| AimThe incidence rate of acute and chronic complications of diabetes is increasing with the increasing incidence of diabetes at home and abroad.Diabetics are two to four times more likely to have a heart attack or stroke than nondiabetics.Dipeptidyl peptidase 4(DPP-4)inhibitors,as a new type of hypoglycemic drugs,have two-way regulation of ? function of islets of Langerhans,with definite hypoglycemic effect and low incidence of hypoglycemia.Hyperglycemia and high free fatty acids in diabetic patients make ?-cell function worsen gradually,aggravate the disorder of blood glucose and free fatty acids,lead to vicious circle,accelerate the process of atherosclerosis,and induce cardiovascular and cerebrovascular events.DPP-4 inhibitor has definite hypoglycemic effect and can protect ? cell function.At present,studies have shown that DPP-4 inhibitors may have cardiovascular and cerebrovascular protective effects.It is speculated that the possible mechanism is to improve blood lipid,block the activation/chemotaxis of monocytes,prevent the migration of monocytes and actin polymerization,so as to reduce the contents of visceral adipose tissue,megaphagocytes and inflammation,so as to play an anti atherosclerotic role.As one of the DPP-4 inhibitors,wiggletin can increase the insulin secreted by ?cells and effectively reduce blood sugar.As a hypoglycemic drug,wigletin can reduce the fat secretion in intestine,mobilize and burn fat during meal,and significantly reduce the levels of total cholesterol(TC),triglyceride(TG)and low-density lipoprotein(LDL)by reducing fasting lipolysis and apolipoprotein B-48.In addition,when used in patients with dyslipidemia due to nonalcoholic fatty liver disease,wiggletin can significantly improve body weight,body mass index(BMI),TC,TG and low-density lipoprotein cholesterol(LDL-C)levels.Because non-high density lipoprotein cholesterol,especially low density lipoprotein cholesterol,and triglycerides are risk factors of cardiovascular and cerebrovascular events,we think it is necessary to explore the role of viggliptin in liver lipid metabolism.At present,most researchers don't know whether the regulation of visgliptin on lipid metabolism is related to GLP-1,which needs to be further studied in vivo and in vitro.In this study,we used mouse model and HepG2 cell model to determine whether viggliptin can improve liver lipid,and tried to explore the possible mechanism of viggliptin treatment on lipid metabolism.Method1.Animal experiment:C57BL/6J mice of 6-8 weeks old male were purchased from Beijing weitonglihua company.They were raised in a clean environment and controlled by light(12 hours of light and dark cross).The indoor temperature was maintained at 22-23?.The feed was purchased from Beijing keaoxihe Feed Co.,Ltd.the common feed was 100%basic feed,14.09 kJ/g;the hot card of high-fat feed was 17.05 kJ/g,the formula was 82.7%basic feed+2%cholesterol+15%lard+0.3%sodium cholate.Viagliptin is from Novartis.It is dissolved in double distilled water and evenly mixed.It is administrated by gavage according to 50mg/kg.D.Mice not treated with wiggleptin were treated with double distilled water 10ml l kg.D by gavage.According to the experimental design,the mice were fed with general diet,high fat diet and high fat diet combined with wiggletin for 20 weeks.After anesthesia,blood was collected from the eyeball and centrifuged for preservation;part of the liver was frozen in liquid nitrogen for subsequent use,and part of the liver was fixed with 4%paraformaldehyde for morphological.2.Cell experimentHepG2 cells,the same biological activity as hepatocytes,were purchased from the cell bank of typical culture preservation Committee in Shanghai.In cell culture,MEM medium was used,10%fetal bovine serum(FBS),1%glutamine(gibico),100 U/ml penicillin and 0.1mg/ml streptomycin were added before use to prepare complete cell culture medium.The cell treatment reagent is provided by vegletin Novartis Co.,Ltd.;GLP-1 is purchased from Meilun biology.Western blotting used Rabbit anti HMGCR,CYP7A1,LDLR,SR-B1,ACAT2 to buy from Abcam;rat anti GAPDH to buy from Wuhan Sanying;mouse anti-2 and rabbit anti-2 to buy from Jackson.Free cholesterol(#E1016)and total cholesterol(#E1015)test kits were purchased from pulley company.Total free cholesterol Philippines(filpin)fluorescent staining kit was purchased from genemed Sciences inc.USA(GMS80059.1 v.A).Result1.Weight gain of mice in the treatment groupAfter one week of adaptive feeding,mice were divided into groups.After 12 weeks of general diet,high-fat diet and high-fat