丹酚酸A对ZDF大鼠肾脏的保护作用和抑制脂蛋白相关磷脂酶A2的研究
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摘要
目的观察丹酚酸A(salvianolic acid A,SAA)对Zucker糖尿病肥胖(zucker diabetic fatty rat,ZDF)大鼠肾病的保护作用和对脂蛋白相关磷脂酶A2(lipoprotein-associated phospholipase A2,Lp-PLA2)表达的影响。方法取11～12周的雄性ZDF大鼠40只,根据体重与血糖,随机分为模型对照组、SAA 0. 25 mg/kg、0. 5mg/kg、1 mg/kg剂量组和阳性(25 mg/kg darapladib)对照组,每组8只,另取8只同周龄ZL大鼠作为正常对照组,各给药组每日给药一次,模型对照组和正常对照组每日给予同体积生理盐水,连续12周后,测定ZDF大鼠肾微循环血流速度,并检测血清中Lp-PLA2活性、尿素氮(blood urea nitrogen,BUN)、肌酐(creatinine,Crea),尿中转铁蛋白(urinary transferrin,TRF)、微量白蛋白(microalbumin,mALB)和视黄醇结合蛋白(retinol binding protein,RBP)的含量以及肾组织中Lp-PLA2 mRNA与蛋白表达,同时进行肾脏组织病理学观察。结果与正常对照组比,模型对照组大鼠出现明显的肾脏病理结构改变和肾微循环血流速度明显降低(P<0. 01),而血清Lp-PLA2活性、BUN水平以及尿中TRF、mALB和RBP均明显升高(P<0. 01),并且肾组织中Lp-PLA2 mRNA和蛋白表达升高(P<0. 01);给予SAA和Lp-PLA2抑制剂(darapladib)后能明显改善上述指标的改变,且SAA给药组中以0. 5mg/kg、1 mg/kg剂量组改善尤为显著。结论 SAA对ZDF大鼠肾脏有明显的保护作用,其机制可能与抑制LpPLA2有关。
Objective To observe the protective effect of salvianolic acid A( SAA) on nephropathy and the expression of lipoprotein-associated phospholipase A2( Lp-PLA2) in Zucker diabetes fatty rats( ZDF). Methods According to the body weight and blood glucose,40 male ZDF rats aged 11-12 weeks were randomly divided into fivegroups( n = 8 per group),namely,a model group,0. 25,0. 5,and 1 mg/kg SAA groups,and a positive( Lp-PLA2 inhibitor at 25 mg/kg) group. Meanwhile,another eight ZL rats of the same age were used as a normal group. Each drug administration group was given the corresponding drug once a day. Simultaneously,the model group and the normal group were administered normal saline of the same volume daily. After 12 weeks of continuous intervention, the renal microcirculation blood flow of ZDF rats was measured. Lp-PLA2 activity,blood urea nitrogen( BUN),and creatinine( Crea) in serum as well as transferrin( TRF),microalbumin( m ALB),and retinol binding protein( RBP) in urine were analyzed. Lp-PLA2 mRNA and protein expression in renal tissue was also examined. Moreover,renal histopathological observation was performed. Results Compared with the control group,the pathological structure of the kidney was clearly changed and renal microcirculation blood flow was significantly decreased in the model group( P< 0. 01),whereas serum Lp-PLA2 activity and BUN levels and urinary TRF,m ALB,and RBP were all notably increased( P<0. 01). Moreover,the expression of Lp-PLA2 mRNA and protein in renal tissues was markedly increased( P < 0. 01). The changes of these indicators were dramatically ameliorated after the administration of SAA and Lp-PLA2 inhibitor,especially in the groups with SAA at doses of 0. 5 and 1 mg/kg. Conclusions SAA can improve the symptoms of DN in type 2 diabetic rats,which may be related to the inhibition of Lp-PLA2 activity and mass.
