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Objective To elucidate activity of baicalin against human cytomegalovirus ( HCMV) in vitro, and explore its effect on apoptosis of human embryo lung fibroblasts ( HEL ) infected with HCMV. Methods CCK-8 kit was used to determine the maximum tolerated dose ( MTC ) of HEL cell to baicalin while the anti-HCMV median efficacious concentration ( EC50 ) of baicalin was determined by standard plaque reduction method. After treatment with baicalin of different concentrations for 24, 48, 72 and 96 h, cell apoptosis and pro-caspase-3 expression was detected by flow cytometry and Western blotting, respectively. Results The MTC of baicalin was 20.6 μg?mL-1; The EC50 of anti-HCMV of baicalin was 16.13 μg?mL-1;The apoptosis rate increased gradually in the groups with low and high multiplicity HCMV infection at early time, showing significant dose-dependent manner. While the ratio of apoptotic cells was going to decrease in high multiplicity infection group 96 h after the infection. The expression of pro-caspase-3 was significantly higher in high-dose baicalin treatment group than in the infection control group ( P<0.05) . Conclusion Baicalin has a direct anti-HCMV effect in vitro. One of the mechanisms might be related with it inhibiting cell apoptosis and antagonizing activation of pro-caspase-3.
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Objective To elucidate activity of baicalin against human cytomegalovirus ( HCMV) in vitro, and explore its effect on apoptosis of human embryo lung fibroblasts ( HEL ) infected with HCMV. Methods CCK-8 kit was used to determine the maximum tolerated dose ( MTC ) of HEL cell to baicalin while the anti-HCMV median efficacious concentration ( EC50 ) of baicalin was determined by standard plaque reduction method. After treatment with baicalin of different concentrations for 24, 48, 72 and 96 h, cell apoptosis and pro-caspase-3 expression was detected by flow cytometry and Western blotting, respectively. Results The MTC of baicalin was 20.6 μg?mL-1; The EC50 of anti-HCMV of baicalin was 16.13 μg?mL-1;The apoptosis rate increased gradually in the groups with low and high multiplicity HCMV infection at early time, showing significant dose-dependent manner. While the ratio of apoptotic cells was going to decrease in high multiplicity infection group 96 h after the infection. The expression of pro-caspase-3 was significantly higher in high-dose baicalin treatment group than in the infection control group ( P<0.05) . Conclusion Baicalin has a direct anti-HCMV effect in vitro. One of the mechanisms might be related with it inhibiting cell apoptosis and antagonizing activation of pro-caspase-3.
RESUMO
Objective To explore the correlation between the expression of IL-17A and the degree of spleen damage in acute mouse cytomegalovirus(MCMV) disseminated infection in vivo and to understand the mechanism about how IL-17A involved in the pathological damage of the spleen in MCMV infection.Methods An acute disseminated MCMV infection model was established in mice.BALB/c mice were randomly divided into two groups.Mice in group one were infected with MCMV Smith to establish disseminated infection.Mice in another group were sham-infected control.Three mice from each group were randomly chosen to be sacrificed on days 3,7,14 and 28 after the infection.Viral titers in spleen tissues were determined using a standard plaque assay.The expression of IL-17A mRNA and MCMV mRNA in the splenocytes were measured by RT-PCR.The expression of IL-17A in spleen tissues was observed by immunohistochemical staining.The pathology of the infected mice was assessed by histological examination of H&E stained spleen sections.Results Viral titers and MCMV mRNA in the spleen peaked on day 3,but quickly diminished on day 7.Virus was no longer detectable in the spleen on day 14 after the infection.The expression of IL-17A mRNA was significantly increased during the acute infection and reached the highest level on day 14,then decreased on day 28.It is significantly higher than that of the mock infection group.Immunohistochemistry assay also indicated that the expression of IL-17A in spleen tissue gradually increased to climax on day 14,then decreased on day 28.Accordingly,the pathological damages of spleen tissue in the infected mice deteriorated until day 14,then showed signs of recovery on day 28.The most severe pathological injury of spleen tissue and the highest expression of IL-17A appeared in the same period of time.Conclusion Our results showed a close correlation between IL-17A and the pathological damage in spleen.Thus,IL-17A may contribute to the spleen pathological damage during the acute disseminated MCMV infection.