摘要
目的探讨光甘草定(GLA)对2型糖尿病(T2DM)的缓解作用及机制。方法将36只清洁级Sprague-Dawley大鼠随机分为对照(CON)组、GLA对照(CON-GLA)组、糖尿病(DM)组、低剂量GLA(GLA-L)组、高剂量GLA(GLA-H)组和二甲双胍(MET)组。CON组和CON-GLA组大鼠给予正常饲料喂养;DM组、GLA-L组、GLA-H组和MET组大鼠给予高糖高脂饲料喂养,连续喂食3周制备T2DM大鼠。然后,CON组大鼠给予正常饲料并腹腔注射0.5 mL二甲基亚砜(DMSO);CON-GLA组大鼠给予正常饲料并腹腔注射0.5 mL GLA(40 mg·kg^(-1));DM组大鼠给予高糖高脂饲料并腹腔注射0.5 mL DMSO;GLA-L组大鼠给予高糖高脂饲料并腹腔注射0.5 mL GLA(20 mg·kg^(-1));GLA-H组大鼠给予高糖高脂饲料并腹腔注射0.5 mL GLA(40 mg·kg^(-1));MET组大鼠给予高糖高脂饲料并腹腔注射MET(150 mg·kg^(-1));每组大鼠均每2日给药1次,持续28 d。分别于给药第0、7、14、21、28天禁食10 h后测量大鼠体质量、空腹血糖(FBG),最后一次给药后1 d行葡萄糖耐量试验(OGTT),并计算时间-血糖曲线下面积(AUC);应用酶联免疫吸附法检测空腹胰岛素(FINS)和糖化血红蛋白(HbA1c)水平,计算胰岛素抵抗指数(HOMA-IR)和胰岛β细胞功能指数(HOMA-β);同时应用酶标仪检测血清中丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)水平,油红O染色观察大鼠肝脏脂肪沉积情况,并检测各组大鼠肝脏组织中三酰甘油(TG)和总胆固醇(TC)水平;采用Western blot法检测大鼠肝脏糖代谢不同途径中的关键限速酶丙酮酸羧化酶(PC)、葡萄糖-6-磷酸酶(G6Pase)、柠檬酸合酶(CS)和异柠檬酸脱氢酶2(IDH2)蛋白的表达。结果给药第0、7、14、21、28天时,DM组、GAL-L组、GAL-H组、MET组大鼠体质量均显著低于CON组(P<0.05);给药第14、21、28天,GAL-L组、GAL-H组、MET组大鼠体质量显著高于DM组,GAL-H组和MET组大鼠体质量显著高于GAL-L组(P<0.05)。给药第0、7、14、21和28天,DM组、GAL-L组大鼠FBG显著高于CON组与CON-GLA组(P<0.05);给药第0、7、14天,GLA-H组和MET组大鼠FBG显著高于CON组与CON-GLA组(P<0.05);给药第7、14、21和28天,GLA-L组、GLA-H和MET组大鼠FBG显著低于DM组(P<0.05)。给药第21、28天后,GLA-H组和MET组大鼠FBG显著低于GLA-L组(P<0.05)。与DM组相比,CON组、CON-GLA组、GLA-L组、GLA-H组和MET组大鼠血糖在30、60、120 min时均显著降低(P<0.05)。OGTT试验中GLA-L组大鼠血糖值随着时间不断降低,GLA-L组大鼠血糖值在30、60、120 min时显著高于CON组(P<0.05)。GLA-H组和MET组大鼠血糖随着时间变化的趋势与GLA-L组相似,GLA-H组与MET组大鼠血糖仅在30、60 min时血糖值显著高于CON组,120 min时GLA-H组和MET组大鼠血糖值显著低于GLA-L组(P<0.05);且GLA-H组和MET组大鼠血糖值与CON组比较差异无统计学意义(P>0.05)。GLA-H组与MET组大鼠血糖值在OGTT试验中各个时间点比较差异无统计学意义(P>0.05)。DM组、GLA-L组和GLA-H组大鼠的血糖AUC显著高于CON组和CON-GLA组(P<0.05);GLA-L组、GLA-H组和MET组大鼠血糖AUC均显著低于DM组(P<0.05);GLA-H组和MET组大鼠的血糖AUC显著低于GLA-L组(P<0.05);GLA-H组与MET组大鼠的血糖AUC比较差异无统计学意义(P>0.05)。与DM组相比,CON组、CON-GLA组、GLA-L组、GLA-H组和MET组大鼠HbA1c均显著降低(P<0.05)。GLA-H组和MET组大鼠HbA1c水平显著低于GLA-L组(P>0.05)。GLA-L组大鼠FINS显著高于CON组和CON-GLA组(P<0.05)。GLA-H组与CON组大鼠FINS和HOMA-IR比较差异无统计学意义(P>0.05)。与DM组相比,CON组、CON-GLA组、GLA-L组、GLA-H组和MET组大鼠FINS和HOMA-IR均显著降低(P<0.05)。CON组、CON-GLA组、GLA-L组、GLA-H组和MET组大鼠HOMA-β显著高于DM组(P<0.05)。CON组、CON-GLA组、GLA-L组、GLA-H组和MET组之间大鼠HOMA-β比较差异无统计学意义(P>0.05)。油红O染色结果显示,CON组和CON-GLA组大鼠肝脏脂类较少,DM组大鼠肝脏脂类大量聚积;GLA-L组和GLA-H组大鼠肝脏脂类聚积逐渐减少,但仍多于CON组和CON-GLA组。CON组与CON-GLA组大鼠肝脏TC和TG水平比较差异无统计学意义(P>0.05);DM组、GLA-L组、GLA-H组和MET组大鼠肝脏TC水平显著高于CON组和CON-GLA组(P<0.05)。GLA-H组和MET组大鼠肝脏TG水平与CON组和CON-GLA组比较差异无统计学意义(P>0.05);DM组和GLA-L组大鼠肝脏TG水平显著高于CON组和CON-GLA组(P<0.05)。GLA-L组、GLA-H组和MET组大鼠肝脏TC和TG水平显著低于DM组(P<0.05)。GLA-H组和MET组大鼠肝脏TG水平显著低于GLA-L组(P<0.05)。GLA-H组和MET组大鼠肝脏TC水平和TG水平比较差异均无统计学意义(P>0.05)。与CON组相比,DM组大鼠血清ALT和AST水平显著升高(P<0.05);与DM组相比,GLA-L组和GLA-H组大鼠ALT和AST水平均明显降低(P<0.05)。CON组与CON-GLA组大鼠肝脏PC、G6Pase、IDH2和CS蛋白相对表达量比较差异无统计学意义(P>0.05)。DM组、GLA-L组、GLA-H组和MET组大鼠肝脏组织中PC蛋白和G6Pase蛋白相对表达量显著高于CON组与CON-GLA组,IDH2和CS蛋白相对表达量显著低于CON组与CON-GLA组(P<0.05)。GLA-L组与DM组大鼠肝脏组织中PC和G6Pase蛋白相对表达量比较差异无统计学意义(P>0.05);GLA-H组和MET组大鼠肝脏组织中PC蛋白和G6Pase蛋白相对表达量显著低于DM组与GLA-L组(P<0.05)。GLA-H组与MET组大鼠肝脏组织中PC蛋白和G6Pase蛋白相对表达量比较差异无统计学意义(P>0.05)。GLA-L组、GLA-H组和MET组大鼠IDH2和CS蛋白相对表达量显著高于DM组,GLA-H组和MET组大鼠IDH2和CS蛋白相对表达量显著高于GLA-L组(P<0.05)。GLA-H组与MET组大鼠IDH2和CS蛋白相对表达量比较差异无统计学意义(P>0.05)。结论GLA可通过干预糖代谢缓解T2DM症状,降低血糖,并恢复大鼠体质量。
