Study On Different Kinds And Doses Of Whole Grains On Glucolipid Metabolism And Its Mechanism

Objective:(1)To provide a theoretical and experimental basis for selecting whole grains on the prevention and treatment of metabolic diseases,it need to compare the effects of different types of whole grains on IR and detect whether there are differences.(2)To clarify the dose-effect relationship of whole grains to improve IR,and to provide an experimental basis for the recommended intake levels of whole grains for the prevention and treatment of metabolic diseases and diet of T2 DM.(3)It is proposed that whole grains can improve the intestinal flora,regulate TLR4/NF-κB signal transduction pathway and regulate inflammatory factors,the whole grains can influence the possible pathway of glycolipid metabolism afterwards.To elucidate the possible molecular mechanisms that whole grains improve glycolipid metabolism and inflammatory levels,and provide a theoretical basis for the application of whole grains in the T2 DM.Methods:(1)Study on different kinds of whole grains on glucolipid metabolism and its mechanism.90 SD rats were randomly divided into 9 groups according to the level of HOMA-IR: the negative control group,the insulin resistance model group,refined rice control group,fine noodles control group,wheat group,corn group,millet group,barley group,oat group.Given diet for 8 weeks,and extract blood.At the end of the experiment,rats were sacrificed,and the rat organs such as the intestine and stool were saved.(1)Fasting plasma glucose(FPG),fasting plasma insulin(FIN),glycated hemoglobin(HbAlc),C-Reactive protein(CRP),Tumor necrosis factor-alpha(TNF-α),Interleukine-6(IL-6),Total cholesterol(TC),Triglyceride(TG),High-density Lipoprotein-cholesterol(HDL-C),Low-density Lipoprotein-cholesterol(LDL-C)were measured.(2)Gene expressions of TLR4,TLR5,TNF-α,NF-kB in the intestine tissue were determined by a real-time method.Protein expression of IKB-α,MyD88,TLR4,NFkB in the intestine tissue determined by western blot method.(3)DNA was extracted from the stool,and the 16 S rDNA high-throughput sequencing technology was used for sequencing,At last,analyze OTUs clustering and species classification.(2)Study on different dose of oats on glucolipid metabolism and its mechanism.Through the first phase of animal testing,oats were selected as the target grain.70 SD rats were randomly divided into 9 groups according to the level of HOMA-IR: the negative control group,the insulin resistance model group,refined rice and fine noodles control group,25% oats group,50% oats group,75% oats group,100% oats group.Given diet for 8 weeks,and extract blood.And the rat organs such as the liver and stool were saved.(1)Serum determination and intestinal flora detection methods are the same as in the previous chapter.(2)Gene expressions of rSREBP1 c,rChREBP,rTLR4,rTNF-a in the liver tissue were determined by a real-time method.Protein expression of IKB-α,MyD88,TLR4,NF-kB in the liver tissue determined by western blot method.Results:(1)Study on different kinds of whole grains on glucolipid metabolism and its mechanism.(1)Animal experimental results of different whole grains show that the level of HDL-C increase obviously(P<0.05)when comparing the level of TG,TC,LDL-C,FPG,FINS,HbAlc,HOMA-IR group with refined rice control group(fine noodles control group(P<0.05)).There is no significant difference in five grains(P>0.05),however,the effect of improving the blood sugar and blood lipids in the oat group is better than the other four grains,and the level of the rice and noodles group was comparable to that of the insulin resistance model group.Compared with the polished rice control group and the fine control group,barley,millet,and oatmeal could reduce the concentration of LPS(P<0.05),while there was no significant difference between wheat,corn group,and rice,noodle group(P>0.05).Five grains could reduce the concentration of hs-CRP and IL-6 in rats,which was lower than that in the polished rice group and the fine surface group(P<0.05).The IL-6 levels in the corn and barley groups were lower than those in other whole grain groups(P<0.05).Compared with the polished rice group and the fine noodle group,the concentration of TNF-α of the wheat,corn,millet and oat group was decreased,but the difference was not statistically significant(P>0.05),while the level of TNF-α in the barley group was lower than that in the polished rice group and the fine surface group(P<0.05).