利拉鲁肽对糖尿病大鼠肾组织炎症因子表达的影响
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摘要
目的观察利拉鲁肽对糖尿病肾病大鼠肾组织Toll样受体(TLR) 4/髓样细胞分化因子(MyD) 88信号转导通路中炎症因子的影响。方法健康SD雄性大鼠68只,随机分为正常对照组、模型组、利拉鲁肽组、二甲双胍组,每组17只。采用高脂高糖饮食饲养联合腹腔注射少量链脲佐菌素(STZ)法制备大鼠糖尿病肾病模型。利拉鲁肽组按0. 12 mg·kg~(-1)·d~(-1)的剂量皮下注射给药,二甲双胍组按140 mg·kg~(-1)·d~(-1)的剂量灌胃给药,正常对照组及模型组均给予等量溶媒,实验期间各组大鼠均无降糖干预,饲养8 w。检测尿蛋白、尿糖及血生化,苏木素-伊红(HE)染色观察大鼠肾组织形态学改变,免疫组化检测TLR4/MyD88蛋白表达,酶联免疫吸附试验(ELISA)检测大鼠肾组织核因子(NB)-κB、肿瘤坏死因子(TNF)-α及白细胞介素(IL)-6含量。结果与正常对照组相比,模型组、利拉鲁肽组、二甲双胍组24 h尿蛋白含量、24 h尿糖、血肌酐、尿素氮及血糖水平明显升高,TLR4/MyD88蛋白表达明显增多,NB-κB、TNF-α、IL-6含量明显上升(均P<0. 05)。与模型组相比,利拉鲁肽组、二甲双胍组24 h尿蛋白含量、24 h尿糖、血肌酐、尿素氮及血糖明显下降,TLR4/MyD88蛋白表达明显降低(P<0. 05),NB-κB、TNF-α、IL-6表达明显下降(P<0. 05)。结论利拉鲁肽能对糖尿病大鼠肾脏产生保护,其机制可能与其通过调控TLR4/MyD88信号通路抑制炎症因子NB-κB、TNF-α、IL-6的表达有关。
Objective To observe the effect of liraglutide on TLR4/MyD88 path renal tissue of diabetic rats and provide a way to treat diabetic nephropathy.Methods 68 SD Wistar rats were randomly divided into normal control,model and liraglutide,metformin groups,17 in each group. Model of diabetic nephropathy was made by combined high fat and sugar diet feeding with intraperitoneal injection of a small dose of STZ. Rats were given 0.12 mg·kg-1·d-1 liraglutide subcutaneously in liraglutide group,140 mg·kg-1·d-1 metformin lavage metformin group,the same amount of solvent in normal control,model groups. All rats in the experimental period should be fed for 8 weeks without insulin. 24 hours urine protein,24 hours urine sugar,serum creatinine,blood urea nitrogen and blood sugar levels were measured and under optical microscope after HE staining in the rat kidney tissue morphology change was observed,TLR4/MyD88 protein expression was detected by immunohistochemistry,NF-κB,TNF-α and IL-6 content in rat kidney were measured by ELISA. Results Compared with normal control group,24 hours urinary protein,24 hours urine sugar,serum creatinine,blood urea nitrogen were elevated,TLR4/MyD88 protein expression were increased,NB-κB,TNF-α,IL-6 expression were decreased( all P < 0. 05) in model,liraglutide and metformin groups. Compared with model group,24 hours urinary protein,24 hours urine sugar,serum creatinine,blood urea nitrogen and blood sugar drops,TLR4/MyD88 protein were decreased,NB-κB,TNF-α,IL-6 expression were increased( all P< 0.05) in liraglutide and metformin groups.Conclusions Liraglutide could protect the kidneys of diabetic rats,the mechanism may be related with regulation of TLR4/MyD88 signaling pathway,inhibiting inflammatory cytokines NB-κB,TNF-α and IL-6.
