| Objective : The subject objective to observe the clinical efficacy and safety of "Ji Sheng Shen Qi" decoction combined with entecavir in the treatment of patients with hepatitis B cirrhosis due to deficiency of spleen and kidney yang,and itseffects on liver function and bonemarrow hematopoiesis.to observe the molecular mechanism of "Ji Sheng Shen Qi" decoction combined with bone marrow mesenchymal stem cells transplantation in hepatic cirrhosis rats,and to explore theeffect of Jishengshengqi decoction,a prescription of tonifying kidney and marrow,on promoting stem cell migration.The theoretical basisof traditional Chinese medicine for improving liver function is related to modern molecular biological mechanism.Methods:(1)Clinical study:64 patients with hepatitis B cirrhosis with deficiency of spleen and kidney yang were randomly divided into treatment group(n = 32)and control group(n = 32).The control group was treated with entecavir dispersible tablets,the treatment group was treated with entecavir dispersible tablets combined with "Ji Sheng Shen Qi" decoction for 3 months.After the treatment,the changes of clinical symptom score,liver function,blood routine,serum erythropoietin(EPO)and adverse reactions were observed before and aftertreatment.(2)Animal experiment:77 healthy SPF grade SD rats were randomly divided into 3 rats after being killed,bone marrow mesenchymal stem cellss were taken out for cell culture,10 rats were used as normal group,the other rats were treated with carbon tetrachloride combined with ethanol.The 8W cirrhotic rat model was induced by compound factors of high fat diet.The cirrhotic rats were randomly divided into model group,single BMSCs transplantation group,single Chinese medicine group,BMSCs transplantation combined with Chinese medicine high dose group,BMSCs transplantation combined with Chinese medicine medium dose group,BMSCs transplantation combined with Chinese medicine low dose group.The blank group was fed with distilled water,the model group was given no intervention,the BMSCs group was given BMSCs transplantation,the traditional Chinese medicine group was given the medium dose of "Ji Sheng Shen Qi" decoction suspension.BMSCs transplantation combined with high dose Chinese medicine group was given BMSCs transplantation and high dose gastric perfusion of "Ji Sheng Shen Qi" decoction suspension,BMSCs transplantation combined with middle dose Chinese medicine group was given BMSCs transplantation and "Ji Sheng Shen Qi" decoction suspension medium dose intragastric perfusion;BMSCs transplantation combined with low dose Chinese medicine group was given BMSCs transplantation and "" Ji Sheng Shen Qi " decoction" suspension low dose gastric perfusion.The Chinese medicine was given once a day,BMSCs was transplanted once a week,from the end of the 8th week to the end of 12 weeks.After the treatment,the histopathology of the liver was observed in 7 groups,and the liver function(ALT,AST,ALB,TBIL)was measured by automatic biochemical analyzer.The level of EPO,GCSF,IL-3,IL-6,HGF,SCF in peripheral blood of rats in each group was detected by ELISA method.The expression of SDF-l,CXCR4 protein in liver tissue of rats in each group was detected by;Western blot method.Results:(1)Clinical study:In the course of the experiment,3 cases were eliminated because the patients did not take drugs according to the treatment plan,the patients lost the visit and the patients automatically asked to withdraw because of personal factors.The final effective case was 61 cases.After the course of treatment,the clinical symptom score,liver function,blood routine and serum erythropoietin were improved after treatment in the treatment group(P<0.01),and the clinical symptom score and liver function in the control group after treatment were improved(P<0.01).The blood routine was better than that before treatment(P<0.01),but the serum erythropoietin in the control group was not significantly different from that before treatment(P>0.05).The clinical symptomscore,liver function,blood routine and serum erythropoietin in the treatment group were better than those in the control group(P<0.01 or P<0.05).There were no side effects and side effects in the two groups.