摘要
目的:研究给予2型糖尿病大鼠瑞格列奈+二甲双胍、罗格列酮+二甲双胍、胰岛素+二甲双胍干预后,各组大鼠空腹血清visfatin水平与体重、血糖、胰岛素、胰岛素抵抗、脂代谢紊乱及大鼠胰岛β细胞凋亡之间的关系。
方法:(1)采用高热量饲料8周+单次腹腔注射小剂量链尿佐菌素(STZ)制备2型糖尿病大鼠模型,以1周后空腹血糖≥11.1mmol/L为成模标准(n=28),将糖尿病大鼠分别予以瑞格列奈+二甲双胍(B,n=7)、罗格列酮+二甲双胍(C,n=7)、胰岛素+二甲双胍(D,n=7)、生理盐水(E,n=7)干预8周;同时采用高热量饲料8周制备单纯肥胖组大鼠(A,n=8)模型;并设定正常大鼠为对照组(NC,n=10)。(2)2型糖尿病各治疗组大鼠、未治疗组大鼠、单纯肥胖组及对照组大鼠为研究对象,测量大鼠体重;酶联免疫法(EIA)测定空腹血清visfatin浓度,并测定空腹血糖、甘油三酯、总胆固醇、低密度脂蛋白胆固醇、高密度脂蛋白胆固醇、胰岛素,并计算胰岛素抵抗指数。(3)TUNEL法检测大鼠胰岛β细胞凋亡率。
结果:(1)糖尿病各治疗组和未治疗组FPG、FINS、HOMA-IR、TC、TG、LDL、FFA、visfatin及体重较NC组均显著升高(P＜0.01),FPG较单纯肥胖组显著升高(P＜0.01),HDL较NC组均明显下降(P＜0.01);给予不同治疗后(B-D组)FPG、FINS、HOMA-IR较未治疗组(E组)明显下降(P＜0.05),罗格列酮组+二甲双胍(C)组FFA较其他各组明显下降(P＜0.05),而visfatin较其他各组明显升高(P＜0.05),瑞格列奈+二甲双胍组、胰岛素+二甲双胍组、未治疗组和单纯肥胖组间visfatin无明显差异(P＞0.05)。(2)Pearson相关分析和多元回归分析显示:单纯肥胖组(A)和瑞格列奈+二甲双胍组(B)的TG、LDL、FFA及体重与visfatin呈正相关(P＜0.05),FFA对单纯肥胖组空腹血清visfatin浓度影响最显著(r=0.964,P=0.0),被引入回归模型中。罗格列酮+二甲双胍组(C)TG和体重与visfatin呈正相关(P＜0.05);胰岛素+二甲双胍组(D)TG、LDL、FFA、体重与visfatin呈正相关(P＜0.05),HDL与visfatin呈负相关(P＜0.01);未治疗组(E)FPG、TG、LDL、FFA、体重与visfatin呈正相关(P＜0.05),HDL与visfatin呈负相关(P＜0.01),B-E组体重对空腹血清visfatin浓度影响最显著(r=0.964,P=0.0;r=0.927,P p=0.003;r=0.967,P=0.004;r=0.917,P=0.014)被引入回归模型。(3)各组大鼠胰岛β细胞凋亡率的比较:与正常对照组的胰岛β细胞凋亡率(16±4)%相比,单纯肥胖组(25±7)%、瑞格列奈+二甲双胍(29±6)%、罗格列酮+二甲双胍(22±4)%、胰岛素+二甲双胍(28±5)%、2型糖尿病未治疗组(32±9)%的胰岛β细胞凋亡率均明显增加(P＜0.01);单纯肥胖组、瑞格列奈+二甲双胍、胰岛素+二甲双胍、罗格列酮+二甲双胍的胰岛β细胞凋亡率较2型糖尿病未治疗组均明显减少(P＜0.05);罗格列酮+二甲双胍组的胰岛β细胞凋亡率较其他各治疗组明显减少(P＜0.05);罗格列酮+二甲双胍组的胰岛β细胞凋亡率与血清visfatin浓度呈明显负相关(P＜0.05)。
结论:(1) 2型糖尿病和单纯肥胖组大鼠血浆visfatin水平均升高;给予不同干预后除罗格列酮+二甲双胍组血浆visfatin水平增高外,其余各治疗组血浆visfatin水平无明显变化;且罗格列酮+二甲双胍组胰岛β细胞凋亡率与血浆visfatin水平成负相关,提示PPARγ核受体可能参与调控visfatin的表达和分泌活动,visfatin可能是PPARγ的激动剂药物治疗2型糖尿病的作用靶点之一。
(2)血浆visfatin水平与脂代谢和体重密切相关,与胰岛素抵抗无明显关系。
(3)罗格列酮联合二甲双胍治疗能更好的改善胰岛素抵抗、保护胰岛β细胞,是目前较为理想的治疗初发肥胖型2型糖尿病的治疗选择。
AIM: To investigate the change of fasting plasma visfatin level to give different interventions(repaglinide+dimethylbiguanide, rosigLitazone +dimethylbiguanide, insulin +dimethylbiguanide)in rats with type 2 diabetes, and to study the relationship of plasma visfatin level to FPG, FINS, HOMA-IR, TC, TG, HDL, LDL, FFA and body weight;To detect The frequency ofβcell apoptosis and analysis the relationship of plasma visfatin level to it.
