Application of 4-isothiourea butyronitrile hydrochloride in treatment of mycobacterial infection
A technology of isothiourea-butyronitrile hydrochloride and mycobacteria, which is applied in the field of medicine, can solve the problems of rifampicin treatment fatigue, achieve the effect of solving the problem of drug resistance and inhibiting survival
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Embodiment 1
[0032] In this embodiment, the damage of 4-isothioureidobutyronitrile hydrochloride to cells is determined by in vitro experiments, so as to delineate the safe range of medication. The specific operations are as follows:
[0033] 1. Experimental materials
[0034] Cells: RAW264.7 mouse mononuclear macrophage leukemia cells;
[0035] Reagents: dimethyl sulfoxide (DMSO), DMEM high glucose cell culture medium, fetal bovine serum, phosphate buffer saline, 4-isothioureidobutyronitrile hydrochloride powder.
[0036] 2. Experimental steps
[0037] Fetal calf serum was added to the cell culture medium, and RAW264.7 cells were seeded into 96-well cell culture plates, 1*10 per well 4 After the cells adhere to the wall, add 4-isothiourea-butyronitrile hydrochloride solution (prepared with DMSO). This experiment sets up five experimental groups, which are respectively 4-isothiourea-butyronitrile hydrochloride The concentration is 10, 20, 30, 40, 50umol / L groups, and the control group w...
Embodiment 2
[0044] The present embodiment confirms by in vitro experiments that 4-isothioureidobutyronitrile hydrochloride and its combination with rifampicin can effectively inhibit the survival of mycobacteria in macrophages. The specific operations are as follows:
[0045] 1. Experimental materials
[0046] Cells: RAW264.7 mouse mononuclear macrophage leukemia cells;
[0047] Bacteria: Mycobacterium smegmatis;
[0048] Reagents: dimethylsulfoxide (DMSO), DMEM high-glucose cell culture medium, fetal bovine serum, phosphate buffer (for washing cells before digestion and preparing cell lysate), Triton X-100 (for preparing 1 % cell lysate), Middlebrook7H9 broth base, agar powder, 4-isothiourea butyronitrile hydrochloride powder, rifampicin powder.
[0049] 2. Experimental steps
[0050] Fetal calf serum was added to the cell culture medium, and RAW264.7 cells were seeded into cell culture well plates, 1*10 per well 6 cells; after the cells adhered to the wall, add Mycobacterium smegmat...
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