利拉鲁肽对非酒精性脂肪肝大鼠肝脏组织中胰岛素JNK1信号通路的影响
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摘要
目的探究利拉鲁肽对非酒精性脂肪肝(NAFLD)大鼠肝脏组织中胰岛素JNK1信号通路的影响。方法选取40只6周龄SPF级大鼠,采用高脂饮食喂养12周建立NAFLD大鼠模型。将NAFLD大鼠随机分为空白对照组(control)、模型组(model)、低剂量利拉鲁肽组(low lira)和高剂量利拉鲁肽组(high lira)。利拉鲁肽干预18 d后,测量大鼠体质量及肝指数变化;全自动生化分析仪测定大鼠血清丙氨酸氨基转移酶(ALT)、空腹血糖(FBG)、甘油三酯(TG)、总胆固醇(TC)和血清胰岛素(FINS)变化;酶联免疫吸附法测定大鼠肝组织中肿瘤坏死因子(TNF-α)、超氧化物歧化酶(SOD)、丙二醛(MAD)及游离脂肪酸(FFAs)含量; Western blot检测大鼠肝脏组织中胰岛素受体(IR)、磷酸化胰岛素受体底物1(p-IRS1)、C-Jun氨基端激酶1(JNK1)及磷酸化C-Jun氨基端激酶1(p-JNK1)表达情况; HE染色观察大鼠肝脏组织病理学变化。结果与对照组相比,模型组大鼠体质量,肝指数,血清ALT、TG、TC、FINS、TNF-α、MAD、FFAs含量,p-IRS1、JNK1、p-JNK1蛋白表达量显著升高,SOD含量显著降低,差异有统计学意义(P <0. 01);血清FBG含量和IR蛋白表达量无显著变化(P> 0. 05);相比模型组,利拉鲁肽组大鼠体质量,肝指数,血清ALT、TG、TC、FINS、TNF-α、MAD、FFAs含量,p-IRS1、JNK1、p-JNK1蛋白表达量显著降低,且高剂量组显著低于低剂量组(P <0. 01); SOD含量显著升高,且高剂量组显著高于低剂量组(P <0. 01);血清FBG含量和IR蛋白表达量无显著变化(P> 0. 05)。结论利拉鲁肽可以缓解大鼠脂肪肝的发展,其作用机制可能与抑制JNK1信号通路及JNK1磷酸化相关,且在一定范围内呈浓度依赖性。
Objective To explore the effects of liraglutide on insulin JNK1 signaling pathway in liver tissues of rats with nonalcoholic fatty liver disease. Methods Forty 6-week-old SPF rats were fed with a high-fat diet for 12 weeks to establish the NAFLD rat model. NAFLD rats were randomly divided into blank control group( control), model group( model), low dose liraglutide group( low lira) and high dose liraglutide group( high lira). After 18 days of intervention with liraglutide, the changes of body weight and liver indexes were measured. The serum alanine aminotransferase( ALT), fasting blood glucose( FBG), triglyceride( TG), total cholesterol( TC) and serum insulin( FINS) were measured by automatic biochemical analyzer. The levels of tumor necrosis factor( TNF-α), superoxide dismutase( SOD), malondialdehyde( MAD) and free fatty acid( FFAs) in rat liver tissues were detected by enzyme-linked immunosorbent assay. Western blot was used to detect the expression levels of insulin receptor( IR), phosphorylated insulin receptor substrate 1( p-IRS1), C-Jun N-terminal kinase 1( JNK1) and phosphorylation C-Jun N-terminal kinase 1( p-JNK1) in rat liver tissues. The pathological changes in rat liver tissues were observed by HE staining. Results Compared with control group,the body weight, liver indexes, levels of serum ALT, TG, TC, FINS, TNF-α, MAD and FFAs, and protein expression levels of p-IRS1, JNK1 and p-JNK1 in model group were increased significantly while the SOD level was decreased significantly( P < 0. 01).There were no significant changes in the serum FBG level and IR protein expression level( P > 0. 05). Compared with model group, the body weight, liver indexes, levels of serum ALT, TG, TC, FINS, TNF-α, MAD and FFAs, and protein expression levels of p-IRS1,JNK1 and p-JNK1 in liraglutide groups were decreased significantly, and the indexes in high dose group were significantly lower than those in low dose group( P < 0. 01). The SOD level was increased significantly, and the level in high dose group was significantly higher than that in low dose group( P < 0. 01), and there were no significant changes in the serum FBG level and IR protein expression level( P > 0. 05). Conclusion Liraglutide can alleviate the development of fatty liver in rats. Its mechanism may be related to the inhibition of JNK1 signaling pathway and JNK1 phosphorylation in a concentration-dependent manner.
