| ObjectiveHepatitis B virus (HBV) infection is a worldwide epidemic and has become a serious public health problem that threaten human health. Studies suggest that hepatic inflammation is mainly caused by HBV infection-induced adaptive immunity, not by HBV itself. Immune pathogenesis is the main pathogenesis of chronic hepatitis B. Current therapies include anti-virus, immune regulation, liver function improvement and anti hepatic fibrosis. Antiviral agents for hepatitis B include antiviral nucleoside (nucleotide) analogs and interferon. But antiviral nucleoside (nucleotide) analogs can easily lead to virus mutation. Interferon, which has narrow adaptation and big side effects, is not only expensive, but also has poor long-term efficacy. And these antiviral agents has no effect on covalently closed circular DNA (cccDNA), which is the template for viral RNAs.so it is difficult to completely eleminate virus from a person’s body, leading to high rate of relapse after drug withdrawal.My tutor held the view that dampness-heat epidemic toxin attack the human body from qi to blood aspect because of lack of healthy qi, leading to liver depression and spleen deficiency which often manifest as abdominal fullness, torpid intake and lack of strength. Besides, liver depression and blood stasis can result in hypochondriac pain,bile obstruction can cause jaundice. Treatment of Chronic hepatitis B should be reinforce the healthy qi and eliminate the pathogenic factors. After years of clinical practice, he created a decoction called zhenqiyigan decoction. The whole decoction is composed of Astragalus, Ligustrum lucidum, lucid ganoderma, Hypericum japonicum, Caulis polygoni multiflori, Radix isatidis, etc. The decoction has an effect of reinforcing the healthy qi and enriching yin, clearing heat and drain dampness, detoxifying and tonifying qi. It has good clinical efficacy for chronic hepatitis B. In order to summarize and promote the decoction, clinical and experimental studies on the treatment of acute immune-mediated liver injury was carried out. In clinical studies, we evaluated the clinical effect of zhenqiyigan decoction through randomized controlled study, camparing with the western medicine group:entecavir tablets. In experimental studies, acute immunologic liver injury model was prepared by tail intravenous injection of concanavalin A.We explored the mechanism of the protective function of the decoction from apoptosis and necrosis and regeneration aspects of liver cells, respectively.MethodsClinical study:We observed 71 patients with chronic hepatitis B. Which were divided into treatment group of 40 cases and control group of 31 cases. Treatment group were treated with zhenqiyigan decoction which were given one prescription daily and taken 300ml each time,2 times a day, and control group were treated with entecavir tablets,0.5mg each time, once a day. Two groups were treated for 6 months. We analyzed symptom scores, liver function and virological indicators before and after treatment.Experimental study:120 KM female mice were randomly divided into six groups:high-, middle-, and low-dosage zhenqiyigan decoction groups, the bifendate group, normal control group and model group. Other groups were treated with concanavalin A (15mg/kg)by tail vein injection to make acute immunologic liver injury model except for normal control group. After modeling successfully, high-, middle-, and low-dosage zhenqiyigan decoction groups and the bifendate group were given corresponding drugs by gavage, and the model group and the normal control group with an equal volume of saline for 7 days. Three mice were taken daily from each group, blood and liver samples were taken and ELISA were used to detect serum levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), malondialdehyde(MDA), superoxide dismutase(SOD), interferon-γ (IFN-γ), tumor necrosis factor-a(TNF-a), interleukin-4(IL-4), interleukin-6 (IL-6) and interleukin-10 (IL-10). Expression levels of Fas, FasL, Bax, BCL-2 and PCNA in liver tissue were detected by immunohistochemistry method. Experimental results are indicated by mean±standard deviation, using repeated measures analysis of variance to compare the efficacy of each group, with P<0.05 as statistically significant difference.ResultsClinical study:Baseline data of the patients in two groups showed no significant difference in gender, age, medical history, symptom severity and symptom scores(P>0.05). Two groups could improve the patient’s clinical symptoms. In improving patients with stuffiness and fullness in the stomach, abdominal fullness, inability to sleep, profuse dreaming, viscous and inhibited stool and reddish yellow urine, the treatment group was better than the control group (P<0.05). Both the treatment group and the control group could promote serum HBeAg seroconversion, decrease the serum levels of HBV-DNA, improve liver function, and two groups showed no significant difference (P>0.05). In comparison of clinical efficacy, The total effective rate of treatment group was 55% and 51.6% in the control group, but there was no statistically significant difference between two groups(P<0.05). In improving the patient’s symptoms and signs, the total effective rate in the treatment group was 95% and 77.4% in the control group. the treatment group was better than the control group (P<0.05).Experimental Study:(1) Effect of zhenqiyigan decoction on serum levels of ALT and AST in miceCompared with normal control group, after treated with Con A, In model group, high-, middle-, and low-dosage zhenqiyigan decoction groups and the bifendate group, serum levels of ALT and AST was increased in different degrees(P<0.01). Compared with model group, high-, middle-, and low-dosage zhenqiyigan decoction groups and the bifendate group could reduce serum levels of ALT and AST, In reducing serum levels of AST, and high-dosage zhenqiyigan decoction group was superior to the bifendate group(P<0.05).