Objective To observe the effect of liraglutide on TLR4/MyD88 path renal tissue of diabetic rats and provide a way to treat diabetic nephropathy.Methods 68 SD Wistar rats were randomly divided into normal control,model and liraglutide,metformin groups,17 in each group. Model of diabetic nephropathy was made by combined high fat and sugar diet feeding with intraperitoneal injection of a small dose of STZ. Rats were given 0.12 mg·kg-1·d-1 liraglutide subcutaneously in liraglutide group,140 mg·kg-1·d-1 metformin lavage metformin group,the same amount of solvent in normal control,model groups. All rats in the experimental period should be fed for 8 weeks without insulin. 24 hours urine protein,24 hours urine sugar,serum creatinine,blood urea nitrogen and blood sugar levels were measured and under optical microscope after HE staining in the rat kidney tissue morphology change was observed,TLR4/MyD88 protein expression was detected by immunohistochemistry,NF-κB,TNF-α and IL-6 content in rat kidney were measured by ELISA. Results Compared with normal control group,24 hours urinary protein,24 hours urine sugar,serum creatinine,blood urea nitrogen were elevated,TLR4/MyD88 protein expression were increased,NB-κB,TNF-α,IL-6 expression were decreased( all P < 0. 05) in model,liraglutide and metformin groups. Compared with model group,24 hours urinary protein,24 hours urine sugar,serum creatinine,blood urea nitrogen and blood sugar drops,TLR4/MyD88 protein were decreased,NB-κB,TNF-α,IL-6 expression were increased( all P< 0.05) in liraglutide and metformin groups.Conclusions Liraglutide could protect the kidneys of diabetic rats,the mechanism may be related with regulation of TLR4/MyD88 signaling pathway,inhibiting inflammatory cytokines NB-κB,TNF-α and IL-6.
引文
1王玉华,蔡春友,李卫东,等.全外显子组测序发现FUT6基因在1例糖尿病肾病中的终止突变[J].天津医科大学学报,2015; 21(1):18-21.
2 闫寒,马博清,付彩雯.糖尿病肾病发病机制研究进展[J].中国老年学杂志,2015; 35(20):5973-5.
3 Feng Y,Yang S,Ma Y,et al.Role of Toll-like receptors in diabetic renal lesions in a miniature pig model[J].Sci Adv,2015; 1(15):e1400183.
4 张晶晶,魏蕊,杨进,等.利拉鲁肽延缓2型糖尿病进展的潜在机制:中国学者的研究证据[J].中国糖尿病杂志,2015; 23(12):1057-68.
5 江思瑜,孙霓,李佳航,等.2型糖尿病大鼠肾组织NF-κB水平的变化[J].中国老年学杂志,2018; 38(14):3439-41.
6 Lv JL,Chen QK,Shao Y,et al. Cross-talk between angiotensin-Ⅱand toll-like receptor 4 triggers a synergetic inflammatory response in rat mesangial cells under high glucose conditions[J]. Biochem Biophys Res Commun,2015; 459(2):264-9.
7 危正南,李涛,张庆红,等. MIF、TLR4、TNF-α水平在糖尿病肾病患者中的变化及其临床意义[J].疑难病杂志,2016; 15(2):165-8.
8 Liu P,Li F,Qiu MC,et al.Expression and cellular distribution of TLR4,My D88,and NF-κB in diabetic renal tubulointerstitial fibrosis,in vitro and in vivo[J].Diabetes Res Clin Practice,2014; 105(2):206-16.
9 李曼丽,甘华.高糖对肾小管上皮细胞Toll样受体4表达及其信号通路的影响[J].重庆医科大学学报,2010; 35(6):857-9.
10 常微微,贺连平,姚应水. Toll样受体4与糖尿病肾病的发病机制相关性的研究进展[J].中国临床药理学杂志,2015; 31(11):1066-8.
11 熊梅梅,吕柳青,肖红波,等.高糖和血管紧张素Ⅱ对人肾小管上皮细胞Toll样受体4信号表达及炎性和纤维化因子的影响[J].中华肾脏病杂志,2016; 32(1):43-9.
12 孙凤杰,董文斌. TLR4/My D88信号通路与肾脏疾病相关性的研究进展[J].泸州医学院学报,2010; 33(1):100-2.
13 章烈,李园,周总光.髓样分化因子88的研究进展:把握TLR4信号通路中的瓶颈[J].中国普外基础与临床杂,2013; 20(6):685-90.
14 朱雪坤,杨日,孟艳秋. Toll样受体4配体的研究进展[J].中国药理学与毒理学杂志,2016; 30(3):78-85.
15 吴燕燕,王易. Toll样受体信号通路中My D88的研究进展[J].免疫学杂志,2012; 28(3):262-5.
16 刘璠,郭玉卿,周慧敏,等.利拉鲁肽对2型糖尿病患者血清因子的影响[J].现代中西医结合杂志,2015; 24(11):1147-9.
17 赵春云,郭洪涛,代红沙,等.利拉鲁肽治疗早期2型糖尿病肾病疗效观察[J].山东医药,2014; 54(26):47-9.
18 刘璠,张趁儒,周慧敏,等.人GLP-1类似物对2型糖尿病患者血清IL-6、TNF-α、CRP及脂联素的影响[J].中国现代医学杂志,2014; 24(53):27-30.