Objective To explore the effects of liraglutide on insulin JNK1 signaling pathway in liver tissues of rats with nonalcoholic fatty liver disease. Methods Forty 6-week-old SPF rats were fed with a high-fat diet for 12 weeks to establish the NAFLD rat model. NAFLD rats were randomly divided into blank control group( control), model group( model), low dose liraglutide group( low lira) and high dose liraglutide group( high lira). After 18 days of intervention with liraglutide, the changes of body weight and liver indexes were measured. The serum alanine aminotransferase( ALT), fasting blood glucose( FBG), triglyceride( TG), total cholesterol( TC) and serum insulin( FINS) were measured by automatic biochemical analyzer. The levels of tumor necrosis factor( TNF-α), superoxide dismutase( SOD), malondialdehyde( MAD) and free fatty acid( FFAs) in rat liver tissues were detected by enzyme-linked immunosorbent assay. Western blot was used to detect the expression levels of insulin receptor( IR), phosphorylated insulin receptor substrate 1( p-IRS1), C-Jun N-terminal kinase 1( JNK1) and phosphorylation C-Jun N-terminal kinase 1( p-JNK1) in rat liver tissues. The pathological changes in rat liver tissues were observed by HE staining. Results Compared with control group,the body weight, liver indexes, levels of serum ALT, TG, TC, FINS, TNF-α, MAD and FFAs, and protein expression levels of p-IRS1, JNK1 and p-JNK1 in model group were increased significantly while the SOD level was decreased significantly( P < 0. 01).There were no significant changes in the serum FBG level and IR protein expression level( P > 0. 05). Compared with model group, the body weight, liver indexes, levels of serum ALT, TG, TC, FINS, TNF-α, MAD and FFAs, and protein expression levels of p-IRS1,JNK1 and p-JNK1 in liraglutide groups were decreased significantly, and the indexes in high dose group were significantly lower than those in low dose group( P < 0. 01). The SOD level was increased significantly, and the level in high dose group was significantly higher than that in low dose group( P < 0. 01), and there were no significant changes in the serum FBG level and IR protein expression level( P > 0. 05). Conclusion Liraglutide can alleviate the development of fatty liver in rats. Its mechanism may be related to the inhibition of JNK1 signaling pathway and JNK1 phosphorylation in a concentration-dependent manner.
引文
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[13]李娜,黄鑫源,李敏.25-(OH)D3、ADPN水平及IR对NAFLD的临床诊断价值[J].热带医学杂志,2017,17(4):491-494.
[14]霍琴琴,钟兴,杜益君,等.利拉鲁肽对非酒精性脂肪肝大鼠模型疗效评价[J].安徽医科大学学报,2017,52(7):967-972.
[15]刘丽波,张梦梦,方权明,等.非酒精性脂肪肝患者口服葡萄糖后血浆胰高血糖素样肽和葡萄糖依赖性促胰岛素多肽水平的变化[J].中国现代应用药学,2016,33(3):368-371.
[16]帖彦清,张明明,李静,等.不同类型脂肪肝患者血清HMGB1水平与氧化应激及炎性反应的关系[J].河北医药,2018,40(6):833-836.
[17]Chaure J,Serrano C,Fernández-Parra R,et al.On Studying the Interaction Between Different Stent Models and Rabbit Tracheal Tissue:Numerical,Endoscopic and Histological Comparison[J].Ann Biomed Eng.2016,44(2):368-381.