diet combined with vildagliptin,the weight of mice increased significantly(20.75±0.50g vs 24.08±0.49g,20.70±1.43 vs 29.81 ± 3.08g,21.38±1.20 vs,respectively)24.68±1.96g,P<0.01).The body weight of mice fed with high-fat diet was significantly higher than that of mice fed with ordinary diet(3.4±0.71g vs 9.1±2.25g,P=0.000).The body weight growth of mice treated with vildagliptin was significantly lower than that of mice not treated with vildagliptin(9.1 ± 2.25g vs 3.3±1.51g,P=0.000).2.Vildagliptin does not increase the risk of liver injuryCompared with the mice in the high fat diet group,the liver/body weight ratio decreased significantly(2.81 ± 0.62%vs 3.37±0.07%,P<0.05).The results showed that the serum ALT and AST concentrations of mice fed with high-fat diet were 37.22±4.51 U/L and 168.12 ± 21.76 U/L,respectively,and the concentrations of mice fed with high-fat diet were 43.24 ± 16.79 U/L and 180.19 ±36.68 U/L,respectively(P>0.05).3.Vildagliptin improves glucose metabolism in mice with high fat dietThere was no significant difference in blood glucose(7.46 ± 1.22mmol/L vs 7.70± 1.34mmol/L)between the two groups(P>0.05).The results of oral glucose tolerance test showed that the blood glucose of mice in the high fat diet group decreased at 30,60,90 and 120 minutes(9.75±1.10 vs.11.03± 1.73,9.01±0.97 vs.10.157±1.45,8.48±1.10 vs.9.99±1.76,8.09±1.11 vs.9.49±1.33,P 0.04,0.03,0.02 and 0.02,respectively).The results of intravenous insulin test showed that there was a decrease(172.91 ±43.28 nIU/ml vs.209.93±38.52 nIU/ml)in the high fat diet group,but there was no statistical significance(P=0.11).HOMA-IR in the high fat diet group was slightly higher than that in the general diet group(0.07±0.03 and 0.06±0.02,P=0.04,respectively).After the application of wigglett,it was decreased(0.06±0.02,P=0.04).4.Vildagliptin reduces the levels of LDL in mice with high fat dietThe blood samples of the mice treated with wigglettin showed that the LDL(0.47±0.07mmol/L)of the mice treated with wigglettin was significantly lower than that of the mice fed with high fat diet(the latter concentration was 0.70±0.17mmol/L,P<0.01),and the TC was decreased(3.99±0.64mmol/L vs 3.87±0.97mmol/L,P>0.05).The serum HDL and TG levels were not significantly different from those of the mice fed with high fat diet(2.73±0.47mmol/L vs,respectively)2.36 ±0.46mmol/1,0.25±0.09mmol/l vs 0.23±0.05mmol/l),P equals to 0.21 and 0.21,respectively.5.Vildagliptin reduces liver lipid content in mice fed with high fat dietHE staining showed that the liver structure of different intervention groups were normal.The content of liver lipid in different groups of mice showed that FC in the modified group was significantly lower than that in the high fat diet group(56.52±12.70 umol/g protein,P=0.031),but TC and TG were only decreased after the adjustment(116.7 ± 44.38 umol/g protein,179.35 ± 198.41 umol/g protein respectively),had no statistical significance(P=0.006,0.51)to significantly reduce the total cholesterol in the liver(P<0.05).Western blotting showed that the expression of HMGCR and CYP7A1 in the high fat diet with vildagliptin group was higher than that in the high fat diet group.6.Vildagliptin promotes the expression of CYP7A1 and HMGCRThere was no significant difference in free cholesterol,total cholesterol and triglyceride content between the two groups(P>0.05).Western blotting showed that the protein expression of CYP7A1 and HMGCR was increased by vildagliptin,suggesting that vildagliptin could promote the expression of CYP7A1 and HMGCR.Conclusion1.Vildagliptin reduced the abnormal glucose tolerance and insulin resistance of mice induced by high fat diet;2.Vildagliptin partially improved the abnormal blood lipid and liver lipid caused by high fat diet in mice;3.Vildagliptin promotes the expression of HMGCR and CYP7A1 simultaneously,which is independent of GLP-14.Vildagliptin did not change the lipid of HepG2 cells.AimDiabetes is a chronic non infectious disease,which brings great economic burden to society and family.Both in China and in the world,the number of patients with diabetes is increasing.As we all know,the abnormal metabolism of insulin will lead to the abnormal function of thyroid.Similarly,the imbalance of thyroid hormone will also affect the metabolism of insulin.Studies have shown that diabetic patients with hyperthyroidism are more likely to suffer from hyperglycemic crisis and recurrent hypoglycemia.The serum levels of free triiodothyronine(FT3)and thyroid stimulating hormone(TSH)were higher in patients with type 2 diabetes