Application and drug of tetrahydrobiopterin
A technology of tetrahydrobiopterin and drugs, applied in the direction of drug combination, respiratory diseases, etc., to achieve the effect of enhancing anti-oxidation
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Embodiment 1
[0035] The oxidative inactivation of BH4 can be caused after the ionizing radiation of embodiment 1
[0036] The BH4 powder was dissolved in double-distilled water with a final concentration of 1 μg / ml. The aqueous solution was divided into two groups, and after 0Gy and 10Gy X-ray irradiation, the ultraviolet absorption spectrum was detected. BH4 has three characteristic absorption peaks at 230nm, 280nm and 330nm respectively. The spectrum of BH4 was detected immediately after 10Gy of X-ray irradiation, and its spectrum did not change significantly. With the increase of time after irradiation, the characteristic absorption peaks at 230nm, 280nm and 330nm of BH4 decreased gradually.
[0037] Such as figure 1 (attachment1 figure 1 ), the results showed that ionizing radiation itself did not cause changes in the structure of BH4, but the structure of BH4 changed with the increase of time after irradiation.
Embodiment 2
[0038] Example 2 Anti-radiation effect test of BH4 on lung cells
[0039] 1. BH4 reverses the reduction of NO caused by ionizing radiation
[0040] Human embryonic lung fibroblasts in the exponential growth phase were inoculated in 96-well plates and cultured for 12 hours. PBS or 12.5 μmol / L BH4 were added 24 hours before irradiation. Each group had 4 parallel samples, and the experiment was repeated 3 times. Afterwards, an absorbed dose of 20Gy X-rays was irradiated. Aspirate the medium 24 hours after exposure, add 100 μL of diluted DAF-FMDA to each well (dilute DAF-FMDA with DAF-FMDA diluent according to 1:1000, so that the final concentration is 5 μmol / L), and incubate in the incubator 20min. Wash three times with PBS to fully remove DAF-FMDA that did not enter the cells. Detection by microplate reader.
[0041] Such as figure 2 (P<0.05), the results showed that the BH4 treatment group had no effect on the NO content of the 0Gy group, but could significantly reverse ...
Embodiment 3
[0048] Example 3 Anti-radiation effect test of BH4 on mouse lung tissue
[0049] Male C57 mice aged 6 to 8 weeks were purchased from Shanghai Slack Experimental Animal Co., Ltd., and each mouse was anesthetized by intraperitoneal injection of 4% chloral hydrate according to 1% of its body weight, and small animal positioning was performed using a small animal precision irradiation device Afterwards, the right lung was precisely irradiated with an aperture of 5mm, so that 100% of the volume reached the prescribed dose of 20Gy, and a mouse radiation-induced lung injury model was established, and then the mice were randomly divided into 2 groups (5 in each group):
[0050] 1) Give mice a tail vein injection of 100 μL of BH4 aqueous solution at a concentration (1 mg / kg); 2) Give mice a tail vein injection of 100 μL of PBS.
[0051] After 7 days of irradiation, C57 mice were sacrificed by cervical dislocation. The right lung tissue was taken, rinsed repeatedly with normal saline, ...
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