Objective To investigate the mitigative effect of glabridin(GLA)against type 2 diabetes mellitus(T2DM)and its mechanisms.Methods Thirty-six clean Sprague-Dawley(SD)rats were randomly divided into a control(CON)group,a GLA control(CON-GLA)group,a diabetes mellitus(DM)group,a low dose GLA(GLA-L)group,a high dose GLA(GLA-H)group and a metformin group(MET).The rats in CON and CON-GLA groups were given normal feeding.The rats in DM,GLA-L,GLA-H and MET groups were fed with a high-sugar and high-fat diet for 3 weeks to prepare T2DM rats.Then,while being fed with their respective diets,rats in the CON,CON-GLA,DM,GLA-L,GLA-H,and MET groups were given an intraperitoneal injection of 0.5 mL dimethyl sulfoxide(DMSO),0.5 mL GLA(20 mg·kg^(-1)),0.5 mL GLA(40 mg·kg^(-1))and 150 mg·kg^(-1)MET every two days for consecutive 28 days,respectively.The body weight(BW)and fasting blood glucose(FBG)were measured on Days 0,7,14,21 and 28 of administration after fasting for 10 h.Moreover,the oral glucose tolerance test(OGTT)was performed 1 day after the final administration,and the glucose area under the curve(AUC)was calculated.Fasting insulin(FINS)and glycated hemoglobin A1c(HbA1c)levels were measured by enzyme linked immunosorbent assay.Homeostatic model assessment for insulin resistance(HOMA-IR)and homeostatic model assessment forβ-cell function(HOMA-β)were calculated.At the same time,the serum levels of alanine amino-transferase(AST)and aspartate amino-transferase(ALT)were detected by a microplate reader.Fat deposition in the rat liver was observed by oil red O staining,and the liver triglyceride(TG)and total cholesterol(TC)content was detected.The expression of key rate-limiting enzymes pyruvate carboxylase(PC),glucose-6-phosphatase(G6Pase),citrate synthase(CS)and isocitrate dehydrogenase 2(IDH2)in different pathways of liver glucose metabolism was detected by Western blot assay.Results The BW of rats in DM,GLA-L,GLA-H and MET groups was significantly lower than that in the CON group on Days 0,7,14,21 and 28 of administration(P<0.05).The BW of rats in GLA-L,GLA-H and MET groups was significantly higher than that in the DM group and the BW of rats in GLA-H and MET groups was significantly higher than that in the GLA-L group on Days 14,21 and 28 of administration(P<0.05).The FBG levels of rats in DM and GLA-L groups were significantly higher than those in the CON and CON-GLA groups on Days 0,7,14,21 and 28 of administration(P<0.05).The FBG levels of rats in GLA-H and MET groups were significantly higher than those in CON and CON-GLA groups on Days 0,7,and 14 of administration(P<0.05).The FBG levels