Objective To observe the protective effect of salvianolic acid A( SAA) on nephropathy and the expression of lipoprotein-associated phospholipase A2( Lp-PLA2) in Zucker diabetes fatty rats( ZDF). Methods According to the body weight and blood glucose,40 male ZDF rats aged 11-12 weeks were randomly divided into fivegroups( n = 8 per group),namely,a model group,0. 25,0. 5,and 1 mg/kg SAA groups,and a positive( Lp-PLA2 inhibitor at 25 mg/kg) group. Meanwhile,another eight ZL rats of the same age were used as a normal group. Each drug administration group was given the corresponding drug once a day. Simultaneously,the model group and the normal group were administered normal saline of the same volume daily. After 12 weeks of continuous intervention, the renal microcirculation blood flow of ZDF rats was measured. Lp-PLA2 activity,blood urea nitrogen( BUN),and creatinine( Crea) in serum as well as transferrin( TRF),microalbumin( m ALB),and retinol binding protein( RBP) in urine were analyzed. Lp-PLA2 mRNA and protein expression in renal tissue was also examined. Moreover,renal histopathological observation was performed. Results Compared with the control group,the pathological structure of the kidney was clearly changed and renal microcirculation blood flow was significantly decreased in the model group( P< 0. 01),whereas serum Lp-PLA2 activity and BUN levels and urinary TRF,m ALB,and RBP were all notably increased( P<0. 01). Moreover,the expression of Lp-PLA2 mRNA and protein in renal tissues was markedly increased( P < 0. 01). The changes of these indicators were dramatically ameliorated after the administration of SAA and Lp-PLA2 inhibitor,especially in the groups with SAA at doses of 0. 5 and 1 mg/kg. Conclusions SAA can improve the symptoms of DN in type 2 diabetic rats,which may be related to the inhibition of Lp-PLA2 activity and mass.
引文
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[10]陆梦丽,吕礼应.脂蛋白相关磷脂酶A2检测方法及其临床应用进展[J].国际检验医学杂志,2018(18):2313-2316.
[11] Maeda H,Sogawa K,Sakaguchi K,et al. Urinary albumin and transferrin as early diagnostic markers of chronic kidney disease[J]. J Vet Med Sci,2015,77(8):937-943.
[12] Teng F,Yin Y,Cui Y,et al. Salvianolic acid A inhibits endothelial dysfunction and vascular remodeling in spontaneously hypertensive rats[J]. Life sci,2016,144(8):86-93.
[13]马全鑫,陈小真,戎亦骊,等.丹酚酸A改善ZDF大鼠视网膜病变及其作用机制[J].中国比较医学杂志,2017,27(10):40-46.
[14]陈小真,戎亦骊,马全鑫,等.糖尿病ZDF大鼠早期微血管并发症的生化指标及病理组织学变化[J].中国比较医学杂志,2016,26(05):63-70.
[15] Wang X,DuBois DC,Sukumaran S,et al. Variability in Zucker diabetic fatty rats:differences in disease progression in hyperglycemic and normoglycemic animals[J]. Diabetes Metab Syndr Obes.,2014,7:531-541.
[16]白静,刘易新,王旭东.糖化白蛋白及相关代谢指标与2型糖尿病早期肾功能损害发生的相关性研究[J].中国新药杂志,2018,27(16):1872-1876.
[17]芮旭辉.检测糖尿病肾病患者的肾功能指标在诊断其病情中的临床价值分析[J].当代医药论丛,2018,16(12):47-48.
[18] Williams ME. Diabetic nephropathy:the proteinuria hypothesis[J]. Am J Nephrol.,2005,25(2):77-94.
[19] Acharya NK,Qi X,Goldwaser EL,et al. Retinal pathology is associated with increased blood-retina barrier permeability in a diabetic and hypercholesterolaemic pig model:Beneficial effects of the LpPLA2 inhibitor Darapladib[J]. Diab Vasc Dis Res,2017,14(3):200-213.
[20] Wang Y,Li SS,Na SP,et al. Characterization of lipoproteinassociated phospholipase A2 in serum in patients with stage 3-5chronic kidney disease[J]. Am J Med Sci,2016,352(4):348-353.
[21] Canning P,Kenny BA,Prise V,et al. Lipoprotein-associated phospholipase A2(Lp-PLA2)as a therapeutic target to prevent retinal vasopermeability during diabetes[J]. Proc Natl Acad Sci USA,2016,113(26):7213-7218.
[22]苗艳,阎磊,朱清,等.血清脂蛋白相关磷脂酶A2水平对糖尿病肾病患者尿白蛋白排泄率和血肌酐的影响[J].新乡医学院学报,2015,32(06):522-525.
[23] Wang GH,Jin J,Sun LZ. Effect of lipoprotein-associated phospholipase A2 inhibitor on insulin resistance in streptozotocininduced diabetic pregnant rats[J]. Endocr J,2018,65(9):903-913.
[24] Fan HY,Yang MY,Qi D,et al. Salvianolic acid A as a multifunctional agent ameliorates doxorubicin-induced nephropathy in rats[J]. Sci Rep,2015,5:12273.
[2] Donate-Correa J,Martín-Nú1ez E,Muros-de-Fuentes M,et al.Inflammatory cytokines in diabetic nephropathy[J]. J Diabetes Res,2015,23(2):88-95.
[3]张作凯,王学凯,高振芳,等.丹酚酸A通过抗炎抗氧化发挥对急性肾损伤大鼠的保护作用[J].烟台大学学报(自然科学与工程版),2017,30(02):124-130.
