the danshen group,the GDFMT low-dose group,the GDFMT dose group,the GDFMT high-dose group,and 32 DL mice as the normal control group,seven groups in total.All of them were given the corresponding drugs,once every day and 56 days continuously.At the end of the second,fourth,sixth and eighth weeks,the experimented mice were killed in batches,eight at a time in each group.In the end,the semi-quantitative rt-pcr was used to test the relative expression quantity of TGF-β1,TβRI,TβRII,Smad2,Smad3,Smad4 and Smad7 in the mice liver tissueResults:(1)The clinical research part: Clinical efficacy:After four courses of treatment,the clinical total effect of treatment group DMPS joint GDFMT is superior to control group DMPS and salvia miltiorrhiza group;the syndrome integral value of two groups of patients before treatment had no difference(P> 0.05);the UWDRS liver function score values of two groups of patients before treatment had no difference(P > 0.05);the TCM syndrome integral values of two groups after treatment were lower than before treatment(P< 0.01,P< 0.05);Compared with treatment groups before,two groups’ UWDRS liver function score values were lower after(P< 0.01,P< 0.05);The TCM syndrome integral value of treatment group decreased significantly than the control group after the treatment(P< 0.01);The treatment group’s UWDRS liver function score reduced more significantly than the control group(P< 0.01).The validity evaluation of clinical efficacy: Comparing TGF-β1、TNF-α、IL-6 of treatment group and control group before treatment,the three index had no significant difference(P> 0.05);Comparing TGF-β1、TNF-α、IL-6 of treatment group after treatment and before treatment,the three index decreased significantly(P< 0.01).Comparing TGF-β1、TNF-α、IL-6 of the control group after and before the treatment,the three index decreased remarkably(P< 0.01,P< 0.05);Comparing the treatment group and the control group after the treatment,the treatment group improved more obviously than the control group(P< 0.01).It can be concluded that GDFMT is better anti-fibrosis effects.Laboratory experimental part The comparison results of the inspection index by treatment of GDFMT after 2,4,6,8 weeks are as follows:(1)After 2,4,6,8 weeks,TGF-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA,had a significant rise in value,Smad7 m RNA value decreasing significantly(P< 0.01)in the model group compared with the normal group;(2)The comparison between each treatment group and their corresponding model groups are as follows: TGF(1)-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA value decreased,Smad7 m RNA value increased in high dose group,middle dose group,danshen group,penicillamine group after 2,4,6,8 weeks and the low dose group after 4,6,8 weeks compared to the corresponding model groups,(P< 0.05,P< 0.01).(2)There is no statistical difference between the low dose group after week 2 and the model group(P> 0.05);(3)GDFMT high dose group,low dose group,and its corresponding GDFMT salvia miltiorrhiza group,penicillamine group are as follows:(1)TGF-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA value decreased significantly,the relative expression of Smad7 m RNA increased(P< 0.05,P< 0.01)in GDFMT middle and low dose group,danshen group,penicillamine group after week 2,4,6,8 and penicillamine after week 4,6,8 compared with corresponding GDFMT high-dose group;compared(2)with penicillamine group 2 weeks,there is no statistical difference in the GDFMT high dose group(P> 0.05).(4)GDFMT dose group and salvia miltiorrhiza group,penicillamine group,low dose group are as follows:(1)compared with salvia miltiorrhiza group,there is no statistical difference in the dose group(P > 0.05);(2)compared with penicillamine group,medium dose group of 2,4 week of TGF-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA expression are relatively high compared with its quantity,the relative expression of Smad7 m RNA its lower(P< 0.05,P< 0.01);penicillamine group compared with dose group of 6,8 weeks,TGF-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA expression decreased relatively,the relative expression of Smad7 m RNA increased(P< 0.05,P< 0.01);(3)low dose compared with corresponding dose group of 6,8 weeks,TGF-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA comparative expression quantity is relatively low,the relative expression of Smad7 m RNA is high(P< 0.01);low dose group 2 weeks compared with the corresponding dose no difference(P> 0.05).(5)danshen group compared with penicillamine group,low dose group are as follows: danshen group 2,(1)4 weeks the corresponding penicillamine group TGF-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA expression relatively high compared with its quantity,the relative expression of Smad7 m RNA its lower(P< 0.05,P< 0.01);danshen group 6,8 weeks the corresponding penicillamine group TGF-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA expression decreased relatively,the relative expression of Smad7 m RNA increased(P< 0.05,P< 0.01);(2)danshen group 6,8 weeks compared with corresponding low dose group,TGF-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA expression quantity compared with its relatively low,the relative expression of Smad7 m RNA its high(P< 0.01);danshen group 2 weeks no difference compared with corresponding low dose group(P> 0.05).(6)penicillamine group and low dose group are as follows: penicillamine 2,4 weeks compared with corresponding the low dose group,TGF-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA expression quantity compared with its relatively low,the relative expression of Smad7 m RNA its high(P< 0.05,P< 0.01);penicillamine group 6 week the corresponding low dose group had no difference(P> 0.05);penicillamine 8 week is corresponding to the lower dose group,TGF-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA expression relatively high,relative expression of Smad7 m RNA lower(P< 0.01).(7)(1)GDFMT high dose group compared with the corresponding 2 weeks,4 weeks,TGF-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA relative lower,Smad7 m RNA relative higher(P< 0.01);(2)GDFMT middle and low dose group compared with the corresponding 2 weeks,4 weeks has no obvious difference(P> 0.05).(8)compared with corresponding 4 weeks,6 weeks TGF-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA relative lower,Smad7 m RNA relative higher(P< 0.01).(9)compared with corresponding to 6 weeks,8 weeks,TGF-β1、TβRI、TβRII、Smad2、Smad3 and Smad4 m RNA relative expression reduced,the relative expression Smad7 m RNA is higher(P< 0.01).Conclusion:(1)GDFMT can effectively improve TCM syndrome integral of WD patients in liver fibrosis,UWDRS liver function score and the clinical symptoms.⑵GDFMT can effectively lower TGF-β1,TNF-α,IL-6 of WD patients with liver fibrosis,and therefore plays the role of anti fibrosis.⑶GDFMT can effectively lower TGF-β1,TβRⅠ,TβRⅡ,Smad2,Smad3 and Smad4,and can effectively improve Smad7.GDFMT can inhibit the TGF-β1 / Smad signal transduction path and activate the HSC through the control of the protein expression,after which the anti-fibrosis effect can be approached.Moreover,it has ageing dependencies on the dose-effect of TGF-β1/Smad signal transduction. |
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