Medicine for treating flap transplantation injury and application thereof
A transplant injury and drug technology, applied in the field of medicine, can solve the problems of amplifying inflammatory response, cell damage, aggravating organ damage and apoptosis, etc., to improve necrosis, improve treatment effect, and reduce oxidative damage.
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Embodiment 1
[0019] Grouping of Study Animals
[0020] The animals participating in this study were male Wistar rats aged 12-14 weeks, weighing 280-330 grams (g), a total of 60 were purchased from Slac Experimental Animals (Shanghai, China).
[0021] In the case of sufficient supply of standard laboratory food and tap water, put them in SPF room, light-dark cycle 12h, 18-25°C, relative humidity 40-70%, and establish I / R model for rats. Wistar rats were divided into six experimental groups with 10 rats in each group:
[0022] Sham-operated group: (1) The superficial abdominal wall arteries and veins at the pedicle were separated after the flap design was cut; (2) The flap was sutured in situ without ischemia-reperfusion treatment; (3) There was no special treatment after operation.
[0023] Remifentanil-H group: Remifentanil (10 μg / kg) was pretreated on the basis of the sham operation group.
[0024] Group I / R: The flap design and cutting method were the same as those in the sham operatio...
Embodiment 2
[0029] Aortic endothelial cell harvesting and cell culture
[0030] To study the effect of remifentanil at the cellular level, aortic endothelial cells were isolated from epigastric flaps of 20 Wistar rats aged 12-14 weeks and weighing 280-330 g.
[0031] All rats received an adequate supply of standard laboratory chow and tap water, and were housed in an SPF chamber with a 12-h light-dark cycle, 18-25°C, and 40-70% relative humidity.
[0032] Isolation of abdominal aortic endothelial cells: After anesthetized with pentobarbital sodium salt, cut the abdominal aorta according to the abdominal tangent, and perfuse the left ventricle with 1 mL of fetal bovine serum containing heparin (1000 U / mL); the aorta was obtained from a rat After taking it out, incubate in DMEM, add 20% fetal bovine serum containing heparin (1000U / mL); after the proximal end of the aorta is ligated, wash the ligated section with serum-free DMEM medium, and incubate the other side at 37°C with II Type colla...
Embodiment 3
[0034] cell treatment
[0035] The I / R injury model of aortic endothelial cells treated with hypoxia-reoxygenation was established, and the aortic endothelial cells were divided into the following groups:
[0036] Control group: cells were cultured at 37°C in DMEM medium containing 15% fetal bovine serum;
[0037] Hypoxia-reoxygenation group: cells were incubated at 37°C, 5% CO2, 3% O2, 92% N2 in DMEM medium for 12h, and then incubated at 37°C, 5% CO2, 95% O2 in serum-free DMEM medium 4h, hypoxia-reoxygenation treatment;
[0038] Hypoxia-reoxygenation + remifentanil group: before the hypoxia-reoxygenation treatment, the cells were treated with 0.1ng / mL remifentanil medium;
[0039] Hypoxia-reoxygenation + remifentanil + anisomycin (Anisomycin) group: before the hypoxia-reoxygenation treatment, the culture medium was treated with 1 ng / mL remifentanil and 10 ng / mL anisomycin.
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