(2)Effect of zhenqiyigan decoctionon on serum levels of MDA and SOD in mice.Compared with normal control group, after treated with Con A, In model group, high-, middle-, and low-dosage zhenqiyigan decoction groups and the bifendate group, serum levels of MDA increased(P<0.01)and SOD reduced in different degrees(P<0.01).Compared with model group, high-, middle-, and low-dosage zhenqiyigan decoction groups and the bifendate group could reduce serum levels of MDA and SOD, and high-dosage zhenqiyigan decoction group was superior to the bifendate group (P<0.05) in the decrease in serum level of S0D(P<0.05).(3) Effect of Zhenqiyigan decoctionon on serum levels of TNF-a, IFN-y, IL-4, IL-6 and IL-10 in miceCompared with normal control group, after treated with Con A, In model group, high-, middle-, and low-dosage zhenqiyigan decoction groups and the bifendate group, serum levels of TNF-α, IFN-γ and IL-6 increased and IL-4 and IL-10 reduced in different degrees(P<0.01). Compared with model group, high-, middle-, and low-dosage zhenqiyigan decoction groups and the bifendate group could reduce serum level of TNF-α, IFN-γ and IL-6 and increase IL-4 and IL-10, In addition to IFN-y, the high-dosage group was better than bifendate group (P<0.05).(4) Effect of Zhenqiyigan decoction on expression levels of Fas, FasL, Bax, BCL-2 and PCNA in liver tissueCompared with normal control group, after treated with Con A, in model group, high-, middle-, and low-dosage zhenqiyigan decoction groups and the bifendate group, expression levels of Fas, FasL, Bax, PCNA increased and BCL-2 reduced in different degrees(P<0.01). Compared with model group, high-, middle-, and low-dosage zhenqiyigan decoction groups and the bifendate group could increase expression levels of Fas, FasL, Bax, PCNA and reduce BCL-2. High-dosage group was better than bifendate group in expression levels of PCNA (P<0.05).(5)pathological manifestations of liver tissueIn normal control group, central vein could be seen in the center of the hepatic lobule, and hepatic plate composed of hepatocyte in the middle of the lobule. Between the hepatic plate was liver sinusoid. In model group.large areas of necrosis could be seen in the hepatic lobule, around it was the infiltration of inflammatory cells, the structure of the hepatic lobule was damaged. In the high-dosage zhenqiyigan decoction group and bifendate group, hepatic cells swelled obviously which was called ballooned changes in hepatic lobule. In the middle-dosage zhenqiyigan decoction, spotty necrosis could be seen in hepatic lobule, around it was the infiltration of inflammatory cells. In the low-dosage zhenqiyigan decoction, spotty necrosis could be seen around central vein in hepatic lobule, around it was the infiltration of inflammatory cells.ConclusionClinical Study:Zhenqiyigan decoction can improve the clinical symptoms and signs and liver function of patients with chronic hepatitis B, with a higher seroconversion rate, it can be effective in the treatment of chronic hepatitis B.Experimental Study:(1)After treated with Con A, serum levels of ALT and AST in mice was increased significantly after 8 hours. It indicates that acute immune-mediated liver injury model was successfully made.(2)After the application of zhenqiyigan decoction, compared with model group, serum levels of ALT, AST in mice was significantly decreased, indicating that zhenqiyigan decoction has protective effect on immunological liver injury model and can decrease serum levels of transmigrate.(3)After the application of zhenqiyigan decoction, compared with model group, serum levels of MDA and SOD in mice was decreased and increased accordingly, indicating that zhenqiyigan decoction has the effect of antioxidant and alleviating liver injury.(4)After the application of zhenqiyigan decoction, compared with model group, not only serum levels of TNF-a, IFN-y and IL-6 was decreased and IL-4 and IL-10 increased accordingly, but also pathological changes alleviated, indicating that zhenqiyigan decoction has the effect of alleviating liver injury by down-regulating proinflammatory cytokines and up-regulating anti-inflammatory cytokines to regulate the balance of th1/th2.(5)After the application of zhenqiyigan decoction, compared with model group, not only expression levels of Fas, FasL, Bax and PCNA increased, but also Bcl-2 decreased, indicating that zhenqiyigan decoction plays a protective role of promoting the damaged liver cell apoptosis and stimulating regeneration of liver cells. |
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