降糖三黄片对高糖高脂饮食大鼠胰岛β细胞去分化的作用

Effect of Jiangtang Sanhuang Tablets on Islet β-cells dedifferentiation in Rats Induced by High-Glucose and High-Fat Diet

【目的】探讨降糖三黄片对高糖高脂饮食致大鼠胰岛β细胞去分化的作用。【方法】将30只雄性SD大鼠随机分为正常组、模型组和中药组,每组10只,分别给予普通饮食、高糖高脂饮食、高糖高脂饮食并每天灌胃中成药降糖三黄片(675 mg·kg^-1·d^-1)。连续干预180 d后,检测血清空腹血糖(FBG)、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、空腹胰岛素(FINS),采用苏木素—伊红染色评估胰腺组织病理学变化,免疫组织化学法检测胰腺组织胰岛素(Insulin)、胰十二指肠同源盒因子(Pdx1)、肌腱膜纤维肉瘤癌基因同源物A蛋白(MafA)、叉头状转录因子(FOXO1)蛋白的表达。基于高通量蛋白芯片技术筛选血清差异表达细胞因子。【结果】与正常组比较,模型组大鼠FBG、TC、TG、LDL-C、HOMA-IR水平升高,FINS、HOMA-β、HDL-C水平下降,胰岛β细胞Insulin、Pdx1、MafA蛋白表达下调,FOXO1蛋白表达上调(均P<0.05)。基于高通量的蛋白芯片检测及分析,筛选出正常组和模型组有显著性差异的11种细胞因子:金属蛋白酶组织抑制物1(TIMP-1)、细胞间黏附分子1(ICAM-1)、Gas 1、肿瘤坏死因子样凋亡弱诱导因子受体(TWEAK R)、神经纤毛蛋白2(Neuropilin-2)、LIX、白细胞介素13(IL-13)、激活素A(Activin A)、血浆嗜酸性粒细胞趋化因子(Eotaxin)、半乳凝素3(Galectin-3)、Decorin(P<0.05或P<0.01)。与模型组比较,中药组大鼠FBG、TC、TG、LDL-C、HOMA-IR水平下降,FINS、HOMA-β、HDL-C水平升高,胰岛β细胞Insulin、Pdx1、MafA蛋白表达上调,FOXO1蛋白表达下调(均P<0.05)。基于高通量的蛋白芯片检测及分析,筛选出模型组和中药组有显著性差异的4个细胞因子:Galectin-3、TIMP-1、Activin A、Eotaxin(P<0.05或P<0.01)。【结论】降糖三黄片具有调控FOXO1/Pdx1通路从而延缓"糖脂毒性"作用下的β细胞去分化的作用,并与降低血清细胞因子Galectin-3、TIMP-1、Activin A、Eotaxin含量密切相关。

Objective To explore the effect of Jiangtang Sanhuang Tablets on islet β-cells dedifferentiation in ratsinduced by high-glucose and high-fat diet. Methods Thirty male SD rats were randomly divided into normalgroup,model group and Chinese medicine group,10 rats in each group,and were respectively correspondinglygiven ordinary diet,high-glucose and high-fat diet,high-glucose and high-fat diet plus everyday intragastric administration of Chinese patent drug Jiangtang Sanhuang Tablets(675 mg · kg^-1· d^-1)for 180 days. Afterintervention,fasting blood glucose(FBG),triglyceride(TG),total cholesterol(TC),high density lipoproteinc holesterol(HDL-C),low density lipoprotein cholesterol(LDL-C)and fasting insulin(FINS)were measured.The pathological features of pancreatic tissue was evaluated by hematoxylin-eosin(HE)staining. The protein expressionof Insulin, pancreatic and duodenal homeobox 1(Pdx1), v-maf muscle oaponeurotic fibrosarcoma oncogene homologue A(Maf A), and forkhead box O1(FOXO1)in pancreas were detected by immunohistochemistry.G-Series Rat Cytokine Antibody Array 67 was used to profile the variation of proteins in serum. Results Comparedwith the normal group,the levels of FBG,TG,TC,LDL-C and HOMA-IR were increased,FINS,HOMA-β,HDL-C were decreased, protein expression levels of Insulin, Pdx1 and Maf A in islet β-cells were down-regulated, and protein expression level of FOXO1 was up-regulated(all P < 0.05). G-Series Rat CytokineAntibody Array 67 results showed that there were significant differences in 11 cytokines including TIMP-1,ICAM-1,Gas 1,TNF-like weak inducer of apoptosis receptor(TWEAK R),Neuropilin-2,LIX,interleukin 13(IL-13),Activin A, Eotaxin, Galectin-3, Decorin between the two groups(P<0.05 or P<0.01). Compared with themodel group,the levels of FBG,TG,TC,LDL-C and HOMA-IR were decreased,FINS,HOMA-β,HDL-C were increased,protein expression levels of Insulin,Pdx1 and Maf A in the islet β-cells were up-regulated,protein expression level of FOXO1 was down-regulated(all P < 0.05). G-Series Rat Cytokine Antibody Array 67 results showed that there were significant differences in 4 cytokines including Galectin-3,TIMP-1,Activin A,and Eotaxin between the two groups(P<0.05 or P<0.01). Conclusion Jiangtang Sanhuang Tablets have effectson reversing the β-cell dedifferentiation induced by high-glucose and high-fat diet,which is related with loweringthe serum levels of Galectin-3,TIMP-1,Activin A,and Eotaxin.