mellitus(T2DM),and the prevalence of primary hypothyroidism was higher.In glucose metabolism,patients with hyperthyroidism have higher glucose levels under fasting conditions.China is one of the countries with the largest population.The prevalence of diabetes mellitus and prediabetes in adults are 11.6%and 50.1%respectively.There have been studies on the prevalence of thyroid diseases in diabetic patients.However,the subjects of study are limited to diabetic patients.The prevalence of thyroid diseases in prediabetes is less concerned.The risk factors of thyroid diseases in patients with abnormal glucose metabolism are still unknown Unknown.In order to determine the prevalence of thyroid disease in different blood glucose status,this study investigated the prevalence of thyroid disease in Ningyang area of Shandong Province,analyzed the prevalence of cardiovascular cerebrovascular disease(CCVD)in subclinical hypothyroidism population with abnormal blood glucose status,and preliminarily explored the risk factors of thyroid disease in different glycemic glucose metabolism status.MethodObject:This study is a large-scale,observational,cross-sectional study based on the population.The research object is from the action research conducted in China in 2011,and the research scheme has been approved by the ethics committee of Shanghai Jiaotong University.The subjects who met the inclusion criteria were collected through one-stop epidemiological survey,including filling in questionnaires,receiving physical examination and collecting overnight fasting blood samples.The subjects who denied the history of diabetes were also required to collect oral glucose tolerance for 2 hours.The criteria for inclusion and exclusion were as follows:(1)Inclusion criteria:Residents who have lived in Ningyang County of Shandong Province for more than 5 years and over the age of 18 agree to participate in the screening and sign the informed consent.(2)Exclusion criteria? Within six months of pregnancy or postpartum;? Take drugs(including thyroid hormone and antithyroid drugs)that may affect thyroid function in the past 3 months;? Suffer from diseases that may affect thyroid function,such as mental diseases,acute cardiovascular and cerebrovascular diseases,malignant tumors,and hypothalamic pituitary diseases.?Severe hepatic and renal insufficiency(see diagnosis of disease);? Lack of important data related to the study,such as serum glucose or thyroid function results;?Cardiovascular diseases,including stroke,myocardial infarction(MI),hypertension,and coronary atherosclerotic heart disease(CHD),were identified in the population with normal blood glucose;Methods:The investigators,with medical background,were trained in standardization,and conducted questionnaire,physical examination,blood sample collection and testing after passing the examination.Procedure:In order to reduce the information bias caused by different judgment standards or investigation methods in the process of data collection,the investigators have medical background,all of them have received standardized training,and after passing the examination,they have carried out questionnaire,physical examination,blood sample collection and testing.All serum samples were cryopreserved and transported under-20? in the refrigerator,and the related tests were completed in the clinical laboratory of Shandong provincial hospital.Electrochemiluminescence methods(Cobas E601;Roche,Basel,Switzerland)were used to detect thyroid function indexes,including thyrotropin,free thyroxine,free triiodothyronine,Beckman chemistry analyzer au5800 system,Beckman Coulter,Tokyo,Japan)to detect triglycerides,low-density lipoprotein,high-density lipoprotein,total cholesterol,fasting glycemic glucose,postprandial blood glucose,alanine aminotransferase,aspartate aminotransferase,creatinine and glycosylated hemoglobin.The intraassay and inter assay differences of the above serological tests were all controlled below 5%.According to the 2010 standard of American Diabetes Association,Chinese thyroid disease diagnosis and treatment guide and the reference range of clinical normal laboratory,the disease diagnosis was carried out.Statistical method:SPSS version 22.0(for windows,Chicago,IL,USA)was used for statistical analysis.The classified variable is described by frequency(percentage),and the numerical variable of normal distribution is represented by mean ± SD.Chi square test(classified variable),t test or analysis of variance(numerical variable)are selected according to the situation.Multiple logistic regression