[4] Zhang H,Liu Y,Jiang Q,et al. Salvianolic acid a protects RPE cells against oxidative stress through activation of Nrf2/HO-1signaling[J]. Free Radic Biol Med,2014,69(9):219-228.
[5]蔡红蝶.丹参茎叶对慢性肾功能损伤的改善作用及机制研究[D].南京中医药大学,2017.
[6] Wu P,Yan Y,Ma L,et al. Effects of the Nrf2 modulator salvianolic acid A alone or combined with metformin on diabetesassociated macrovascular and renal injury[J]. J BiolChem,2016,115(2):703-712.
[7]吴平,阎雨,侯碧玉,等.丹酚酸A增强二甲双胍对糖尿病血管病及肾病的保护作用[J].中国药理学通报,2015(b11):180-181.
[8] Hou B,Qiang G,Zhao Y,et al. Salvianolic acid a protects against diabetic nephropathy through ameliorating glomerular endothelial dysfunction via inhibiting AGE-RAGE signaling[J].Cell Physiol Biochem,2017,44(6):2378-2394.
[9]杨涛涛.丹酚酸A对STZ-DN大鼠的治疗作用研究[C].2014浙江省医学会临床药学分会、浙江省中西医结合学会中药分会学术会议论文汇编. 2014:2.
[10]陆梦丽,吕礼应.脂蛋白相关磷脂酶A2检测方法及其临床应用进展[J].国际检验医学杂志,2018(18):2313-2316.
[11] Maeda H,Sogawa K,Sakaguchi K,et al. Urinary albumin and transferrin as early diagnostic markers of chronic kidney disease[J]. J Vet Med Sci,2015,77(8):937-943.
[12] Teng F,Yin Y,Cui Y,et al. Salvianolic acid A inhibits endothelial dysfunction and vascular remodeling in spontaneously hypertensive rats[J]. Life sci,2016,144(8):86-93.
[13]马全鑫,陈小真,戎亦骊,等.丹酚酸A改善ZDF大鼠视网膜病变及其作用机制[J].中国比较医学杂志,2017,27(10):40-46.
[14]陈小真,戎亦骊,马全鑫,等.糖尿病ZDF大鼠早期微血管并发症的生化指标及病理组织学变化[J].中国比较医学杂志,2016,26(05):63-70.
[15] Wang X,DuBois DC,Sukumaran S,et al. Variability in Zucker diabetic fatty rats:differences in disease progression in hyperglycemic and normoglycemic animals[J]. Diabetes Metab Syndr Obes.,2014,7:531-541.
[16]白静,刘易新,王旭东.糖化白蛋白及相关代谢指标与2型糖尿病早期肾功能损害发生的相关性研究[J].中国新药杂志,2018,27(16):1872-1876.
[17]芮旭辉.检测糖尿病肾病患者的肾功能指标在诊断其病情中的临床价值分析[J].当代医药论丛,2018,16(12):47-48.
[18] Williams ME. Diabetic nephropathy:the proteinuria hypothesis[J]. Am J Nephrol.,2005,25(2):77-94.
[19] Acharya NK,Qi X,Goldwaser EL,et al. Retinal pathology is associated with increased blood-retina barrier permeability in a diabetic and hypercholesterolaemic pig model:Beneficial effects of the LpPLA2 inhibitor Darapladib[J]. Diab Vasc Dis Res,2017,14(3):200-213.
[20] Wang Y,Li SS,Na SP,et al. Characterization of lipoproteinassociated phospholipase A2 in serum in patients with stage 3-5chronic kidney disease[J]. Am J Med Sci,2016,352(4):348-353.
[21] Canning P,Kenny BA,Prise V,et al. Lipoprotein-associated phospholipase A2(Lp-PLA2)as a therapeutic target to prevent retinal vasopermeability during diabetes[J]. Proc Natl Acad Sci USA,2016,113(26):7213-7218.
[22]苗艳,阎磊,朱清,等.血清脂蛋白相关磷脂酶A2水平对糖尿病肾病患者尿白蛋白排泄率和血肌酐的影响[J].新乡医学院学报,2015,32(06):522-525.
[23] Wang GH,Jin J,Sun LZ. Effect of lipoprotein-associated phospholipase A2 inhibitor on insulin resistance in streptozotocininduced diabetic pregnant rats[J]. Endocr J,2018,65(9):903-913.
[24] Fan HY,Yang MY,Qi D,et al. Salvianolic acid A as a multifunctional agent ameliorates doxorubicin-induced nephropathy in rats[J]. Sci Rep,2015,5:12273.