Application of isoorientin in preparation of medicament for resisting respiratory syncytial virus
An anti-respiratory syncytial virus technology, applied in the field of medicine, can solve the problems of non-specific prevention and control measures, high toxicity and side effects, and high cost, and achieve strong anti-respiratory syncytial virus activity, novel mechanism of action, and good application prospects
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Embodiment 1
[0016] Example 1: Anti-respiratory syncytial virus activity of isoorientin in vitro
[0017] (1) Cells, viruses and experimental materials: Respiratory syncytial virus (RSV, Long strain), host cells are laryngeal carcinoma cells (Hep-2 cells); positive control drug is ribavirin (ribavirin); cells grow at a mass percentage concentration 10% fetal bovine serum (FBS) in the MEM medium; the maintenance solution is the MEM medium containing 1% FBS in mass percent concentration.
[0018] (2) Preparation of sample solution: the isoorientin sample (purchased from SIGMA company) and ribavirin sample were made into a solution with a concentration of 40mg / mL with dimethyl sulfoxide (DMSO), and then respectively mixed with maintenance solution. The samples were prepared into 400 μg / mL and 25 μg / mL sample solutions, which were used for cytotoxicity and antiviral activity experiments, respectively.
[0019] (3) measure the cytotoxicity of compound with tetrazolium salt (MTT) colorimetry: ...
Embodiment 2
[0027] Example 2: Determination of in vitro anti-respiratory syncytial virus activity of isoorientin and ribavirin in combination
[0028] (1) Cells, viruses and experimental materials: Respiratory syncytial virus (RSV, Long strain), host cells are laryngeal carcinoma cells (Hep-2 cells); positive control drug is ribavirin (ribavirin); cells grow at a mass percentage concentration 10% fetal calf serum (FBS) MEM medium; the maintenance solution is 1% FBS MEM medium.
[0029] (2) Preparation of sample solution: the isoorientin sample (purchased from SIGMA Company) and ribavirin sample were made into solutions with a concentration of 40mg / mL with dimethyl sulfoxide (DMSO), and the isoorientin samples were respectively mixed with maintenance solution. Orientin and ribavirin were prepared into 5.7 μg / mL and 3.0 μg / mL solutions, and then the two were mixed at a volume ratio of 1:1 to form a sample solution to be tested.
[0030] (3) The cytopathic effect reduction assay was used to...
Embodiment 3
[0033] Example 3: Anti-respiratory syncytial virus activity of isoorientin in vivo
[0034] (1) Animals: Kunming male or female mice (18-22 g).
[0035] (2) Animal experiment: the mouse RSV infection model was used to measure the anti-respiratory syncytial virus (anti-RSV) activity of isoorientin.
[0036] Select 60 BALB / C mice, half male and half female, and randomly divide them into 6 groups, namely high-dose isorientin group (40.0mg / (kg·d)), middle-dose group (20.0mg / (kg·d) )), low dose group (10.0mg / (kg·d)), ribavirin 10mg / (kg·d), virus positive and normal saline negative control group, 10 rats in each group. Intragastric administration, positive and negative control groups intragastric administration of normal saline. Three days later, nasal instillation of RSV suspension (TCID 50 for 10 -6.6 / ml), continue to administer by intragastric administration for 7 days, and observe the survival situation of mice. The mice were sacrificed 40 minutes after the last administra...
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