| Objective Hepatitis B is a common chronic infectious disease in China,which seriously threatens the lives and health of Chinese residents.There is currently no ideal therapeutic drug,and exploring new drugs remains a top priority.The Melastoma dodecandrum Lour is used in the treatment of hepatitis in the private sector,but there is still no experimental basis for the herbal anti-viral and liver-protecting effects.This study intends to evaluate the anti-duck hepatitis B virus effect and liver protection efficacy of the water extract and alcohol extract of the herb through animal models,and provide a scientific basis for the development of new drugs for the treatment of hepatitis B.Methods 1.A total of 201 1-day-old Guangxi ducks were purchased,24 of which were subjected to short-term repeated dose toxicity test,8 in each group,divided into Melastoma dodecandrum Lour water extract group（3.64 g/kg·d）,and alcohol extract group（3.64 g/kg·d）,normal control group（normal saline）.After 28 days of repeated oral administration,the water extraction and alcohol extraction were evaluated for the short-term use of the toxic properties and target organs of the ducks.2.After the remaining 177 1 day old Guangxi ducks were fed for 24 hours,155 of them were intraperitoneally injected with 0.15 ml of duck hepatitis B virus serum.The jugular vein blood was collected on the 7th day,and the serum duck hepatitis B virus DNA（DHBV DNA）was detected by PCR.22 were not treated,and serum DHBV DNA-negative ducks were screened.3.Randomly selected 80 serum DHBV DNA positive ducks were divided into model group（normal saline）,lamivudine group（200 mg/kg·d）,high（3.64g/kg·d）,medium（1.82 g/kg·d）and low（0.91 g/kg·d）dose group of water extract,and high（3.64 g/kg·d）,medium（1.82 g/kg·d）and low（0.91 g/kg·d）dose group of alcohol extract,each group of 10.10 of the serum DHBV DNA negative ducks were randomly selected for the experimental blank group（normal saline）.A total of 9 experimental groups.The drug was administered once a day in the morning from the 14th day after the injection of the virus serum,and for 28 consecutive days.Before administration（T0）,on the 7th day of administration（T7）,on the14th day of administration(T14),on the 21st day of administration(T21),on the28th day of administration(T28),and 5th day after drug withdrawal（P5）,collecting jugular vein blood separately.The ducks were sacrificed on the 6th day after stopping the drug,and the liver tissue samples were taken.4.Real-time quantitative PCR was used to detect serum DHBV DNA load by SYBR Green I fluorescent dye method,and its specificity,sensitivity and reproducibility were evaluated.And analyze the standard curve.The level of serum DHBsAg/DHBeAg was detected by enzyme-linked immunosorbent assay（ELISA）.Automatic biochemical analyzer for detecting serum ALT/AST activity.Finally,the liver tissue of ducks was pathologically examined.5.Data compilation and statistical analysis were performed using Excel2007 and SPSS20.0 software.The difference was statistically significant at P<0.05.Results 1.The effect of drugs on the levels of DHBsAg and DHBeAg:Lamivudine group,high,medium and low dose group of water extract/alcohol extract of Melastoma dodecandrum Lour compared with model group,there were no statistically significant difference（P>0.05）.2.The effect of drugs on DHBV DNA load:The serum DHBV DNA load of T7,T14,T21,T288 and P5 in lamivudine group was significantly different from that in the model group（P<0.01）,and rebound phenomenon occurred after stopping drug（P5）.There were no significant differences in serum DHBV DNA load between T7,T288 and P5 in the high,medium and low dose groups of water extract/alcohol extract（P>0.05）.3.The effect of drugs on alanine aminotransferase（ALT）activity:The serum ALT activity of T7 in the high dose group of water extract was lower than that in the model group（P<0.05）.The serum ALT activities of T14,T21,T288 and P5 in the high,middle and low dose groups of water extract/alcohol extract were lower than those in the model group（P<0.05 or P<0.01）.There was no significant difference in T7 serum ALT activity between the high dose group of water extract and the blank group（P>0.05）.There was no significant difference in T288 serum ALT activity between the high dose group of water extract,high dose group of alcohol extract and the blank group（P>0.05）.The serum ALT activities of T211 and T288 in the high,medium and low dose groups of water extract/alcohol extract were lower than those before administration（T0）（P<0.05or P<0.01）.The serum ALT activity of the high,medium and low dose groups of water extract and the high and middle dose groups of alcohol extract were still lower than T0 after stopping the administration（P<0.05 or P<0.01）,but slightly higher than T28（P<0.05）.4.The effect of drugs on aspartate aminotransferase（AST）activity:The AST activity of the lamivudine group,the high dose group of water extract and the high dose group of alcohol extract in the T288 serum was lower than that in the model group（P<0.05）.There was no significant difference between the high,medium and low group of water extract/alcohol extract and the model group in other treatment time（P>0.05）.5.The main types of pathological changes in duck liver tissue were inflammatory cell infiltration in the portal area,loose cytoplasm and spot-like necrosis,with statistically significant differences in the degree of liver tissue lesion among the experimental groups（x2=30.476,P<0.01）.The pathological changes of duck liver tissue in the lamivudine group,the high dose group of water extract and the high dose group of alcohol extract were significantly different from those in the model group（P<0.05）.The pathological changes of liver tissue in the high dose group of water extract were lighter than those in the low dose group（P<0.05）.The pathological changes of liver tissue in the high dose group of alcohol extract were lighter than those in the medium dose group and the low dose group（P<0.05）.Conclusion 1.There is no obvious anti-duck hepatitis B virus effect in the water extract and alcohol extract of Melastoma dodecandrum Lour.2.The water extract and alcohol extract of Melastoma dodecandrum Lour have protective effects on liver injury caused by DHBV infection in ducks. |
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