of rats in GLA-L,GLA-H and MET groups were significantly lower than those in the DM group on Days 7,14,21 and 28 of administration(P<0.05).The FBG levels of rats in GLA-H and MET groups were significantly lower than those in the GLA-L group on Days 21 and 28 of administration(P<0.05).Compared with those in the DM group,the blood glucose levels of rats in CON,CON-GLA,GLA-L,GLA-H and MET groups were significantly decreased at 30,60 and 120 min(P<0.05).In the OGTT testing,the blood glucose levels of rats in the GLA-L group decreased with time,and they were significantly higher than those in the CON group at 30,60 and 120 min(P<0.05).The blood glucose levels of rats in GLA-H and MET groups showed a similar change trend with those in the GLA-L group.The blood glucose levels of rats in GLA-H and MET groups were significantly higher than those in the CON group at 30 and 60 min and significantly lower than those in the GLA-L group at 120 min(P<0.05).The blood glucose levels of rats in GLA-H and MET groups were not significantly different from those in the CON group(P>0.05).There was no significant difference in blood glucose levels between GLA-H and MET groups at all test time points in the OGTT test(P>0.05).The glucose AUCs of rats in DM,GLA-L and GLA-H groups were significantly higher than those in the CON and CON-GLA groups(P<0.05).The glucose AUCs of rats in GLA-L,GLA-H and MET groups were significantly lower than those in the DM group(P<0.05).The glucose AUCs of rats in GLA-H and MET groups were significantly lower than those in the GLA-L group(P<0.05).There was no significant difference in glucose AUC between GLA-H and MET groups(P>0.05).The HbA1c levels of rats in the CON,CON-GLA,GLA-L,GLA-H and MET groups were significantly lower than those in the DM group(P<0.05).The HbA1c levels of rats in GLA-H and MET groups were significantly lower than those in the GLA-L group(P<0.05).The FINS level in the GLA-L group was significantly higher than that in the CON and CON-GLA groups(P<0.05).There was no significant difference in FINS and HOMA-IR levels between the GLA-H and CON groups(P>0.05).Compared with those in the DM group,FINS and HOMA-IR levels were significantly decreased while HOMA-βlevels were significantly increased in the CON,CON-GLA,GLA-L,GLA-H and MET groups(P<0.05).There was no significant difference in HOMA-βlevels among the CON,CON-GLA,GLA-L,GLA-H and MET groups(P>0.05).Oil red O staining results showed that there was little fat deposited in the liver of rats in CON and CON-GLA groups,but there was a large amount of fat deposited in the liver of rats in the DM group.The fat levels decreased gradually in the GLA-L and GLA-H groups,but they were still higher than those in the CON and CON-GLA groups.There was no significant difference in liver TC and TG levels between the CON and CON-GLA groups(P>0.05).The