Application of baicalin in preparation of medicine for treating Marek's disease of chicken
A technology of chicken Marek's disease and baicalin, which is applied in the fields of medicine and antiviral applications, can solve the problems of enhancement and aggravation of symptoms, and achieve the effects of inhibiting replication, reducing cell lesions, and significant infection effects
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Embodiment 1
[0025] Effects of baicalin on the replication of chicken Marek's virus in CEFs.
[0026] figure 1 It is the CCK-8 result graph showing the effect of baicalin on the activity of chicken embryo fibroblasts (CEF); select baicalin at a safe concentration of 20 μg / mL and 2 μg / mL to pretreat CEF in a 12-well plate, and inoculate 200 PFU of MDV virus after 2 hours After 4 hours of adsorption in a 37°C incubator, the maintenance solution containing the corresponding concentration of baicalin was changed until the cells were harvested. 96 hours after inoculation, some cells were digested and used for counting viral plaques, and RNA was extracted from some cells for real-time PCR to detect the expression levels of Meq and gB genes. Some cells were lysed with RIPA buffer for western blot assay. The remaining virus infected CEF cells for indirect immunofluorescence detection of virus proliferation in the cells. It was found that, compared with the control group, when the concentration ...
Embodiment 2
[0028] Time dependence of baicalin on chicken Marek's virus inhibition in cells.
[0029] The virus-infected CEF cells were harvested at 24, 48, 72, 96, and 120 hours after virus infection, respectively, and used for virus plaque counting, qRT-PCR and indirect immunofluorescence tests. Real-time PCR results showed that the expression levels of Meq and gB genes in CEFs infected with MDV RB-1B and without drugs showed a significant increasing trend over time, while the expression levels of Meq and gB genes in CEFs infected with MDV RB-1B and added drugs The increase in expression was not obvious ( image 3 Middle A, B). Viral plaque counts and IFA results were also consistent with Real-time PCR results ( image 3 Middle C, D). The results showed that 20μg / mL baicalin had a significant effect on inhibiting MDV replication within 120 hours after infection.
Embodiment 3
[0031] Baicalin directly affects the infectivity of MDV virus
[0032] Suspend 200PFU virus with DMEM maintenance solution containing normal 0.5% serum and maintenance solution containing 20 μg / mL drug respectively, and incubate at 37° C. in a 5% CO2 incubator for 1.5 hours. After centrifugation to remove the supernatant, the cells were resuspended and seeded in CEF in a 12-well plate. After 4 hours of adsorption, fresh 0.5% serum maintenance solution was replaced until the virus-infected cells were harvested after 96 hours. Cells were collected for plaque counting and real-time PCR to detect virus proliferation.
[0033] The results showed that the viral gene expression in the drug treatment group ( Figure 4 Middle A) and virus proliferation ( Figure 4 Middle B, C) were significantly lower than the normal virus suspension group. This result indicated that 20 μg / mL baicalin could directly inhibit the infectivity of MDV.
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