19 崔智慧,崔岩,孙高洁,等.利拉鲁肽对小鼠脂肪细胞脂肪因子、炎症因子及其信号通路的影响[J].郑州大学学报(医学版),2015;(4):519-22.
20 李里,卢松,甘华.利拉鲁肽对糖尿病大鼠肾脏巨噬细胞浸润的影响[J].第三军医大学学报,2014; 36(18):1909-13.
21 Sai J,Zhou L,Bai,et al.Liraglutide ameliorates renal injury in streptozotocin induced diabetic rats by activating endothelial nitric oxide synthase activity via the downregulation of the nuclear factorκB pathway[J].Mol Med Rep,2014; 10(5):2587-94.
2 闫寒,马博清,付彩雯.糖尿病肾病发病机制研究进展[J].中国老年学杂志,2015; 35(20):5973-5.
3 Feng Y,Yang S,Ma Y,et al.Role of Toll-like receptors in diabetic renal lesions in a miniature pig model[J].Sci Adv,2015; 1(15):e1400183.
4 张晶晶,魏蕊,杨进,等.利拉鲁肽延缓2型糖尿病进展的潜在机制:中国学者的研究证据[J].中国糖尿病杂志,2015; 23(12):1057-68.
5 江思瑜,孙霓,李佳航,等.2型糖尿病大鼠肾组织NF-κB水平的变化[J].中国老年学杂志,2018; 38(14):3439-41.
6 Lv JL,Chen QK,Shao Y,et al. Cross-talk between angiotensin-Ⅱand toll-like receptor 4 triggers a synergetic inflammatory response in rat mesangial cells under high glucose conditions[J]. Biochem Biophys Res Commun,2015; 459(2):264-9.
7 危正南,李涛,张庆红,等. MIF、TLR4、TNF-α水平在糖尿病肾病患者中的变化及其临床意义[J].疑难病杂志,2016; 15(2):165-8.
8 Liu P,Li F,Qiu MC,et al.Expression and cellular distribution of TLR4,My D88,and NF-κB in diabetic renal tubulointerstitial fibrosis,in vitro and in vivo[J].Diabetes Res Clin Practice,2014; 105(2):206-16.
9 李曼丽,甘华.高糖对肾小管上皮细胞Toll样受体4表达及其信号通路的影响[J].重庆医科大学学报,2010; 35(6):857-9.
10 常微微,贺连平,姚应水. Toll样受体4与糖尿病肾病的发病机制相关性的研究进展[J].中国临床药理学杂志,2015; 31(11):1066-8.
11 熊梅梅,吕柳青,肖红波,等.高糖和血管紧张素Ⅱ对人肾小管上皮细胞Toll样受体4信号表达及炎性和纤维化因子的影响[J].中华肾脏病杂志,2016; 32(1):43-9.
12 孙凤杰,董文斌. TLR4/My D88信号通路与肾脏疾病相关性的研究进展[J].泸州医学院学报,2010; 33(1):100-2.
13 章烈,李园,周总光.髓样分化因子88的研究进展:把握TLR4信号通路中的瓶颈[J].中国普外基础与临床杂,2013; 20(6):685-90.
14 朱雪坤,杨日,孟艳秋. Toll样受体4配体的研究进展[J].中国药理学与毒理学杂志,2016; 30(3):78-85.
15 吴燕燕,王易. Toll样受体信号通路中My D88的研究进展[J].免疫学杂志,2012; 28(3):262-5.
16 刘璠,郭玉卿,周慧敏,等.利拉鲁肽对2型糖尿病患者血清因子的影响[J].现代中西医结合杂志,2015; 24(11):1147-9.
17 赵春云,郭洪涛,代红沙,等.利拉鲁肽治疗早期2型糖尿病肾病疗效观察[J].山东医药,2014; 54(26):47-9.
18 刘璠,张趁儒,周慧敏,等.人GLP-1类似物对2型糖尿病患者血清IL-6、TNF-α、CRP及脂联素的影响[J].中国现代医学杂志,2014; 24(53):27-30.
19 崔智慧,崔岩,孙高洁,等.利拉鲁肽对小鼠脂肪细胞脂肪因子、炎症因子及其信号通路的影响[J].郑州大学学报(医学版),2015;(4):519-22.
20 李里,卢松,甘华.利拉鲁肽对糖尿病大鼠肾脏巨噬细胞浸润的影响[J].第三军医大学学报,2014; 36(18):1909-13.
21 Sai J,Zhou L,Bai,et al.Liraglutide ameliorates renal injury in streptozotocin induced diabetic rats by activating endothelial nitric oxide synthase activity via the downregulation of the nuclear factorκB pathway[J].Mol Med Rep,2014; 10(5):2587-94.