| In the research,there are two parts to study the relationship between T2 DM with MCI and sex hormone signaling pathway.First part Theoritic exploration Explain the etiology,pathogenesis,syndrome differentiation and treatment of diabetic cognitive disorder by the perspective of traditional Chinese medicine,the tutor's views and my own research.Second part Scientific research Objective: To observe the learning and studying ability,sex hormone,and the activity of Ch AT,Apo E and A?1-42 in hippocampal of type 2 diabetes mice.Thus clear the effect and mechanism of refreshing and puzzling decoction together with rosiglitazone on developing hippocampus memory impairment of type 2 diabetes mice and provide new scientific basis for explaining “Pancreas(spleen)-brain-kidney” pathogenesis theory of MCI with T2 DM.Methods: 10 weeks old grade male KM mice were given a basic diet as the normal control group;10-week-old SPF level male KKay mice were given a high-sugar and high-fat diet.All of the mice were provided by the Chinese Academy of Medical Sciences Institute of Experimental.6 weeks later,when the random tails assay blood glucose of KKay was more than 13.9mmol/L,the animal model was set up successfully.Then randomly divided them into model control group,positive control group,TCM group,and TCM OVX(ovariectomized)group.Fed Positive group with Laci Staw(150mg·kg-1)and rosiglitazone maleate tablets(0.003 g · kg-1)oral administration daily,while TCM group and TCM OVX group were fed with rosiglitazone maleate piece(0.003 g · kg-1)and Xingnaoyizhi agent(7.2g · kg-1).Fed the normal control group and model control group with an equal volume of saline daily.The gavage lasted for 8 weeks.We used Morris water maze to test the study and memory capacity of the mice both before and after giving medicine;enzyme linked immunosorbent assay(ELISA)to observe the protein expression of sex hormone levels in mice's plasma;QPCR to observe Ch AT and Apo E m RNA levels in mice's hippocampus;Western Blotting to observe A?1-42 level in mice's hippocampus.Results: 1,General status and condition of among the groups: The mice in the normal control group were all in good condition with white hair color with a slight sheen,normal drinking water,and daily action sensitive active;With the increase of age,different degrees of burnout,dry hair color,unresponsive,listlessness,polyphagia polyuria,and slow appear in the mice of positive group,TCM group and TCM OVX group,especially the model control group;2,Experimental results of animal behavior by Morris water maze(1)Results on the fifth day before administration showed that,compared with the normal control group,there were significantly higher latency and lower platform quadrant probability in model control group,positive control group,TCM group,and TCM OVX group(P<0.05);compared with TCM OVX group,the other KKAy groups had more lower latency with statistically significant difference(P<0.05);(2)Compared with the results on the fifth day before administration,positive control group and TCM group had more higher latency and lower platform quadrant probability with statistically significant difference(P<0.05);TCM OVX group had slightly higher latency and lower platform quadrant probability with no statistically significant difference(P>0.05);(3)Results on the fifth day after administration showed that,compared with the normal control group,there were significantly higher latency in model control group,positive control group,TCM group,and TCM OVX group,and lower platform quadrant probability in model control group,positive control group,and TCM OVX group with statistically significant difference(P<0.05);compared with model control group,there were significantly lower latency and higher platform quadrant probability in normal control group,positive group,and TCM group with statistically significant difference(P< 0.05);compared with positive group,there were slightly higher platform quadrant probability in TCM group with no statistically significant difference(P > 0.05);compared with TCM group,there were significantly higher latency and lower platform quadrant probability in TCM OVX group with statistically significant difference(P <0.05);3,Effects on blood glucose and body weight(1)Blood glucose Compared with the normal control group before and after administration,groups of KKAy mice's blood glucose was significantly increased with statistically significant difference(P<0.05);compared with before administration,mice of positive control group,TCM group,and TCM OVX group had much more lower blood glucosewith statistically significant difference(P < 0.05);after administration,compared with the model control group,mice of positive control group,TCM group,and TCM OVX group had much more lower blood glucosewith statistically significant difference(P<0.05);there is no statistically significant difference between other groups(P>0.05);(2)Body weight Compared with the normal control group before and after administration,groups of KKAy mice's body weight was significantly increased with statistically significant difference(P < 0.05);compared with before administration,all groups had slightly higher body weight,but with no statistically significant difference(P>0.05);4,Changes of plasma estrogen levels in mice(1)Compared with the normal control group,model control group and TCM OVX group had much lower level of Estradiol(E2)in plasma with statistically significant difference(P<0.05);compared with the model control group,TCM group had much higher level of E2 in plasma with statistically significant difference(P < 0.05);compared with the positive group,TCM group had much higher level of E2 in plasma with statistically significant difference(P < 0.05);compared with TCM group,TCM OVX group had much lower level of E2 in plasma with statistically significant difference(P<0.05);(2)Compared with the normal control group,model control group and TCM OVX group had much lower level of progesterone(P)in plasma with statistically significant difference(P < 0.05);compared with the model control group,TCM group had much higher level of P in plasma with statistically significant difference(P<0.05);compared with the positive group,TCM OVX group had