[18]庞晓宁,刘彦君.利拉鲁肽、西他列汀和吡格列酮治疗NAFLD合并2型糖尿病患者疗效初步比较研究[J].实用肝脏病杂志,2017,20(5):604-605.
[2]Kaswala DH,Lai M,Afdhal NH.Fibrosis Assessment in Nonalcoholic Fatty Liver Disease(NAFLD)in 2016[J].Dig Dis Sci,2016,61(5):1356-1364.
[3]Buzzetti E,Pinzani M,Tsochatzis EA.The multiple-hit pathogenesis of non-alcoholic fatty liver disease(NAFLD)[J].Metabolism,2016,65(8):1038-1048.
[4]Smith BK,Marcinko K,Desjardins EM,et al.Treatment of nonalcoholic fatty liver disease:role of AMPK[J].Am J Physiol Endocrinol Metab,2016,1;311(4):E730-E740.
[5]Mann JFE,Orsted DD,Brown-Frandsen K,et al.Liraglutide and Renal Outcomes in Type 2 Diabetes[J].N Engl J Med,2016,377(9):839-848.
[6]Luo G,Liu H,Lu H.Glucagon-like peptide-1(GLP-1)receptor agonists:potential to reduce fracture risk in diabetic patients?[J].Br J Clin Pharmacol,2016,81(1):78-88.
[7]杨颖博,田小燕,黄一茜,等.利拉鲁肽联合胰岛素和格列吡嗪治疗亚甲减合并2型糖尿病老年患者的临床研究[J].中国药房,2017,28(14):1958-1961.
[8]朱晓宁,汪静,郑丁,等.非酒精性脂肪性肝病痰瘀互结证大鼠模型的构建及评价[J].中医杂志,2017,58(22):1953-1956.
[9]Vauzour D,Rodriguez-Ramiro I,Rushbrook S,et al.N-3 fatty acids combined with flavan-3-ols prevent steatosis and liver injury in a murine model of NAFLD[J].Biochim Biophys Acta Mol Basis Dis.,2018,1864(1):69-78.
[10]Qiu H,Yang JK,Chen C.Influence of insulin on growth hormone secretion,level and growth hormone signalling[J].Acta Physiologica Sinica,2017,69(5):541-556.
[11]Nishina A,Itagaki M,Sato D,et al.The Rosiglitazone-Like Effects of Vitexilactone,a Constituent from Vitex trifolia L.in 3T3-L1 Preadipocytes[J].Molecules,2017,22(11):E2030.
[12]石志平,杨昱,雷营,等.利拉鲁肽降低2型糖尿病伴非酒精性脂肪性肝病大鼠肝脏磷酸化c-Jun氨基末端激酶的蛋白表达[J].中国糖尿病杂志,2016,8(9):548-553.
[13]李娜,黄鑫源,李敏.25-(OH)D3、ADPN水平及IR对NAFLD的临床诊断价值[J].热带医学杂志,2017,17(4):491-494.
[14]霍琴琴,钟兴,杜益君,等.利拉鲁肽对非酒精性脂肪肝大鼠模型疗效评价[J].安徽医科大学学报,2017,52(7):967-972.
[15]刘丽波,张梦梦,方权明,等.非酒精性脂肪肝患者口服葡萄糖后血浆胰高血糖素样肽和葡萄糖依赖性促胰岛素多肽水平的变化[J].中国现代应用药学,2016,33(3):368-371.
[16]帖彦清,张明明,李静,等.不同类型脂肪肝患者血清HMGB1水平与氧化应激及炎性反应的关系[J].河北医药,2018,40(6):833-836.
[17]Chaure J,Serrano C,Fernández-Parra R,et al.On Studying the Interaction Between Different Stent Models and Rabbit Tracheal Tissue:Numerical,Endoscopic and Histological Comparison[J].Ann Biomed Eng.2016,44(2):368-381.
[18]庞晓宁,刘彦君.利拉鲁肽、西他列汀和吡格列酮治疗NAFLD合并2型糖尿病患者疗效初步比较研究[J].实用肝脏病杂志,2017,20(5):604-605.