The Influence Of Bushen Huoxue Huazhuo Recipe On The Noninvasive Diagnosis Model Of Wilson’s Disease Liver Fibrosis And The Liver Regeneration Of TX Mice Liver Fibrosis

1 ObjectiveBy observing the effect of Bushen Huoxue Huazhuo Recipe on APRI,FIB-4 and GPR index of Wilson disease hepatic fibrosis,we can provide the evaluation basis for the clinical treatment effect.Meanwhile,By observing the effect of Bushen Huoxue Huazhuo Recipe on TGF-β₁,BMP-7 of liver issue of TX mice,we can investigate the mechanism of liver regeneration in Wilson disease TX mice with liver fibrosis.2 Methods2.1 Clinical researchFrom January 2019 to December 2019,50 WD patients with liver fibrosis were randomly divided into control group and treatment group,25 cases each group.The control group was treated with sodium dimercaptosulfonate injection,10%calcium gluconate,zinc gluconate tablets and glutathione.On the basis of conventional western medicine treatment,the treatment group was treated with Bushen Huoxue Huazhuo Recipe,which was composed of Radix Rehmanniae Praeparata 20g,Schisandra 6g,Morinda 15g,Rhubarb 8g,Lysimachia 15g,Alisma orientalis 9g,Coptis 9g,Salvia miltiorrhiza15g,Zedoary 9g,Turmeric 10g and so on.Four items of liver fiber,liver function,platelet count and APRI,FIB-4,GPR were observed before and after treatment in the two groups.2.2 Experimental researchTX mice were randomly divided into model control group,Bushen huoxue huazhuo fang group,Penicillium group and glutathione group after 1 week of adaptive feeding TX mice and 10 mice DL 4 months old.In the group of Bushen Huoxue Huazhuo Recipe,on the basis of gandouling tablet,add the traditional Chinese medicine for tonifying the kidney（Radix Rehmanniae Praeparata 20g,Schisandra 6g,Morinda15g and so on）,and dissolve it in the normal saline,and each mouse is given gavage according to 6.90g/kg/d;Each mouse in penicillamine group was given penicillamine tablets as 0.15g/kg/d according to the literature,and each mouse in glutathione group was given glutathione tablets as 0.18g/kg/d.The blank group and model group were given the same amount of normal saline once a day for 4 weeks.Finally,the content of hydroxyproline in the liver tissue of mice was detected by hydroxyproline kit,the m RNA expression of TGF-β₁and BMP-7 in the liver tissue was detected by RT-PCR,and the protein expression of TGF-β₁and BMP-7 in the liver tissue was detected by Western blot.3 Results3.1 Clinical research3.1.1 Clinical efficacy:After four courses of treatment,the difference of the clinical efficacy between the two groups was statistically significant（P<0.01）.The clinical efficacy of treatment group was better than control group.3.1.2 Four liver fibers were compared between the two groups before and after treatment.PⅢNP、CⅣThere was no significant difference in the levels of HA,LN,PⅢNP and CⅣbetween the two groups before treatment（P>0.05）;After 4 courses of drug treatment,the levels of HA,LN,PⅢNP and CⅣdecrease in the control group and the treatment group（P<0.05）;The level of HA and CⅣin the treatment group was better than that in the control group（P<0.05）;There was no significant difference in LN and PⅢNP between the treatment group and the control group（P>0.05）.3.1.3 ALT,AST,GGT and PLT before and after treatment were compared between the two groups.There was no significant difference in ALT,AST,GGT and PLT between the two groups before drug treatment（P>0.05）.After 4 courses of drug treatment,the levels of ALT and AST in the control group and the treatment group decreased（P<0.05）;The levels of PLT in the control group and the treatment group increased;There was significant difference in AST between the treatment group and the control group（P<0.01）.There was no significant difference in ALT,GGT and PLT between the treatment group and the control group（P>0.05）.3.1.4 APRI,FIB-4 and GPR before and after treatment were compared between the two groups.There was no significant difference in APRI,FIB-4 and GPR between the two groups（P>0.05）;After 4 courses of drug treatment,the levels of APRI and FIB-4 in the control group and the treatment group（P<0.05,P<0.01）;the levels of GPR in the treatment group also decreased（P<0.05）;The level of APRI and FIB-4 in the treatment group was superior to the control group（P<0.01）.3.2 Experimental research3.2.1 Determination of hydroxyproline in mouse liverThe Hyp content in the model control group was significantly higher than the blank control group,（P<0.01）;Compared with model control group,the content of Hyp in liver tissue of TX mice in Bushen Huoxue Huazhuo Recipe group,penicillamine group and glutathione group decreased significantly（P<0.01）.3.2.2 Detection of TGF-β₁and BMP-7 m RNA expression in mouse liver by RT-PCRCompared with the blank control group,the expression of TGF-β₁m RNA in the liver tissue of TX mice of the model control group was significantly higher（P<0.01）,the expression of BMP-7 m RNA in the model control group was significantly lower（P<0.05）.Compared with the model control group,the expression of TGF-β₁m RNA in the liver tissue of TX mice in Bushen Huoxue Huazhuo Recipe group,penicillamine group and glutathione group were significantly lower,while their BMP-7 m RNA were significantly higher（P<0.01,P<0.05）3.2.3 Detection of TGF-β₁and BMP-7 protein expression in mouse liver by Western-BlotCompared with the blank control group,the expression of TGF-β₁protein in the liver tissue of TX mice of the model control group was significantly higher（P<0.01）,the expression of BMP-7 protein in the model control group was significantly lower（P<0.05）.Compared with the model group,the expression of TGF-β₁protein in the liver tissue of TX mice in Bushen Huoxue Huazhuo Recipe group,penicillamine group and glutathione group were significantly lower,while their BMP-7 protein were significantly higher（P<0.01,P<0.05）.4 Conclusion（1）Bushen Huoxue Huazhuo Recipe can reduce the four items of liver fibrosis and the serum indexes of liver function in patients with Wilson disease.（2）Bushen Huoxue Huazhuo Recipe can reduce the index of Apri and FIB-4 liver fibrosis non-invasive diagnosis model in patients with Wilson disease.（3）Bushen Huoxue Huazhuo Recipe may be through promoting the expression of BMP-7,inhibiting the expression of TGF-β₁,reducing the generation of collagen fibers,and playing an anti fibrosis role by regulating the process of liver regeneration.