much lower level of P in plasma with statistically significant difference(P < 0.05);compared with TCM group,TCM OVX group had much lower level of E2 in plasma with statistically significant difference(P<0.05);(3)There was no significant difference in serum testosterone(T)between the groups(P>0.05);(4)Compared with the normal control group,model control group,positive group,and TCM OVX group had much higher level of follicle stimulating hormone(FSH)in plasma with statistically significant difference(P < 0.05);compared with the model control group,TCM group had much lower level of FSH in plasma with statistically significant difference(P < 0.05);compared with the positive group,TCM group and TCM OVX group had lower level of FSH in plasma,bu with no statistically significant difference(P> 0.05);compared with TCM group,TCM OVX group had higher level of FSH in plasma,bu with no statistically significant difference(P>0.05);(5)Compared with the normal control group,model control group,positive group,and TCM OVX group had much higher level of luteinizing hormone(LH)in plasma with statistically significant difference(P < 0.05);compared with the model control group,TCM group had much lower level of LH in plasma with statistically significant difference(P<0.05);compared with the positive group,TCM group had lower level of FSH in plasma with statistically significant difference(P<0.05);compared with TCM group,TCM OVX group had higher level of FSH in plasma with statistically significant difference(P<0.05);(6)Compared with the normal control group,model control group,positive group,TCM group and TCM OVX group had much higher level of prolactin(PRL)in plasma with statistically significant difference(P < 0.05);compared with the model control group,positive group,TCM group and TCM OVX group had much higher level of prolactin(PRL)in plasma,bu with no statistically significant difference(P>0.05);5,Compared with the normal control group,model control group,and TCM OVX group had much lower Ch AT m RNA expression level in brain hippocampus with statistically significant difference(P<0.05);compared with the model control group,positive group,and TCM group had much higher Ch AT m RNA expression level in brain hippocampus with statistically significant difference(P < 0.05);compared with TCM group,TCM OVX group had much lower Ch AT m RNA expression level in brain hippocampus with statistically significant difference(P<0.05);6,Compared with the normal control group,groups of KKAy mice's Apo E m RNA expression level in brain hippocampus was significantly decreased with statistically significant difference(P < 0.05);while there was no statistically significant difference between groups of KKAy mice(P>0.05);7,Compared with the normal control group,groups of KKAy mice's A?1-42 expression level in brain hippocampus was significantly decreased with statistically significant difference(P<0.05);compared with the model control group,positive group and TCM OVX group had much lower A ? 1-42 expression level in brain hippocampus with statistically significant difference(P<0.05);compared with positive group,TCM group had much lower A ? 1-42 expression level in brain hippocampus with statistically significant difference(P < 0.05);compared with TCM group,TCM OVX group had much higher A?1-42 expression level in brain hippocampus with statistically significant difference(P<0.05).Conclusion: 1,Effect of Xingnao Yizhi mixture on sex hormones: the cognitive function of 18 week old T2 DM mice has been shown to be impaired and has a significant sex hormone disorder with decreasing of E2 and P,and increasing of FSH,LH and PRL.The most obvious change was E2,and the change of T was not obvious.Xingnao Yizhi mixture can better correct the level of sex hormone disorder to improve E2 and P,and reduce LH and FSH.And this effect was superior to that of castrated mice in T2 DM mice.2,Effect of Xingnao Yizhi mixture on Ch AT,Apo E m RNA and A?1-42 expression level in hippocampus(1)Type 2 diabetes mellitus with cognitive dysfunction KKAy mice had high level expression of A?1-42 protein.Xingnao Yizhi mixture and Laci Staw can effectively reduce its expression level of A?1-42 protein,especially better for adults mice.The expression level of A?1-42 in the hippocampus of mice is directly proportional to the level of serum luteinizing hormone and inversely proportional to the level of serum estradiol and progesterone;(2)Type 2 diabetic cognitive dysfunction may be caused by the excessive lowering m RNA expression level of Ch AT in brain hippocampus.Xingnao Yizhi mixture and Laci Staw can improve the expression of Ch AT m RNA level in hippocampus of brain.The expression level of Ch AT m RNA may be directly proportional to serum estradiol and inversely proportional to serum luteinizing hormone level.Effect of Xingnao Yizhi mixture on the rapid aging of type 2 diabetic cognitive dysfunction in mice Ch AT m RNA expression was inhibited by sex hormone disorder,so as the recovery of learning and memory function.(3)Study on effect of Xingnao Yizhi mixture on cognitive dysfunction in mice with type 2diabetes cognitive function1)The cognitive function of KKAy mice decreased significantly at 18 weeks of age.The cognitive function of rapidly aging type 2 diabetic mice was significantly lower than that of the adult mice,and the difference may be the formation of abnormal levels of sex hormones;2)After treating with Lacita and Xingnao Yizhi mixture,the cognitive function of type 2diabetic mice was significantly improved,but it could not reach the level of normal mice;3)After treating with Xingnao Yizhi mixture,the learning and memory ability of mice with rapidly aging type 2 diabetes was improved,but was lower than that in normal mice and adult mice with type 2 diabetes.This difference may be due to the drug can not effectively regulate the disorder of sexual hormone secretion caused by;4)Xingnao Yizhi mixture might have improved cognitive dysfunction in type 2 diabetic mice through increasing levels of estradiol and progesterone in plasma and Ch AT?Apo E m RNA expression in hippocampus,and reducing the levels of A?1-42 in hippocampus and plasma FSH. |
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