黄芪对糖尿病肾病小鼠内质网应激中蛋白激酶R样内质网激酶通路的影响
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摘要
目的探讨黄芪对糖尿病肾病(DN)小鼠内质网应激中蛋白激酶R样内质网激酶(PERK)通路的影响。方法 KKAy小鼠和C57BL/6J小鼠分别给予高脂饲料和普通饲料,连续饲养2周。将建模成功的KKAy小鼠随机分为模型组和实验组,每组15只,15只C57BL/6J小鼠为对照组。实验组小鼠经腹腔注射黄芪注射液0. 003 mL·g~(-1),qd,连续干预4周;模型组和对照组均腹腔注射等量0. 9%NaCl。用全自动生化分析仪检测小鼠血清生化指标,苏木精-伊红(HE)染色法和蛋白免疫印迹法观察小鼠肾组织病理学形态及肾组织组织PERK通路相关蛋白表达。结果对照组,模型组,实验组体质量分别为(29. 20±1. 20),(48. 40±6. 90),(35. 10±2. 40) g,肾质量分别为(0. 16±0. 01),(0. 23±0. 01),(0. 20±0. 02) g,空腹血糖(FBG)分别为(5. 36±1. 25),(33. 08±5. 42),(23. 15±1. 05) mmol·L~(-1),肌酐(CREA)分别为(11. 69±0. 51),(10. 23±0. 28),(10. 95±0. 36)μmol·L~(-1),尿素(URE)分别为(8. 96±0. 84),(13. 52±0. 96),(10. 22±0. 69) mmol·L~(-1); 78kD-葡萄糖调节蛋白(GRP78)蛋白相对表达量分别为0. 23±0. 06,0. 79±0. 22,0. 53±0. 16; PERK蛋白相对表达量分别为0. 19±0. 04,0. 81±0. 16,0. 62±0. 19。实验组与模型组比较,差异均有统计学意义(均P <0. 01)。结论黄芪可降低DN小鼠体质量,改善肾组织损伤程度及肾功能,可能与下调PERK通路蛋白表达有关。
Objective To explore the effect of astragalus membranaceus on Protein kinase R like endoplasmic reticulum kinase( PERK) pathway in endoplasmic reticulum stress of diabetic nephropathy( DN) mice.Methods KKAymice and C57BL/6J mice were fed with high fat diet and common diet respectively for 2 weeks. The diabetic KKAymice were randomly divided into model group and test group,with 15 mice in each group. Fifteen 15 C57BL/6J mice were assigned to control group. The mice in test group were intraperitoneally injected with 0. 003 mL·g~(-1) of astragalus membranaceus. The mice in model group and normal group were intraperitoneally injected with the equal amount of 0. 9% NaCl once daily for 4 weeks. Serum biochemical indexes were detected by automatic biochemical analyzer. Hematoxylin-eosin( HE) staining and Western blot were used to observe the pathological morphology and the expression of PERK pathway protein in kidney tissue of mice. Results The body weight of control group,model group and test group were( 29. 20 ± 1. 20) g,( 48. 40 ± 6. 90) g,( 35. 10 ± 2. 40) g,and renal mass was( 0. 16 ± 0. 01) g,( 0. 23 ± 0. 01) g,( 0. 20 ± 0. 02) g,respectively; fasting blood glucose( FBG) were( 5. 36 ± 1. 25) mmol·L~(-1),( 33. 08 ± 5. 42) mmol·L~(-1),( 23. 15 ± 1. 05) mmol·L~(-1),respectively;creatinine( CREA) were( 11. 69 ± 0. 51) μmol·L~(-1),( 10. 23 ± 0. 28) μmol·L~(-1),( 10. 95 ± 0. 36) μmol·L~(-1),respectively; urinary albumin excretion rate( URE) were( 8. 96 ± 0. 84) μmol·L~(-1),( 13. 52 ± 0. 96) μmol·L~(-1),( 10. 22 ± 0. 69) μmol·L~(-1),respectively; the relative expression level of 78 kD-glucose regulated protein( GRP78)in normal,model and test groups were 0. 23 ± 0. 06,0. 79 ± 0. 22,0. 53 ± 0. 16,respectively; the relative expression level of PERK protein were 0. 19 ± 0. 04,0. 81 ± 0. 16,0. 62 ± 0. 19,respectively. There were statistically significant differences between the test group and the model group( P < 0. 05). Conclusion Astragalus membranaceus can reduce the body mass of DN mice,improve the degree of renal tissue damage and renal function,which may be related to the down-regulation of PERK pathway protein expression.
