This study aimed to explore the effect of kushenin combined with adefovir dipivoxil on the load of hepatitis B virus DNA (HBV-DNA) in serum, in immune functions and in liver functions of patients with chronic hepatitis B. A sample of 80 patients with chronic hepatitis B was selected who were admitted to Weifang People's Hospital for treatment between January, 2013 and December, 2015. They were divided into the observation group (n=40) and the control group (n=40). The patients in both groups received adefovir dipivoxil, while those in the observation group additionally received the kushenin. Variations in HBV-DNA load and transforming growth factor-β1 (TGF-β1) in the two groups were detected before intervention, at 1 month, 3 months and 6 months after intervention. In addition, after intervention, we also observed the changes in CD4+, CD8+ and CD4+/CD8+, as well as the levels of immune globulin. Furthermore, in these two groups, we detected the changes in endotoxin in serum before and after intervention, the liver function after intervention, and the variations of hyaluronic acid (HA) and type III procollagen (PCIII) before and after intervention which were used to serve as the indicators for hepatic fibrosis. Results showed that at one month, 3 months and 6 months after intervention, HBV-DNA load and the level of TGF-β1 in the observation group were lower than those in the control group (P<0.05). In the observation group, the HBV-DNA load at 6 months after intervention was the lowest, sequentially followed by the levels at 3 months, at 1 month and before intervention (P<0.05). After intervention, the levels of CD4+ and CD8+ and CD4+/CD8+ in the observation group were higher than those in the control group (P<0.05). Moreover, the levels of immunoglobulin M (IgM), immunoglobulin G (IgG) and immunoglobulin A (IgA) were elevated in the observation group compared to the levels in the control group (P<0.05). Additionally, the level of endotoxin in serum, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBil). Besides, after intervention, the levels of HA and PCIII in the observation group were found to be lower than those in the control group before and after intervention (P<0.05). This study concludes that, for patients with chronic hepatitis B, kushenin combined with adefovir dipivoxil can remarkably decrease the HBV-DNA load, improve their immunity, ameliorate the liver function and delay the onset of liver cirrhosis.
Hepatitis B virus (HBV) is one of the most common DNA viruses at present which belongs to the Hepadnaviridae family of viruses. It is also considered as one of the most serious issues of social public health in China (
Currently, antivirus therapy has been the most common method for treatment of patients with hepatitis B. Comprehensive therapies including immunotherapy are also applied (
A total of 80 patients with chronic hepatitis B was selected who were admitted to Weifang People's Hospital for the treatment between January, 2013 and December, 2015. All patients underwent biochemistry examinations, and based on the results and on the medical history, we excluded the patients who were complicated with severe cardiorespiratory disorders, or had received therapies with the administration of immune preparations within one year before this study, or were allergic to the drugs used in this study, or were complicated with mental diseases. Furthermore, written signed informed consent was obtained from the patients, and this study was approved by the Ethics Committee of Weifang People's Hospital. Patients were divided into the observation group (n=40) and the control group (n=40) at random. The observation group consisted of 29 males and 11 females aged from 39 to 70 years (average age of 49.6±2.3 years). In addition, the disease course of hepatitis B was between 5 and 45 years with an average of 23.3±1.1 years. Furthermore, 13 patients were positive to hepatitis B surface antigen (HBsAg), hepatitis B virus e-antigen (HBeAg) and hepatitis B core antibody (HBcAb), and 27 patients were positive to HBsAG, hepatitis B virus e-antibody (HBeAb) and HBcAb. There were 21 patients with alcohol-drinking history. The control group consisted of 30 males and 10 females aged from 40 to 70 years (average age of 49.5±2.4 years). The disease course of hepatitis B was between 5 and 45 years with an average of 23.5±1.0 years. Moreover, 12 patients were positive to HBsAg, HBeAg and HBcAb, and 28 patients were positive to HBsAG, HBeAb and HBcAb. In this group, 20 patients had alcohol-drinking history. Comparison of gender, age, disease course and type of hepatitis B, and the alcohol-drinking history showed no statistically significant differences (P>0.05) between the two groups.
During the treatment, the progression in the disease course of patients was observed closely, and proper treatment method and support were provided for dealing with the specific symptoms. Regular examinations were carried out to detect the changes in the blood routine parameters, coagulation function, hepatic function and renal functions. Once patients were found with the levels of ALT and/or AST over 200 U/l, live-protective drugs were given immediately. The patients were withdrawn from administration of interferon and immunosuppressive agent as soon as they were enrolled into the study. In the control group, an oral administration of adefovir dipivoxil (q.d., 10 mg/day; NMPN: H20060666; Chia Tai Tianqing Pharmaceutical Group Co., Ltd., Jiangsu, China) was performed for fasting patients in the morning, where one course of treatment referred to the consecutive administration for 48 weeks. In the observation group, patients were given kushenin (t.i.d., 0.2 g/time; NMPN: 403405H28; Chia Tai Tianqing Pharmaceutical Group Co., Ltd.) in addition to the adefovir dipivoxil. Here one cycle of treatment consisted of two consecutive courses of treatment that was constituted by 24 consecutive weeks of treatment.
