Application of wogonin in preparing medicine for resisting herpes simplex virus
A technology of herpes simplex virus and wogonin, applied in antiviral agents, pharmaceutical formulas, medical preparations containing active ingredients, etc., can solve the problems of undisclosed anti-infection effects of wogonin, and achieve strong inhibition of herpes simplex virus Infectious activity, mitigation of HSV drug resistance, and low cytotoxicity
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Embodiment 1
[0029] Example 1: Wogonin inhibits the pathological effect of HEC-1-A cells caused by herpes simplex virus
[0030] HEC-1-A cells are derived from human endocervical adenocarcinoma. Previous studies have shown that they can support the infection and replication of HSV-1 and HSV-2 in their cells. Like Vero cells in HSV virology research, it is also a good cell model for studying the virus (Qiu M, et al. Pyrrolidine dithiocarbamate inhibits herpes simplex virus type 1 and 2 replication and its activity maybe mediated through dysregulation of ubiquitin-proteasome system. Journal of Virology , 2013, 87(15): 8675-8686.).
[0031] The HEC-1-A cells were divided into 4 groups, the first group was the blank control group, normal cultured HEC-1-A cells; the second group was the virus control group (HSV-2), added HSV-2 (G) HEC-1-A cells cultured in virus solution (moi=1); Group 3 is HEC-1-A cells incubated with Scutellaria baicalensis alone (Wog.), cultured with wogonin (final concentr...
Embodiment 2
[0034] Embodiment 2: Wogonin inhibits the replication test of herpes simplex virus on HEC-1-A cells
[0035] HEC-1-A cells were seeded on a 96-well plate at an appropriate density. After the cells were fused the next day, firstly, the cells were pre-incubated with gradiently diluted wogonin solutions of different concentrations, and then HSV-1 (HF) was added respectively. Or HSV-2 (G) (moi=1) to infect the cells. After culturing for 24 hrs, 200 μL of new medium was added to each well, and the cells in the 96-well plate were transferred to minus 80°C and 37°C for three times of freezing and thawing to release virus particles. The virus solution was diluted 5-fold, and 50 μL of the diluted solution was added to a 96-well plate with a monolayer of HEC-1-A cells, and the number of plaques was counted after 48 hr of incubation to calculate the virus titer.
[0036] The results of the plaque counting experiment were as follows: figure 2 It was shown that wogonin had inhibitory ef...
Embodiment 3
[0037] Example 3: Wogonin inhibits the replication test of herpes simplex disease at the RNA level
[0038] HEC-1-A cells were seeded in 24-well cell culture plates. After the cells were confluent, wogonin solutions of different concentrations were added in gradient dilutions, incubated at 37°C for 30 min, and then HSV-1 (HF) or HSV were added respectively. -2 (G), infection titer moi=1. After 24 hrs, discard the medium, add 200 μL Trizol to lyse the cells, and extract the total cellular RNA according to the steps described in the manual.
[0039] Using the TOYOBO reverse transcription kit instructions, the first strand of cDNA was synthesized and then qPCR was detected, and the HSV-1 gD gene (GenBank ID: 2703444), HSV-2 gD gene (GenBank ID: 1487358) and internal reference were detected by SYBR Green method Gene GAPDH amplification. The reaction system was 20 μL, and the reaction conditions were pre-denaturation at 95°C for 10 min; 95°C for 15 s, 60°C for 15 s, 72°C for 45 s,...
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