| Viral Myocarditis(VM) could be harm to the health of public severely, but no patent medicine for(VM) was applied in clinical therapeutics nowadays . Therapeutic drugs mainly were divided into several kinds, which included immunosuppressant, chemical agents, active components of Chinese materia medica. Among of them the researches on active components of Chinese materia medica became the focus of study therapeutic methods recent years.The antiviral activity of KBD was studied by establishing experimental model in vitro and in vivo, and groups included pharmacal groups, positive drug control groups, control groups and viral groups.Experiment included two parts which were the inhibition of KBD on cells affection and multiplication of CoxB3 virus in cells. The first part was studied through observing the CPE and MTT(OD570nm) of the groups. And the process of another experiment was that the cells which were infected by virus were broken and centrifugated, collected supernatant, and measure virus tite.Experimental models is established (in vivo ) by intraperitoneal injection of Balb/c mice. Administration (KBD) is used to cure the sick mice, measured and analyzed Physiological and biochemical index were determined and analyzed during the whole experiment.1. The exosomatic experiments 1.1 The toxicity test of KBD in vitroKBD was prepared concentration gradient of 10 groups from600μg?ml-1 to 18.75μg?ml-1 and were added to the cell culture board which covered with monolayer cells, determined Maximal non-cytotoxicity concentrations(TD0) and 50% cytotoxicity concentrations(TD50).The Vero cells and myocardial cells′TD0 all is 75.00μg?ml-1 and The TD50 respectively is 187.49μg?ml-1 and 161.00μg?ml-1。1.2 The test of virulenceCoxB3 virus is attenuated 6 concentrations from 10-1 to 10-6 and added to the cell culture board which covered with monolayer cells. Virulence is determined and calculated by the Reed-Muench TCID50 is 10-2·ml.1.3 The experiment of affection inhibition on cellsKBD was prepared 5 groups from 75μg?ml-1 to 18.75μg?ml-1 and added to the cell culture board which covered with monolayer cells. GH and OL groups was used as positive drug control groups, control groups and viral groups is established in experiment. Data for t test could be determined by MTT (OD570nm) method. Compared to positive drug control groups the action of inhibition of drug group is significant t deviation. (P <0.05).1.4 The inhibition experiment on cells infested by CoxB3 virusThe cell which infested by CoxB3 virus were cured by KBD at different time. The cells which were infected by virus were broken and centrifugated, collected supernatant, and measure virus tite. The cells of viral group whose CPE achieve ++++ were cleavaged and centrifugated, and collected supernatant for measuring tite. Compared the lgTCID50 between the pharmacal groups and the viral group KBD was considered effective to virus, and difference is larger than one. Compared to viral control groups the action of inhibition of drug group is significant t deviation. (P <0.05).2. The endosomatic experiment2.1 The metered test of virulence in vivoCoxB3 virus was attenuated into 6 concentrations from 10-1 to 10-6 and injected intraperitoneally before the experiment′s seven day of Balb/c mice, calculated the death rate of mice and The LD50 of CoxB3 virus is 10-1.2.2 The effection of KBD on the usually states,weights and suviral rates of the miceThe activities and some kinds of index of mice was decreased after establishing model. Compared survival rate of two groups mice, results showed significant deviation (P <0.05).2.3 The experiment of inducing IFN on splenic cells and the experiment of IFN,s dynamic changesSpleens of mice were taken in the bioclean condition 7 days after being treating with KBD. Splenic cells and Con A were cultured in the cell culture board. Supernates were collected and centrifugated . The standard substance of IFN was made 7 concentrations. The regression equation of data was shown: Y=0.0025X+0.2847.Calculate the IFN valence ,then compare to the control group. The KBD group showed significant deviation (P <0.05). Its showed that KBD can enhancement Splenic cells′IFN excrete.2.4 The inhibitive action of KBD on releasing of the enzymes from myocardial cellsThe blood of mice is taken and supernated for measurement at fifth day and tenth day of administration duration in the bioclean condition. As was shown from data results , LDH, AST, CK and HBDH of blood-serum increased, and compare to viral group, high and mid doses can significantly reduced the index, all present significant deviation(P <0.01).2.5 The inhibition of KBD on multiplication of CoxB3 virus in vivoThe mice are executed and obtain hearts in the bioclean condition after being treating, were grinded in the grinder, and cleavaged and centrifugated. The supernates was added to the cell culture board which covered with Vero cells, observed after 24h, then calculated the lgTCID50. Compared the lgTCID50 between the pharmacal groups and the viral group KBD was considered effective to virus, if difference is larger than one.Pharmacal groups are effective that proved by the experiment. Significant deviation was shown between the pharmacal groups and the viral group (P <0.05).2.6 The inhibition of KBD on pathological change of myocardial tissueThe mice are executed and obtain hearts in the bioclean condition after being treating. The hearts were fixed by formalin and made pathological sections. KBD could reduce the affection which caused by virus, and effectives of it were better than that of positive drug.
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