Variations in HBV-DNA load and transforming growth factor-β1 (TGF-β1) in the two groups were detected before intervention, at 1 month, 3 months and 6 months after intervention. In addition, after intervention, we also observed the changes in CD4+, CD8+ and CD4+/CD8+, as well as the levels of immune globulin. Furthermore, in both groups, we detected the changes in endotoxin in serum before and after intervention, the liver function after intervention, and the variations of hyaluronic acid (HA) and type III procollagen (PCIII) which served as the indicators for hepatic fibrosis before and after intervention.
The assay of HBV-DNA load was performed through fluorescence quantification polymerase chain reaction (PCR), in which the normal range was not >500 copy/ml, TGF-β1 level was detected via enzyme-linked immunosorbent assay (ELISA), the levels of endotoxin in serum was measured via limulus ameobatic chromogenic assay (normal reference value was set at 0.1 EU/ml). Relative indexes of liver functions including the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBil) were all measured using the Olympus AU2700 Automatic Biochemical Analyzer, in which the normal reference range in adults of TBil was between 3.4 and 20 µmol/l while the normal reference values of ALT and AST in adults were below 40 U/l. For cellular immune functions, the cell count of T lymphocyte subgroups, CD4+ and CD8+, was performed using the BD FRCS Calibur Flow Cytometer (BD Biosciences, Franklin Lakes, NJ, USA) automatically during which the supplementary reagents produced by BD were used, and 5 ml fasting elbow venous blood samples were collected in the morning and treated using ethylenediaminetetraacetic acid (EDTA) for later preservation. All the samples were delivered for examination within 12 h after removal from the body. For humoral immune functions, ELISA was performed for assay of immunoglobulin M (IgM), immunoglobulin G (IgG) and immunoglobulin A (IgA) in the body, in which the normal range of IgG in adults was set between 9.6 and 23.6 mg/l, the normal range of IgM was between 3.05 and 3.85 mg/l, and the normal range of IgA was between 3.14 and 4.66 mg/l. The levels of HA and PCIII were measured through solid-phase radioimmunoassay, in which the normal reference value of HA in adults was set as 47.6±22.5 ng/ml and the normal range of PCIII in adults was below 120 µg/l.
SPSS 13.0 provided by IBM (Chicago, IL, USA) was used to perform statistical analysis. The measurement data was presented as mean ± standard deviation. The comparison of means between the two groups was carried out using t-tests. In addition, the Chi-square test was performed for intergroup comparison of rates. Furthermore, intragroup comparisons at different time-points before and after intervention were made using the variance analysis of repeated measurements. For all statistical test, P<0.05 was considered as the statistical significance level.
Results showed (
The results showed (
After intervention, the levels of CD4+ and CD8+ in the observation group were higher compared to those of the control group (P<0.05), while on the other hand, CD4+/CD8+ was higher than that in the control group (P<0.05) (
The results (
Before intervention, no statistically significant difference was found in comparison of endotoxin level in serum between the two groups (P>0.05). However, after intervention, the level of endotoxin in serum in the observation group was significantly lower than that in the control group (P<0.05) (
The results show (
Before intervention, the level of HA in the observation group was 215.6±25.3 ng/ml, and the level of PCIII was 119.5±14.3 µg/l. After intervention, the HA level was 83.2±7.1 ng/ml, and the PCIII level was 60.3±5.1 µg/l. In addition, before intervention, the level of HA in the control group was 216.3±25.6 ng/ml, and the level of PCIII was 120.1±14.5 µg/l. After intervention, the HA level was 138.6±11.9 ng/ml, and the PCIII level was 86.6±6.8 µg/l. Furthermore, before intervention, there was no statistically significant difference in HA and PCIII between the two groups (P>0.05). However, after the intervention, the levels of HA and PCIII in the observation group were lower than those in the control group before and after intervention (P<0.05) (
Hepatitis B is an important factor that often causes hepatic cirrhosis. The onset of hepatic cirrhosis is usually accompanied by the decrease in urine volume (
As a new type of nucleotide reverse transcriptase inhibitor, namely the adefovir dipivoxil, was used in this study which can effectively antagonize a variety of viruses. It has been recommended by World Health Organization as the first-line drug for treatment of infections of hepatitis B and acquired immune deficiency syndrome. The efficacy on these diseases has been observed in clinical practice (
After the patients in the observation group received the combination treatment of kushenin and adefovir dipivoxil, the HBV-DNA load and TGF-β1 levels were significantly decreased, and the immune function was effectively improved, which may be correlated with the features of oxymatrine, the effective constituent of kushenin, such as the anti-inflammation effect and the capability to eliminate the free radicals (
This study concluded that for patients with chronic hepatitis B, kushenin combined with adefovir dipivoxil can significantly decrease the HBV-DNA load, ameliorate the immune function and liver function, and delay the onset of hepatic cirrhosis.