Methods: (1)Use high-sucrose and high-fat diet+ STZ(35mg/kg) inducing wistar rats,develop rats model with type 2 diabetes melitus by means of FPG≥11.1mmol/L(DM,n=28),and then to give repaglinide+dimethylbiguanide(B, n=7), rosigLitazone+dimethylbiguanide(C, n=7), insulin+dimethylbiguanide(D, n=7), NS(E, n=7) in DM about 8 weeks;Use only high-sucrose and high-fat diet develop obese rats model. (2) Use enzyme-linked immunoassay(EIA) to measure fasting plasma visfatin level. Fasting blood glueose(FBG), triglyeeride(TG), totaleholesterol(TC), low-density lipoprotein cholesterol(LDL-c), high-density lipoprotein cholesterol(HDL-c), fasting Serum insulin(FINS) were measured in all of the subjeets, Use HOMA-IR estimate insulin sensitivity.(3) The apoptoticβcells in islets were detected and quantified by the TUNEL technology(labeling of DNA strand breaks).
Results: (1) It is found that FINS, HOMA -IR,TC,TG, HDL, LDL, FFA, visfatin and body weight in diabetes rats and obese rats were significantly increased (P<0.01) and HDL were significantly reduced (P<0.01) compared with NC; FPG in diabetes rats were significantly increased (P<0.01) compared with NC and obese rats; FPG, FINS, HOMA-IR in B-E groups were significantly reduced (P<0.05) compared with F; FFA in C group were significantly reduced (P<0.05) and visfatin level were significantly increased (P<0.05) compared with other groups. (2) Correlation and multiple regression analysis shows that: The fasting serum visfatin of A and B group was positively correlated with viseus fat weight、TG、LDL and FFA (P<0.05);the FFA seemed to be the best variable in predicting obese rats visfatin level (r=0.964, P =0.0) ; The fasting serum visfatin of C group was positively correlated with viseus fat weight and TG (P< 0.05); The fasting serum visfatin of D group was positively correlated with viseus fat weight、TG、LDL and FFA (P<0.05), and negatively correlated with HDL (P<0.01); The fasting serum visfatin of E group was positively correlated with FPG、TG、LDL、FFA and fat weight (P<0.05), and negatively correlated with HDL (P<0.01); the fat weight seemed to be the best variable in predictingB-E group rats visfatin level (r=0.964, P =0.0; r =0.927, P =0.003; r =0.967, P =0.004; r =0.917, P =0.014) .(3) The frequency ofβcell apoptosis was higher in A-E group rats than in NC [(25±7), (29±6), (22±4), (28±5) , (32±9)vs(16±4), P <0.001]; The frequency of B cell apoptosis was higher in E group rats than in A-D group (P<0.001); There was no difference in frequency ofβcell apoptosis among A-D group (P >0.05); The fasting serum visfatin of rosigLitazone+dimethylbiguanide(C )group was negatively correlated with frequency ofβcell apoptosis (P <0.05) ; There was no difference in correlation of fasting serum visfatin and frequency ofβcell apoptosis in other groups (P >0.05) .
Conclusions: (1) It is found that fasting serum visfatin in diabetes rats and obese rats were significantly increased ; There was no difference of fasting serum visfatin level in intervention groups excluding rosigLitazone+dimethylbiguanide(C)group, and the fas -ting serum visfatin of rosigLitazone+dimethylbiguanide(C)group was negatively corr -elated with frequency ofβcell apoptosis, It is possible that PPARr nuclear receptor joins in the express and secretory activity of visfatin. visfatin, which is close correlated with glucolipid metabolism and body weight,may be one of the targets of PPARr's excitomoter drug treat DM. (2) Fasting serum visfatin level is close correlated with glucolipid metabolism and body weight,but isn't correlated with insulin resistance.(3) rosigLitazone+dimethylbiguanide can improve insuliu resistance and protect pancreatic beta cells more than others,so it is a better choice to treat the patient of type 2 diabetes mellitus.
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