(2)Animal experiment:The general situation of rats:after the intervention treatment,the spirit,nutrition and activity of the rats in each treatment group were improved in varying degrees compared with those before the intervention treatment.Among them,BMSCs transplantation combined with high dose Chinese medicine group and BMSCs transplantation combined with Chinese medicine medium dose group had the best effect on improving the spirit,nutrition and activity of rats.The changes of HE staining in liver tissue of rats in each group:the pathological changes of liver tissue in model group were severe fibrosis,pseudolobules could be seen,and the degree of degeneration and necrosis of liver cells was serious,which indicated that the model of hepatic cirrhosis was successful.After intervention,the liver fibrosis,degeneration and necrosis of livercells in each treatment group were improved to some extent.Among them,BMSCs transplantation combined with high dose Chinese medicine group and BMSCs transplantation combined with Chinese medicine medium dose group hadthe best effect on improving liver fibrosis,hepatocyte degeneration and necrosisin cirrhotic rats,BMSCs transplantation combined with traditional Chinese medicine with low dose and single Chinese medicine transplantation group had the second effect.All of them were superior to the traditional Chinese medicine group alone.Liver function: compared with the blank group,the levels of ALT,AST and TBIL in the model group were significantly higher than those in the control group,and the level of ALB was significantly decreased(P<0.01).Compared with the model group,the levels of ALT,AST and TBIL in each treatment group were decreased to some extent,and the level of ALB was increased in different degrees(P<0.01).There was no significant difference in the reduction of ALT,AST between the BMSCs transplantation group and the BMSCs medium dose group(P>0.05).The effect of increasing ALB,and decreasing TBIL was the best in BMSCs transplantation combined with high dose Chinese medicine group,followed by BMSCs transplantation combined with Chinese medicine medium dose group(P<0.01).Changes of EPO,GCSF,IL-3,IL-6,HGF,SCF in peripheral blood of rats in each group:compared with the blank group,the levels of EPO,G-CSF,HGF,SCF and IL-3 in the model group decreased significantly,and the level of IL-6 increased significantly(P<0.01).Compared with the model group,the levels of EPO,G-CSF,HGF,SCF and IL-3 in each treatment group were increased to varying degrees,and IL-6 levels were decreased in different degrees((P<0.01).Compared with each treatment group,the effect of BMSCs transplantation combined with high dose of traditional Chinese medicine on increasing EPO and reducing IL-6 was the best,followed by BMSCs transplantation combined with medium dose of Chinese medicine(P<0.01 or P<0.05).However,the effect of BMSCs transplantation combined with high dose of Chinese medicine was the best(P<0.01).The effect of),BMSCs transplantation combined with low dose of traditional Chinese medicine was better than that of BMSCs transplantation combined with medium dose of Chinese medicine.The difference was not statistically significant(P>0.05).The expression of SDF-l,CXCR4 protein in liver tissue of rats in each group: after intervention,the expression of SDF-l,CXCR4 protein in liver tissue of each treatment group was higher than that of model group(P<0.01).The expression of SDF-l,CXCR4 protein in liver tissue was the highest in BMSCs transplantation combined with Chinese medicine high dose group and BMSCs transplantation combined with Chinese medicine medium dose group,followed by BMSCs transplantation combined with Chinese medicine low dose group and single BMSCs transplantation group.All of them were higher than that of traditional Chinese medicine group(P<0.01).Conclusion: The clinical study showed that Enticavir combined with "Ji Sheng Shen Qi" decoction was effective,safe and reliable in the treatment of hepatitis B cirrhosis patients with spleen-kidney yang deficiency syndrome,and could obviously improve its clinical symptoms,liver function,blood routine and serum EPO level,promote bone marrow hematopoiesis.Animal experiments showed that BMSCs transplantation combined with "Ji Sheng Shen Qi" decoction could improve liver function,liver inflammation,liver fibrosis,promote bone marrow hematopoiesis and BMSCs migration.This may be related to the up-regulation of the SDF-l/CXCR4 signaling pathway and the promotion of the secretion of EPO,SCF,G-CSF,HGF and IL-3 downstream factors in the signal pathway,and the inhibition of IL-6 secretion in the downstream signal pathway.It provides a preliminary basis for the treatment of hepatic cirrhosis with Chinese medicine combined with BMSCs transplantation in the future. |
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