| Objective: To explore whether curcumin works on atherosclerosis and to clarify its mechanism by detecting inflammatory factors associated with atherosclerotic inflammation and lipid metabolism,blood-fat levels,expression levels of proteins and genes in PI3K/Akt signaling pathway.Methods: 1.Animal experiment: 72 healthy rats were randomly divided into 6 groups: blank control group(group K),model control group(group M),simvastatin positive group(group Y),high,medium and low doses of curcumin groups(group G,Z and D).After a week of adaptive feeding,the other 5 groups of rats were given high-fat diet(80.8% basic diet +10% lard +5% white sugar +3.5% cholesterol +0.5% sodium cholate +0.2% propylthiouracil),with free access to water,except the blank control group.Each rat in the 5 groups was intraperitoneally injected with 600,000 IU/kg vitamin D3 at the beginning of the modeling,and 100,000 IU/kg vitamin D3 at weeks 3,6,9 and 12 after the modeling.Replicate the rat AS model with a modeling cycle of 12 weeks.2.Test indexes :(1)Blood was collected from abdominal aorta after the last dose and anesthesia with 10% chloral hydrate.Then serum TG,TC,LDL,TNF-α and il-6 were measured by ELISA after centrifugation at 4℃ and 3000 rpm for 10 min.(2)Open the chest cavity,and take the complete aorta out,then cut in half.One half was fixed with paraformaldehyde,dehydrated with ethanol gradient and embedded in paraffin,sliced up and stained with HE.Finally,observe the pathological changes of the aorta tissue.(3)The expression of PI3 K and Akt in the remaining aortic tissues was detected by western blotting.(4)The expressions of PI3 K and Akt mRNA were detected by RT-PCR.Results:(1)The results of one-way analysis of variance showed that the levels of IL-6 and TNF-αin the 6 groups were statistically significant(F=56.40,65.40,P<0.05),Q test results showed that except for no statistically significant difference in IL-6 between group M and group D,there were statistically significant differences between any two groups(P<0.05);the levels of TC,TG and LDL in the 6 groups were statistically significant(F=39.66-54.60,P<0.05),Q test results showed that except for no statistically significant difference in TC between group M and group D,group Y and group G,group G and group Z,in TG between group M and group D,group G and group Z,and in LDL between group K and group Y,group G and group Z,there were statistically significant differences between any two groups(P<0.05).(2)Pathological morphologic changes of aorta: HE staining showed that,compared with group K,a large number of inflammatory cells,foam cells and significantly proliferating smooth muscle cells were observed in the aortic tissues of group M,with scattered calcification.Inflammatory cells and foam cells were significantly reduced in group Y.Inflammatory cells,foam cells and proliferating smooth muscle cells were still observed in group G,Z and D,but the above pathological changes were dramatically alleviated compared with group M.(3)Western blot detection for protein expression: The expression of PI3 K and Akt in aortic tissues at protein levels in the 6 groups were statistically significant(F=878.68、297.83,P<0.05),with significant differences between any two groups(P<0.05).The expression of PI3 K and Akt in the model control group was significantly higher than that in the blank control group(P<0.05).Compared with the model control group,PI3 K and Akt protein expressions in simvastatin positive group,high and medium dose of curcumin groups were significantly reduced(P<0.05).(4)Detection of mRNA expression by RT-PCR :The expression of PI3 K and Akt in aortic tissues at mRNA in the 6 groups were statistically significant(F=2 930.20、2 874.12,P<0.05),with significant differences between any two groups(P<0.05).The expressions of PI3 K and Akt mRNA detected by RT-PCR were significantly higher in the model control group than that in the blank control group(P<0.05).Compared with themodel control group,The expressions of PI3 K and Akt mRNA were significantly reduced in simvastatin positive group,high and medium dose of curcumin groups(P<0.05).Conclusions: 1.In the AS rat model,curcumin could significantly inhibit the production of inflammatory cytokines IL-6 and TNF-αand reduce the levels of TC,TG and LDL,alleviate aortic inflammatory infiltration and accumulation of lipids,showing a good anti-AS effect.2.Curcumin can inhibit the expression of PI3 K,Akt genes and their proteins,regulate the apoptosis of inflammatory factors through this pathway,improve the metabolism of lipid,accelerate the elimination of autophagy of LDL-based lipid deposited in the aorta,and then alleviate the symptoms of atherosclerosis,which provides a theoretical basis for the research and development of as therapeutic drugs. |
