| 1. To study the therapeutic effects of Scutellaria baicalensis (SB) on coxsakievirus B3m (CVB3m) in vitro, and to explore the antiviral mechanisms, and to observe the influence on cell activity by SB.2. To exam the pharmacokinetic parameters of SB in rat and test the distribution of SB in rat's different organs.3. To explain how SB antivirus towards CVB3m in vivo and the target effectson myocardiac immuno-injury (apoptosis and CMC) caused by CVB3minfection. Methods 1. We used microcell culture method and dye absorbed methods to recordcytopathic effects (CPE) and cell activities. From this work, we candetermine the TD0, TD50 of SB on the cultured cells and the restrainedindexes or percents of SB towards CVB3m.2. The rats was dealed with 40mg/kg of SB, the urine, heart, liver and kedneyof rats were collected at different times, and then HPLC was applied to detect the cotent of SB in the different specimens.3. BALB/c mice were inoculated intraperitoneally with CVB3m and batchsacrificed late. On the base of the models, following experiments were carried.3. 1 The CVB3m RNA were detected by reverse transcription-polymerase chain reaction (RT-PCR) , the RT-PCR product were coloned in plasmid to test the DNA sequence.3. 2 Mice were randomly dividedinto 4 groups, the model group, the positive control group (treated with Ribavirin), the SB group and the normal control group. Body weight of mice, food and water that mice ate, VMC symtops and death of mice were observed at the 3th, 5th, 7th, 10th and 14th day. Weight, macro-, histopathologic and ultramicro changes of heart were were observed and measured by morphometric quantification, and the data were treated statistically.3. 3 Immunohistochemical observations, including apoptosis by TDT/Dutp Nick End labeling (TUNEL), were performed on mice hearts, and the results weredetected by morphometric analysis system. 3. 4 The iNOS mRNA and CVB3m RNA were examed on mice hearts by RT-PCR methodto study the relation between them in VMC. 3. 5 The apoptosis related gemeno of Bcl-2 and Fas mRNA were examed on micehearts by RT-PCR method to explore the regulating mechanism of SB onmycardic cell apoptosis. Results1. After camparing the different links of SB on CVB3m and cultured cells, the results indicated SB had the effects of protecting cultured cells from CVB3m infection and killing CVB3m indirectly in vitro.2. The HPLC mesurement data and analysis results showed the metabolic procedure of SB was similar as two-room model, and SB was highly distributed in heart and liver of rats.3. Animal experiments3.1 The DNA sequence was similar as CVB3m cDNA by 99.5%, and the histopathologic change of model mice was similar to VMC mice.3. 2 The myocarditis was detected from all CVB3m_infected mice and was most severe at 5-10 days, the total mortality was 70%. Body weight of mice, food and water that mice ate decreased after infection in model group,weight and the index of heart increased. In the SB group, the rates of VMC and death were lower than the model and positive control groups. The macro-, histopathologic and ultramicro changes of mice heart of the SB group were better than the model group too.3.3 Immunoreactivities for iNOS and Fas increased considerably in myocardium after infection, and decreased gradually after 10-14 days, significant difference was found when compared with the normal group. Higher apoptosis in acute stage of infection, low apoptosis in chronic stage and no apoptosis in normal control group were found. In the SB group, myocardium apoptosis, iNOS and Fas expression were higher than the model group and the positive control group.3.4 iNOS mRNA expression is higher in CVBa, infected mice than that in the normal control group. iNOS mRNA expression have positive relationship with CVB3m RNA. In the SB group, iNOS mRNA expression was higher than the model group and the positive control group, while CVB3m RNA in mice heart was lower than those group...
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