Application of peroxy-ergosterol in drug resisting influenza virus infection and preparation of peroxy-ergosterol
A technology of peroxyergosterol and drugs, which is applied in the field of medicine, can solve problems such as undiscovered peroxyergosterol, and achieve the effect of inhibiting abnormal immune inflammation and mediating cell apoptosis
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Embodiment 1
[0030] Embodiment 1 prepares Nanbanlangen peroxyergosterol compound
[0031] Step S1: Extract
[0032] Cut 3Kg of Radix radix Radix into pieces, extract three times with 80% (volume) ethanol aqueous solution under heating and reflux, each time the volume of ethanol is 30L, 24L, 18L, to obtain ethanol extract, combine the ethanol extract, heat and depressurize recovery Ethanol to obtain the total extract extract.
[0033] Step S2: Extraction
[0034] Mix the obtained extract with water, heat and stir, extract the resulting mixture with 1 L of chloroform three times to obtain a water layer and a chloroform layer, and then use 1 L of ethyl acetate to extract the obtained water layer three times to obtain an ethyl acetate layer and a water layer . The chloroform layer and the ethyl acetate layer were respectively combined, and the solvent was recovered by heating and reducing pressure to obtain the chloroform layer extract and the ethyl acetate layer extract.
[0035] Step S3:...
Embodiment 2
[0045] Example Diperoxyergosterol Effects of Influenza Virus Infection Induced Host Expression of Pattern Recognition Receptor RIG-I
[0046] 1. Experimental materials:
[0047] 1.1 Drug test samples
[0048] Peroxyergosterol (prepared in Example 1) has a purity of 95% as detected by HPLC.
[0049] 1.2 Cells and strains
[0050] A549 cells were purchased from ATCC and kept in our laboratory; Influenza A H1N1 influenza virus PR8 strain (A / PR / 8 / 34, H1N1) was kept in our laboratory
[0051] 1.3 Reagents
[0052] Reverse transcription reaction kit (purchased from Takara company; catalog number: RR036A); real-time quantitative PCR reaction kit (purchased from Takara company; catalog number: RR390A)
[0053] 2. Experimental method
[0054] Including the following steps:
[0055] 1) A549 cells were mixed with 5X10 6 The cell density was planted in a 6-well plate, and placed in a temperature of 37 ° C, 5% CO 2 in the incubator;
[0056] 2) After culturing the cells overnight,...
Embodiment 3
[0079] Example 3 Effect of ergosterol peroxygen on the expression of inflammatory genes in the host induced by influenza virus infection
[0080] 1. Experimental materials:
[0081] 1.1 Drug test samples
[0082] Peroxyergosterol (prepared in Example 1).
[0083] 1.2 Cells and strains
[0084] A549 cells were purchased from ATCC and kept in our laboratory; Influenza A H1N1 influenza virus PR8 strain (A / PR / 8 / 34, H1N1) was kept in our laboratory.
[0085] 1.3 Reagents
[0086] Reverse transcription reaction kit (purchased from Takara company; catalog number: RR036A); real-time quantitative PCR reaction kit (purchased from Takara company; catalog number: RR390A)
[0087] 2. Experimental method,
[0088] Including the following steps:
[0089] 1) A549 cells were mixed with 5X10 6 The cell density was seeded into a 6-well plate, placed at a temperature of 37 ° C, 5% CO 2 in the incubator;
[0090] 2) After culturing the cells overnight, discard the original medium and add PB...
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