| BackgroundArterial atherosclerosis(AS)is a disease with pathological damage of brain tissue as the main clinical manifestation.It has a high morbidity and mortality,and is one of the main diseases that threaten human health.)Is the initial stage of contractile stroke.The formation of plaque makes the diameter or occlusion of blood vessels.Plaque displacement is the main pathological change of atherosclerosis.The pathogenesis of atherosclerosis is very complicated.At present,the pathological features of atherosclerosis are mainly:lipid infiltration after vascular intimal injury,mononuclear macrophages,smooth muscle cell proliferation and foam cell formation.The most commonly used drugs for the treatment of atherosclerotic plaques are statins,but the effects of statins on the treatment of plaques are controversial and some patients cannot tolerate them.Therefore,the development of drugs and methods for the treatment of choroid-like sclerosis is also a current research hotspot.Time-restricted feeding(TRF)is a type of intermittent fasting(IF)that is more acceptable than traditional volume restrictions.Most of TRF's research on implanted animals and humans has focused on reducing high-fat diets.Some preliminary results from animal experiments have shown that TRF can delay the development of neurodegenerative diseases and improve cognitive function;it may also relieve sexual stroke Inflammatory reaction in mice,but the mechanism of TRF on atherosclerosis has been reported.In this study,the recombinant gene knockout mouse atherosclerosis model related inflammatory factors were started to explore the effect of TRF on atherosclerosis mechanism.ObjectiveBy establishing an ApoE-/-mouse atherosclerosis model to detect changes in aortic vascular lesions and inflammation-related factors in mice after time-restricted feeding interventions,to explore the effects of time-restricted feeding on ApoE-/-mouse atherosclerotic aortic disease The influence of tissue provides a theoretical basis for the treatment of atherosclerosis.MethodA total of 70 SPF grade 6-7-week-old male apolipoprotein E gene knockout(ApoE-/-)mice were used.After 1 week of adaptive feeding,they were fed with high-fat feed for 12 weeks,and then randomly selected Six of them were sacrificed and examined by HE staining of aortic tissues.Atherosclerotic plaques appeared in the aortic roots,and AS mouse models were successfully constructed.2.Grouping The remaining 64 AS mice were randomly divided into an experimental group,the time-restricted feeding group(TRF group)and a control group,the ad libitum(AL group),32 in each group,and the TRF group ate 8 daily Normal feed for 16 hours,fasting for 16 hours,free intake of water;the AL group ate regular feed 24 hours a day and free intake of water.Group feeding for 28 days.3.At 08:00,7th,14th,21st,and 28th morning at 8:00 AM,8 AS mice in the TRF group and AL group were randomly selected to measure body weight,Blood glucose and blood ketones were collected by tail-cutting method.Blood glucose and blood ketone body tester was used to measure and record the blood glucose and blood ketone levels.4.After 28 days of group feeding,blood was taken from the retro-orbital venous plexus with eyeball removal and blood sampling,and serum was taken after centrifugation.Colorimetric determination of total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C)in mouse serum was performed by microplate reader Change.5.After 28 days of group feeding,HE staining was used to observe the pathological changes,lesion degree and plaque area changes of the aortic root.6.Randomly select 8 AS mice in TRF group and AL group at 8:00 AM on the 3rd,7th,14th,and 28th day of group feeding,and remove the blood from the back of the orbit with eyeball removal The venous plexus was sacrificed after blood sampling,and the serum was collected after centrifugation.The serum levels of interleukin 10(IL-10)and tumor necrosis factor-?(TNF-?)were detected by ELISA.7.After 28 days of group feeding,the expression of nuclear factor-?B(NF-?B)positive cells in the aortic root tissue was detected by immunohistochemistry.Result1.After feeding the male ApoE gene knockout mice with high-fat diet for 12 weeks,obvious AS plaques formed in the mouse aorta.2.After time-limited feeding,the weight of the mice in the TRF group was slightly lower than that in the AL group,but the difference in body weight between the two groups was not statistically significant(P>0.05),indicating that the time-limited diet had no significant effect on the weight of ApoE-/-mice.3.The blood glucose of mice in TRF group was lower than that in AL group,the difference was statistically significant(P<0.01).4.The blood ketone body of the TRF group was significantly higher than that of the AL group,so differences are statistically significant(P<0.01).5.After 4 weeks of group feeding,compared with the AL group,the serum TC and LDL-C levels in the TRF group decreased,so the difference was statistically significant(P<0.05).6.After 4 weeks of group feeding,compared with the AL group,the plaque area of the aortic root in the TRF group decreased,lipid deposition,cholesterol crystals,and foam cells in the plaque were reduced,The difference was statistically significant(P<0.05).7.On the 3rd and 7th day of group feeding,there was no significant difference in IL-10 in the serum of the two groups of mice(p>0.05).On the 14th and 28th day of group feeding,the IL-10 of TRF group was more than AL The group increased and the difference was statistically significant(p<0.05).The TNF-? of TRF group was lower than that of AL group on the 3rd,7th,14th and 28th days of the experiment,the difference was statistically significant(p<0.05).8.After 4 weeks of group feeding,the positive expression of NF-?B p65 in AS plaque in the aortic root of TRF group decreased,compared with the AL group,the difference was statistically significant(P<0.05).Conclusion1.Time-restricted feeding can reduce the area of aortic plaque in the AS mouse model,lower blood sugar levels,increase blood ketone body levels,and reduce serum TC and LDL-C levels.2.Time-restricted feeding can reduce the levels of inflammation-related factors NF-?B and TNF-? in the AS mouse model and increase the level of IL-10,suggesting that time-limited feeding may delay the progression of atherosclerosis by inhibiting the NF-?B signaling pathway. |
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