Comparison of hepatitis B virus DNA (HBV-DNA) load at different time-points between two groups before and after intervention.
Comparison of transforming growth factor-β1 (TGF-β1) levels at different time-points between two groups before and after intervention.
Comparison of the indicators of hepatic fibrosis [hyaluronic acid (HA) and type III procollagen (PCIII)] between the two groups before and after intervention.
Comparison of HBV-DNA load at different time-points before and after intervention between the two groups (copy/ml).
Group | Before intervention | In 1 month after intervention | In 3 months after intervention | In 6 months after intervention | F-value | P-value |
---|---|---|---|---|---|---|
Observation group | 16311.2±103.6 | 11326.2±89.9 | 327.2±32.8 | 213.5±21.5 | 191.092 | <0.01 |
Control group | 16315.3±104.3 | 12312.5±99.1 | 829.1±41.2 | 505.3±89.3 | 114.192 | <0.01 |
t-value | 0.163 | 46.621 | 24.248 | 6.321 | – | – |
P-value | 0.871 | <0.01 | <0.01 | <0.01 |
HBV-DNA, hepatitis B virus DNA.
Comparison of TGF-β1 level at different time-points before and after intervention between the two groups (pg/ml).
Group | Before intervention | In 1 month after intervention | In 3 months after intervention | In 6 months after intervention | F-value | P-value |
---|---|---|---|---|---|---|
Observation group | 128.6±21.0 | 100.2±18.1 | 81.2±12.4 | 63.3±8.9 | 11.292 | <0.01 |
Control group | 130.1±20.9 | 115.4±19.2 | 92.1±14.2 | 99.6±13.8 | 9.192 | <0.01 |
t-value | 0.320 | 3.643 | 3.657 | 13.981 | – | – |
P-value | 0.750 | <0.01 | <0.01 | <0.01 | – | – |
TGF-β1, transforming growth factor-β1.
Comparison of the variations in CD4+, CD8+ and CD4+/CD8+ levels after intervention between the two groups.
Group | CD4+ (mm3) | CD8+ (mm3) | CD4+/CD8+ |
---|---|---|---|
Observation group | 35.1±2.0 | 30.6±1.3 | 1.9±0.3 |
Control group | 26.0±0.9 | 24.4±1.0 | 1.6±0.2 |
t-value | 26.242 | 23.908 | 5.262 |
P-value | <0.01 | <0.01 | <0.01 |
Comparison of the variations in immunoglobulin after intervention between the two groups (mg/l).
Group | IgM | IgG | IgA |
---|---|---|---|
Observation group | 1.6±0.03 | 9.5±0.12 | 3.9±0.20 |
Control group | 1.3±0.02 | 8.8±0.10 | 1.8±0.15 |
t-value | 52.623 | 28.342 | 53.126 |
P-value | <0.01 | <0.01 | <0.01 |
IgM, immunoglobulin M; IgG, immunoglobulin G; IgA, immunoglobulin A.
Comparison of level of endotoxin in serum between the two groups before and after intervention (EU/ml).
Group | Before treatment | After treatment |
---|---|---|
Observation group | 1.31±0.3 | 0.06±0.01 |
Control group | 1.30±0.3 | 0.99±0.06 |
t-value | 0.149 | 96.967 |
P-value | 0.882 | <0.01 |
Comparison of the indexes related to liver functions after intervention between the two groups (mean ± SD).
Group | ALT (U/l) | AST (U/l) | TBil (µmol/l) |
---|---|---|---|
Observation group | 37.6±2.5 | 39.5±2.4 | 19.5±2.8 |
Control group | 60.6±5.9 | 59.8±6.3 | 29.8±4.6 |
t-value | 22.701 | 19.044 | 12.097 |
P-value | <0.01 | <0.01 | <0.01 |
ALT, alanine aminotransferase; AST, aspartate aminotransferase; TBil, total bilirubin.