Objective To explore the effect of astragalus membranaceus on Protein kinase R like endoplasmic reticulum kinase( PERK) pathway in endoplasmic reticulum stress of diabetic nephropathy( DN) mice.Methods KKAymice and C57BL/6J mice were fed with high fat diet and common diet respectively for 2 weeks. The diabetic KKAymice were randomly divided into model group and test group,with 15 mice in each group. Fifteen 15 C57BL/6J mice were assigned to control group. The mice in test group were intraperitoneally injected with 0. 003 mL·g~(-1) of astragalus membranaceus. The mice in model group and normal group were intraperitoneally injected with the equal amount of 0. 9% NaCl once daily for 4 weeks. Serum biochemical indexes were detected by automatic biochemical analyzer. Hematoxylin-eosin( HE) staining and Western blot were used to observe the pathological morphology and the expression of PERK pathway protein in kidney tissue of mice. Results The body weight of control group,model group and test group were( 29. 20 ± 1. 20) g,( 48. 40 ± 6. 90) g,( 35. 10 ± 2. 40) g,and renal mass was( 0. 16 ± 0. 01) g,( 0. 23 ± 0. 01) g,( 0. 20 ± 0. 02) g,respectively; fasting blood glucose( FBG) were( 5. 36 ± 1. 25) mmol·L~(-1),( 33. 08 ± 5. 42) mmol·L~(-1),( 23. 15 ± 1. 05) mmol·L~(-1),respectively;creatinine( CREA) were( 11. 69 ± 0. 51) μmol·L~(-1),( 10. 23 ± 0. 28) μmol·L~(-1),( 10. 95 ± 0. 36) μmol·L~(-1),respectively; urinary albumin excretion rate( URE) were( 8. 96 ± 0. 84) μmol·L~(-1),( 13. 52 ± 0. 96) μmol·L~(-1),( 10. 22 ± 0. 69) μmol·L~(-1),respectively; the relative expression level of 78 kD-glucose regulated protein( GRP78)in normal,model and test groups were 0. 23 ± 0. 06,0. 79 ± 0. 22,0. 53 ± 0. 16,respectively; the relative expression level of PERK protein were 0. 19 ± 0. 04,0. 81 ± 0. 16,0. 62 ± 0. 19,respectively. There were statistically significant differences between the test group and the model group( P < 0. 05). Conclusion Astragalus membranaceus can reduce the body mass of DN mice,improve the degree of renal tissue damage and renal function,which may be related to the down-regulation of PERK pathway protein expression.
引文
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[6]姜文豪,孙艳,彭睿,等.miR-451通过靶向Psmb8抑制糖尿病肾病小鼠肾小球系膜细胞炎症反应[J].中国病理生理杂志,2018,34(3):494-499.
[7]张红红,卜祥伟,张建萍,等.糖肾清2号方对糖尿病肾病小鼠AMP激活蛋白激酶信号通路的影响[J].中国中西医结合杂志,2018,38(4):466-471.
[8]贺美芳.SiRNA干扰内质网应激PERK/e IF2α通路在糖尿病肾病中的作用及机制研究[D].太原:山西医科大学,2016.
[2]SZABAT M,PAGE M M,PANZHINSKIY E,et al.Reduced insulin production relieves endoplasmic reticulum stress and Induces cell proliferation[J].Cell Metabol,2016,23(1):179-193.
[3]NAKKA V P,PRAKASHBABU P,VEMUGANTI R.Crosstalk between endoplasmic reticulum stress,oxidative stress,and autophagy:potential therapeutic targets for acute CNS injuries[J].Molecul Neurobiol,2016,53(1):532-544.
[4]KLEIN J,RAMIREZ-TORRES A,ERICSSON A,et al.Urinary peptidomics provides a noninvasive humanized readout of diabetic nephropathy inmice[J].Kidney Internat,2016,90(5):1045-1055.
[5]卜灿,邓荃文,宋少练,等.复方黄甘对糖尿病肾病小鼠氧化应激和Klotho mRNA及蛋白表达的影响[J].中国临床药理学杂志,2016,32(24):2277-2281.
[6]姜文豪,孙艳,彭睿,等.miR-451通过靶向Psmb8抑制糖尿病肾病小鼠肾小球系膜细胞炎症反应[J].中国病理生理杂志,2018,34(3):494-499.
[7]张红红,卜祥伟,张建萍,等.糖肾清2号方对糖尿病肾病小鼠AMP激活蛋白激酶信号通路的影响[J].中国中西医结合杂志,2018,38(4):466-471.
[8]贺美芳.SiRNA干扰内质网应激PERK/e IF2α通路在糖尿病肾病中的作用及机制研究[D].太原:山西医科大学,2016.