levels of TC in liver of rats in the DM,GLA-L,GLA-H and MET groups were significantly lower than those in the CON and CON-GLA groups(P<0.05).The levels of TG in liver of rats in the GLA-H and MET groups were not significantly different from those in the CON and CON-GLA groups(P>0.05).The levels of TG in liver of rats in the DM and GLA-L groups were significantly higher than those in the CON and CON-GLA groups(P<0.05).The levels of TC and TG in liver of rats in the GLA-L,GLA-H and MET groups were significantly lower than those in the DM group(P<0.05).The levels of TG in liver of rats in the GLA-H and MET groups were significantly lower than those in the GLA-L group(P<0.05).There was no significant difference in liver TC and TG levels between the GLA-H and MET groups(P>0.05).Compared with those in the CON group,the serum levels of ALT and AST in the DM group were significantly increased(P<0.05).Compared with those in the DM group,the serum levels of ALT and AST levels in GLA-L and GLA-H groups were significantly decreased(P<0.05).There was no significant difference in the liver expression levels of PC,G6Pase,IDH2 and CS proteins between the CON and CON-GLA groups(P>0.05).Compared with CON and CON-GLA groups,DM,GLA-L,GLA-H and MET groups showed significantly higher expression levels of PC and G6Pase proteins and significantly lower expression levels of IDH2 and CS in the liver(P<0.05).There was no significant difference in the liver expression levels of PC and G6Pase proteins between the GLA-L and DM groups(P>0.05).The liver expression levels of PC and G6Pase proteins in GLA-H and MET groups were significantly lower than those in DM and GLA-L groups(P<0.05).There was no significant difference in the liver expression levels of PC and G6Pase proteins between GLA-H and MET groups(P>0.05).The liver expression levels of IDH2 and CS proteins in GLA-L,GLA-H and MET groups were significantly higher than those in the DM group,and the liver expression levels of IDH2 and CS proteins in GLA-H and MET groups were significantly higher than those in the GLA-L group(P<0.05).There was no significant difference in the liver expression levels of IDH2 and CS proteins between GLA-H and MET groups(P>0.05).Conclusion GLA can alleviate T2DM,reduce blood glucose and restore rat BW through interfering with glucose metabolism.
作者
冯亚娟
冯亚静
周建华
王旭东
孙鹏
FENG Yajuan;FENG Yajing;ZHOU Jianhua;WANG Xudong;SUN Peng(Department of Pharmacy,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,Henan Province,China;Department of Infection Management,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,Henan Province,China;Department of Pharmacy,the Second Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450002,Henan Province,China)
出处
《新乡医学院学报》
2025年第4期266-276,共11页
Journal of Xinxiang Medical University
基金
河南省高校科技创新团队支持计划(编号:14IRTSTHN017)。