(2)Compared with the polished rice control group and the fine control group,the five whole grains could down-regulate the TLR4,TLR5,TNFα,and NFκB mRNA levels and the expression of IκBα,MyD88,TLR4 and NFκB proteins in the intestine(P<0.05).The comparison of the five whole grains showed that the regulation of TLR4 mRNA was lower in the corn group than in the wheat group and the oat group(P<0.05).The regulation effect of TNFα mRNA in the barley group and the millet group was lower than wheat,corn and oat group(P<0.05),The expression of TLR4 protein in maize group,barley group,millet group,and oat group was lower than that in the wheat group(P<0.05).(3)The effects of different types of whole grains on intestinal flora are different,and the abundance and composition of the main flora of the intestinal flora are different.In this experiment,bacteroidetes,proteobacteria,tenericutes,fusobacteria,and acidobacteria are the main bacteria that produce differences in different types of grains.Oats have the most obvious effect on improving intestinal flora,and oats can increase the expression of Bacillus and Bacteroides compared with the insulin resistance model group.(2)Study on different dose of oats on glucolipid metabolism and its mechanism.Through the first phase of animal testing,(1)Animal experimental results of different dosage oats show that the level of TG,FPG,HbAlc,HOMA-IR of four doses compared with refined rice control group,fine noodles control group(P<0.05),and the level of HDL-C increase obviously(P<0.05).The effect of the 25% oatmeal group is close to that of the polished rice noodle group.The effect of 50% oat group was similar to that of 100% oat group(P>0.05).Compared with insulin resistance model group,the four doses of oatmeal could reduce the LPS concentration(P<0.05),but there was no significant difference between the 25% oatmeal group and the polished rice noodle control group(P>0.05).The concentration of hs-CRP and IL-6 in the ointment group was lower than that in the polished rice noodle group(P<0.05).There was no significant difference between the four oatmeal groups(P>0.05).Compared with the polished rice noodle group,the TNF-α level in the 25% oat group was decreased,but the difference was not statistically significant(P>0.05),and the remaining dose oat group could reduce the TNF-α level(P<0.05).However,there was no significant difference between the different doses(P>0.05).(2)Compared with the high-fat control group and the polished rice control group,25%,50%,75%,and 100% oats could down-regulate TLR4,SREBP,ChREBP,and TNF-α mRNA levels and protein expression of p-IkBa,NF-κB,MyD88,TLR4 and SREBP1 c in liver tissue(P<0.05).(3)Different doses of oats have different effects on the intestinal flora,and the abundance and composition of the main flora of the intestinal flora are different.Oats can reduce the expression abundance of Clostridiales,increase the expression of bacteroides,and there is a dose-response relationship in intestinal flora.Conclusion:(1)Different types of whole grains all have insulin resistance effects in rats induced by a high-fat diet.The possible mechanism of improving insulin resistance by lowering blood lipids,regulating TLR4,TLR5 and NF-κB expression in the intestine,and improving the state of inflammatory factors.Long-term high-fat diets have an effect on the intestinal flora of rats.Feeding different types of whole grains can cause different changes in the structure and composition of the intestinal flora,and oats have the most significant effect on improving intestinal flora.The possible mechanism for improving the metabolism of glycolipids in whole grains is to improve the intestinal flora,reduce the penetration of LPS,improve the expression of key genes and proteins of glycolipid metabolism and inflammatory factors,thereby reducing the inflammatory level induced by the high-fat and high-cholesterol diet,improving the level of insulin resistance,and finally improving the glycolipid metabolism.The best effect on improving insulin resistance in whole grain is oats.(2)Different doses of oat can improve the insulin state of IR rats and have a doseresponse relationship.The mechanism of oat improving glycolipid metabolism could be the reducing of blood sugar and blood lipid levels in rats,reducing the expression of NF-κB in the liver,alleviate lipotoxicity and increasing insulin sensitivity,then improve insulin resistance and inflammatory state.Compared to other grains,oats affect the gut microbiota in a unique way,and feeding different doses of oat can also cause different changes in the structure of the gut flora.The 25% dose group has the effect of improving insulin status,but the improvement is limited.However,50% oats can significantly improve glycolipid metabolism and inflammatory factor levels after replacing polished rice noodles.With the increase of dose,there is a dose-effect relationship on blood sugar,blood lipids,inflammatory factors,etc.However,the different effect in this index is not obvious,the reason may be because oats need a long time to show their biological effects.