analysis was used to determine which glycemic glucose level was most related to thyroid disease.The cut point of the significant difference was p<0.05.Result:1.Basic clinical characteristics of subjectsIn this study,10860 subjects were enrolled,including 4437 males(40.86%)and 6423 females(59.140%),with an average age of 55.37±8.82 years.Among these subjects,13.63%of them had normal blood glucose,58.80%of them had pre diabetes and 27.57%of them had diabetes.2.The prevalence of thyroid diseases was higher in subjects with abnormal blood glucoseThe prevalence of thyroid disease was 13.43%(n=1459).The prevalence of thyroid disease in the population with abnormal blood glucose was higher than that in the population with normal blood glucose(14.28%vs.8.11%,P<0.001).Among the patients with thyroid diseases,subclinical hypothyroidism is the most,with 1267 cases(86.84%),followed by hypothyroidism(7.47%),subclinical hyperthyroidism(3.63%)and hyperthyroidism(2.06%).Furthermore,subclinical hyperthyroidism and subclinical hypothyroidism classified into hypothyroidism and hyperthyroidism statistics,the proportion of different thyroid diseases in people with abnormal blood glucose was significantly different from that in people with normal blood glucose(P<0.001).3.Gender,age and HDL are the risk factors of subclinical hypothyroidism indiabetic subjectsAmong the subjects with diabetes mellitus,the proportion of women with subclinical hypothyroidism was higher than that with normal hypothyroidism(61.36%vs.54.26%,P<0.05);the former was younger than the latter(56.68±8.21 years vs.57.84±8.63 years,P<0.05).The TG level of diabetic patients with subclinical hypothyroidism(2.43±3.38 mmol/L)was higher than that of diabetic patients with normal hypothyroidism(1.80±1.40 mmol/L,P<0.01).There was also a significant difference between the two groups in TC(5.61±1.38 mmol/L vs.5.30±1.23 mmol/L)and LDL(3.34 ± 1.02 mmol/L vs.3.16±0.95 mmol/L)(P<0.01),but there was no difference in HDL between the two groups(1.36±0.36 mmol/L vs.1.38±0.35 mmol/L,p=0.21).There was no difference in HbAlc,FBG and PPG between diabetic patients with subclinical hypothyroidism and those with normal hypothyroidism.Women and age were risk factors for diabetes,OR was 2.53[95%CI(1.99-3.27)]and 1.01[95%CI(1.00-1.03)],respectively.Diabetic patients with low HDL were more likely to develop subclinical hypothyroidism[OR 0.562,95%CI(0.388-0.815)].4.Sex and age are the risk factors of subclinical hypothyroidism in prediabetesAmong the pre diabetes subjects,the proportion of women with subclinical hypothyroidism was higher than that with normal hypothyroidism(63.91%vs.58.28%,P<0.01).The former was older than the latter(56.82±8.18 years vs.54.84± 8.71 years,P<0.01).HbA1c,TG,TC and LDL of pre diabetic patients with subclinical hypothyroidism were higher than those with normal hypothyroidism(5.87±0.29%vs.5.82±0.31%,respectively),1.50±1.03 mmol/L vs.1.37 ± 1.01 mmol/L,5.28±1.18 mmol/L vs.5.07 ± 1.11 mmol/L,3.17 ± 0.93 mmol/L vs.3.00±0.87 mmol/L,P<0.01),HDL was lower than the latter(1.42± 0.33 mmol/L vs.1.46±0.37 mmol/L,P<0.01).Women were the risk factors of prediabetes,OR was 2.55[95%CI(2.15-3.02)].With the increase of age,the risk of subclinical hypothyroidism in prediabetes increased significantly,OR was 1.01,95%CI(1.00-1.02).5.The highest prevalence of CCVD in diabetes mellitus with subclinical hypothyroidismThe incidence of CCVD in diabetic patients with subclinical hypothyroidism(33.97%)was significantly higher than that in non diabetic patients(26.51%),the difference was statistically significant(P<0.01),and also higher than that in pre diabetic patients(33.97%vs.16.78%;P<0.01).The incidence of CCVD in pre diabetic patients with subclinical hypothyroidism(16.78%)was higher than that in non diabetic patients(15.87%),but there was no statistical difference(P=0.56).Conclusion:1.The prevalence of thyroid disease in the patients with glucose metabolism disorder was higher than that in the normal people.2.Among the patients with the disorder of glucose metabolism,subclinical hypothyroidism was the most common(86.84%),followed by hypothyroidism(7.47%),subclinical hyperthyroidism(3.63%),and hyperthyroidism was the least common(2.06%).3.Women and age are the risk factors of subclinical hypothyroidism.In diabetic patients,the decrease of HDL is the risk factor of subclinical hypothyroidism.4.The prevalence of CCVD in diabetic patients with subclinical hypothyroidism is higher than that in patients with simple diabetes. |
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