Compound for blocking out interaction between uPA and uPAR, and application
A technology of compounds and fragments, applied in the direction of DNA/RNA fragments, hybrid peptides, drug combinations, etc., can solve the problem of preventing or treating atherosclerosis without ATF and anti-ATF antibodies
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Embodiment 1
[0110] Embodiment 1: Preparation of recombinant human ATF (rhATF):
[0111] According to the literature (Wang Jing et al., Preparation and Activity Measurement of Recombinant Human Prourokinase Amino-terminal Fragment, Journal of Nanjing University, January 2004, Volume 40, No. 1, p64-74), human umbilical cord vein endothelial cells The total RNA was purified from (HUVEC), and the human ATF gene was obtained by RT-PCR. The ATF gene was cloned into E. coli expression vector PET-29a (+), and transformed into E. coli BL21 (DE3) host bacteria. The rhATF expressed in the fermentation broth was denatured, and then purified by CM column and Superdex G-75 column, and the purity of the obtained sample was greater than 95%. Obtaining rhATF can prevent the binding of uPA and uPAR.
Embodiment 2
[0112] Example 2 Atherosclerotic lesion formation in rabbit aortic balloon injury model:
[0113] Take healthy male New Zealand white rabbits (2.8-3.5Kg) to carry out the following experiments: inject pentobarbital sodium (30mg / kg) into the ear vein to anesthetize the rabbit, fix it in the supine position, cut the right femoral artery, insert a 4F Fogarty catheter 20cm To the abdominal aorta, inject 0.85ml of normal saline into the balloon to fill, gently pull the balloon to fill up to 10cm, repeat four times to ensure that the endothelium is completely damaged, take out the catheter, suture the wound, and apply 200,000 IU of penicillin saline to the wound to prevent infection.
[0114] After the operation, the rabbits were immediately fed with high-fat feed (HFD, 1% cholesterol, 5% lard and 7.5% egg yolk powder were added to the common feed) for 7, 10, and 14 days. After the animals were sacrificed, the following procedure was used to observe the formation of atherosclerotic ...
Embodiment 3
[0116] Example 3 Intravenous injection of rhATF prevents atherosclerotic lesion formation in damaged arteries of rabbits:
[0117] After arterial balloon injury, the experimental group received 1 mg rhATF intravenously, once a day, for seven days; the control group received PBS intravenously. Feed a high-fat diet (HFD) during this period. The rabbits were sacrificed on the seventh day after the arterial balloon injury, and the formation of atherosclerotic lesions in the injured aorta was observed.
[0118] The results of this experiment are shown in Fig. 6 and Table 1. Compared with the control group injected with PBS, the atherosclerotic damage in the injured aorta of the experimental group injected with rhATF was reduced by about 50%.
[0119] Percentage of damaged area (%)
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