| Objective:A model of hindlimb ischemia in diabetic rats was established to investigate the effects of curcumin nicotinate on the ischemic hindlimbs of diabetic rats by regulating Notch1 protein.Methods:Fifty-six male SD rats(200±20g)were divided into 5 groups according to the random number table method: normal group,diabetes group,diabetes+surgical group,diabetes+surgery+low-dose nicotinate curcumin group,diabetes+surgery+high-dose curcumin nicotinate group,12 mice in each group.The drug group was given intragastric administration of curcumin nicotinate 10 mg/kg and 100 mg/kg respectively from the 3 days before surgery to 28 days after surgery.The rest groups were given equal volumes of 0.5% sodium carboxymethyl cellulose each day.After diabatic rats are modeled and stabilized for 2 weeks,the hind limb ischemia models were constructed by surgical ligation and cutting off the common femoral artery.On the 4 weeks after surgery,the gastrocnemius muscles were harvested.HE staining was used to observe the muscle necrosis,angiogenesis and inflammatory cell infiltration.Western Blot was used to detect the expression of Notch1 and CD31 protein in the specimens and the expression of SOD was detected by spectrophotometry.Results: The skin temperature of hindlimbs in the diabetic+surgical group and the diabetic+surgical+drug group decreased,but there was no evidence of toe necrosis.Hindlimb ischemia in the diabetic+surgical group was relatively obvious,and there was a condition of trailing feet,and it was necessary to rely on the crawling of the hind limbs.Hindlimb ischemia in the diabetic+surgical+drug group was improved,and the movement was normal or slightly distorted.HE staining showed that the striated muscle cells in the diabetic+surgical group were disordered,the cells were swollen,the cytoplasm was dense,the nuclear chromatin margin or flocculation and other signs of apoptosis and necrosis.In the drug group,striated muscle cells were arranged neatly,showing more capillaries and a small amount of inflammatory cell infiltration.The expression of Notch1 and CD31 protein in the gastrocnemius muscle of the operative side was detected by Western blot.It was found that diabetes can cause abnormally high expression of Notch1 protein(p<0.01),and the expression of Notch1 protein in the diabetic+surgical group was higher(p<0.01).The expression of Notch1 protein in the diabetic+surgical+drug group was significantly lower than that in the diabetic+surgical group(p<0.01).Diabetes inhibited the expression of CD31 protein(p<0.01).The concentration of CD31 protein in diabetes+surgery group was further decreased(p<0.01),and the expression of CD31 protein was promoted in treatmentgroup(p<0.01).And the high-dose curcumin nicotinate group inhibited Notch1 and promoted CD31 protein expression significantly higher than the low-dose curcumin nicotinate group(p<0.05).Detection of SOD protein by spectrophotometry found that diabetes inhibited the production of SOD protein(p<0.01).Ligation of the femoral artery further reduced SOD protein production(p<0.01)and aggravated oxidative stress injury.The treatment group promoted SOD protein expression(p<0.01),but different doses of curcumin nicotinate did not cause differential expression of SOD(p>0.05).Conclusion:1.Curcumin nicotinate may protect the ischemic hind limbs of diabetic rats by promoting angiogenesis,and high doses of curcumin nicotinate stimulate angiogenesis effect is more obvious;2.The mechanism of curcumin nicotinic acid esters promoting ischemic hindlimb angiogenesis in diabetic rats may be related to the down-regulation of Notch1 protein expression to inhibit oxidative stress. |
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