Effect Of Oral Entecavir Combine With Prednisone On Mice With HBV Infection

Objective：To detect the effect of entecavir and prednisone on HBVreplication of mice in vivo.To investigate whether the antiviral combine withprednisone can enhanced virus replication express in HBV replication mice.Methods：1.Detection of the effect of prednisone on HBV replication of miceHBV replication competent plasmid pAAV/HBV1.2was transferred intomale BALB/C mice by hydrodynamics-based in vivo transfection.24h aftertransfection, the mice were matched by body weight and serum level ofHepatitis B e antigen (HBeAg) and were divided into experiment group andcontrol group. The experiment group were treated orally with prednisonewhile the control group were treated with normal saline.The experimentgroup and control group were respectively killed at the3、7、10、16、22daysafter the administration.HBV-DNA in serum were analyzed by real-timePCR.(the lower limit value of HBV-DNA in serum is500copies/ml) Theserum HBsAg and HBeAg were detected by chemiluminescent detectionsystem.2. Detection of the effect of entecavir on HBV replication of miceHBV replication competent plasmid pAAV/HBV1.2was transferred intomale BALB/C mice by hydrodynamics-based in vivo transfection.24h aftertransfection, the mice were matched by body weight and serum level ofHepatitis B e antigen (HBeAg) and were divided into experiment group andcontrol group. The experiment group mice were treated orally with entecavir while the control group were treated with normal saline.The experimentgroup and control group were respectively killed at the1、3、6、11days afterthe administration.HBV-DNA in serum were analyzed by real-time PCR.Theserum HBsAg and HBeAg were detected by chemiluminescent detectionsystem.3. Detection of the effect of entecavir and prednisone on HBVreplication of miceHBV replication competent plasmid pAAV/HBV1.2was transferred intomale BALB/C mice by hydrodynamics-based in vivo transfection.24hoursafter transfection, the mice were matched by body weight and serum level ofHepatitis B e antigen (HBeAg) and were divided into6experiment groupsand3control groups. Every mouse of the4experiment groups were treatedwith orally1.5μg entecavir,and respectively treated with orally400μgprednisone at the same or1、3、and7days after entecavir administration. Theanother2experiment groups were treated with orally prednisone at the day of24hours after transfection and were treated with orally entecavir after3or7days of prednisone administration. The3control groups were treated withentecavir、prednisone、normal saline respectively.All of the mice werekilled at the16th day after prednisone be used.HBV-DNA in serum wereanalyzed by real-time PCR.The serum HBsAg and HBeAg were detected bychemiluminescent detection system.Result:1. Effect of prednisone on HBV replication of miceHBV-DNA in serum from mice of the prednisone experiment groups andnormal saline control group of3、7、10days were all positive,and the levelof serum HBV-DNA averages all reached to105copies/ml showing no significant difference;HBV-DNA in serum of prednisone experiment groupand normal saline control group of16days were1.83×105copies/ml、3.07×103copies/ml respectively.The level of experimental mice is higher thanthat of control mice.HBV-DNA in serum of22days prednisone experimentgroup and normal saline control group were showing significant differencetoo,their averages out to2.03×104copies/ml,1.99×103copies/ml.HBsAg in serum of the experiment and control groups of3、7、10daysmade no significant difference； The level of HBsAg in the serum ofexperiment group was higher than that of control group of16days,and theaverags out to53.35IU/ml、1.97IU/ml respectively; The level of HBsAg in theserum of the experiment group mice of22days was higer than that of controlgroup mice,too.HBeAg in serum of the experiment and control groups of3、7、10days showing no significant difference；But16days after treatment,theHBeAg level in serum of experiment mice was higher than that of controlmice;Similarly of22days after treatment, the HBeAg level in serum of theexperiment mice was higher than that of control groups.2.Effect of entecavir on HBV replication of miceHBV-DNA in serum of entecavir experiment group and normal salinecontrol group at the1th day were all positive,and the averages both reachedto104copies/ml,showing no significant difference.HBV-DNA in serum ofentecavir experiment group mice(103copies/ml) was lower than that of normalsaline control group(105copies/ml) of3days. HBV-DNA in serum ofentecavir experiment mice of6days were one negative and two positivereached to102copies/ml,and that of normal saline groups were all positiveaveraged to5.27×105copies/ml higher than the experiment group.HBV-DNAin serum of the entecavir experiment mice of11days were all negative,but that of normal saline control mice average reached to104copies/ml,the levelof control mice was higher than that of experiment mice.HBsAg and HBeAg in the serum of experiment and control groups of1、3、6、11days made no significant difference.HBeAg in the serum ofexperiment and control groups at the1th、3th、6th、11th days showde nosignificant difference.3.Effect of entecavir and prednisone on HBV replication of mice①HBV-DNA in the serum of normal saline control group averaged outto2.1×103copies/ml.HBV-DNA in the serum of the4experiment groupstreated with entecavir advanced and then prednisone were allnegative.HBV-DNA in the serum of experiment group which treated withprednisone advanced and then entecavir after3days averaged out to2.21×103copies/ml,making no significant difference with the normal salinecontrol group.HBV-DNA in the serum of experiment group which treatedwith prednisone advanced and then entecavir after7days averaged out to1.29×104copies/ml,while the level were higher than the normal saline controlgroup and the experiment group treated with prednisone advanced thenentecavir after3days.②HBsAg in the serum of the4experiment groups that treated withentecavir first and then prednisone were all lower than the normal salinecontrol group,showing no significant difference between groups. HBsAg inthe serum of the2experiment groups that treated with prednisone first andthen entecavir were all lower than the normal saline control group,and nosignificant difference was showed between groups. HBeAg in the serum ofthe4experiment groups that treated with entecavir firstiy and then prednisonewere all lower than the normal saline control group,the level lower the longer entecavir treated.HBeAg in the serum of the2experiment groups that treatedwith prednisone first then entecavir were all lower than the normal salinecontrol group.The level that treated with entecavir in7days latter were higherthan the3days latter.Conclusions：①The replication of hepatitis B virus in the HBV infection mice whichwere treated with prednisone orally could be restrained by treated withentecavir orally in advance or at the same time.②The activation of hepatitis B virus by prednisone in the HBV infectionmice couldn’t be restrained by entecavir orally in3days later.The more timelatter,the more significantly HBV were activated.