Application of berberine in medicine for treating or preventing influenza virus
A technology for preventing influenza and berberine, applied in the field of medicine, to achieve the effect of inhibiting inflammatory response, alleviating treatment pain and having less toxic and side effects
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Embodiment 1
[0057] Example 1: Evaluation of berberine anti-influenza virus activity
[0058] 1. Materials: cells, viruses and plasmids:
[0059] MDCK cells were provided by Shandong Institute of Medical Microbiology, Chinese Academy of Sciences; A549 cells, HeLa cells, virus H5N1 or H3N2 were purchased from China Collection Center for Typical Microorganisms; pVP-16-NP, pM-PB1, pM-PB2, pVP-16-NP, The pG5Luc plasmid was completed under the guidance of Professor Wu Jianguo of the Medical Virology Research Group of the State Key Laboratory of Virology, Wuhan University. pG5Luc is a reporter gene carrier with green fluorescent GFP, and the plasmids pVP-16-NP, pM-PB1, pM-PB2, pVP -16-NP are different proteins of influenza virus connected to the vectors pVP and pM, and the relevant details can be found in the papers published by our laboratory——Mukhtar, M.M., Li, S., Li, W., Wan, T. , Mu, Y., Wei, W., Kang, L., Rasool, S.T., Xiao, Y., Zhu, Y., and Wu, J. (2009).Single-chain intracellular antibo...
Embodiment 2
[0159] Example 2: The effect of berberine on prevention of virus infection, adsorption, invasion and replication of influenza virus
[0160] The cells paved into a single layer were treated in different ways: the drug was added to the cells in advance and incubated for 1 hour, the medium was changed, and the corresponding influenza virus was added; the drug was added to the virus for adsorption and invasion of the influenza virus, and the drug was added to the virus and incubated for 1 hour, and then Then add it to the cells; for the effect of the drug on the replication of the influenza virus, the virus is first used to infect the cells for 1 hour, and then the drug is added. The drug concentration used in all experiments was 100ug / ml, and the virus infection amount was the virus liquid of 1M.O.I. Part of the supernatant was collected at 24, 36, and 48 hours respectively, and the antiviral effect of the drug was measured by HA.
[0161] The results are shown in Figure 9. Ber...
Embodiment 3
[0162] Example 3: Two-hybrid experiments in mammals to explore the effects of drugs on some enzymes in the replication process
[0163] 1. Cell transfection
[0164] The plasmids are pM-PB1, pM-PB2, pVP-16-NP, pG5Luc, respectively. In this test group, no plasmids were added to the negative control, pM-PB1 (or pM-PB2), pVP-16-NP, and pG5Luc were added to the positive control, and pM-PB1 (or pM-PB2), pVP- 16-NP, pG5Luc three plasmids and various drugs.
[0165] Sofast TM lipofection
[0166] The following transfection operation uses a 24-well plate as an example. The whole process of transfection must be strictly aseptic.
[0167] 1. Seed the cells to be transfected in a 24-well plate one day before transfection and culture at 37°C. The cell density before transfection was 40-60%.
[0168] 2. Prepare the following solutions
[0169] ① Dilute 1.5μl Sofast TM Reagents in 30 μl serum-free, antibiotic-free DMEM medium. After mixing, let stand at room temperature for 3 minu...
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