解毒通络保肾法对实验性DM大鼠肾足细胞凋亡相关因子Bax、Bcl-2、Caspase-3的影响
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摘要
目的探讨解毒通络保肾法对实验性糖尿病(DM)大鼠肾足细胞保护作用机制。方法 60只Wistar大鼠,雌雄各半,给予高糖高脂饲料,适应性喂养1 w,随机分为阴性对照组(A组,n=10),其余为造模组。喂养4 w后,造模组用链脲佐菌素50 mg/kg腹腔注射造模;造模成功的大鼠随机分为模型组(B组,n=10)、中药组(C组,n=10)。C组给予解毒通络保肾散每次4 ml,日灌胃1次;A、B组大鼠给予等体积蒸馏水。灌胃8 w后,观察各组大鼠空腹血糖(FPG)、血清肌酐(Scr)、尿素氮(BUN)、尿微量白蛋白(mAlb)的变化情况,取肾脏,制备肾组织匀浆。采用酶联免疫吸附试验检测大鼠肾脏足细胞凋亡相关因子Bax、Bcl-2及半胱氨酸天冬氨酸特异性蛋白酶(Caspase)-3蛋白表达水平。结果与A组相比,B、C组Bax、Caspase-3及Bax/Bcl-2表达均显著增加,Bcl-2表达显著降低(P<0.05,P<0.01);与B组相比,C组Bax、Caspase-3及Bax/Bcl-2表达均显著降低,Bcl-2表达显著上升(P<0.01,P<0.05)。结论解毒通络保肾法能够降低实验性DM大鼠空腹血糖,保护肾功能,减少尿蛋白的产生,其机制可能是下调Bax表达,上调Bcl-2表达,致Bax/Bcl-2比例下降,从而减少细胞凋亡执行蛋白酶Caspase-3的产生,起到抑制足细胞凋亡的作用,保护肾小球滤过屏障。
引文
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11 王彤,刘存志,刘玉珍,等.bcl-2/bax基因调控机体细胞凋亡的机制研究进展[J].中国老年学杂志,2008;28(8):1658-60.
12 Fossati S,Cam J,Meyerson J,et al.Differential activation of mitochondrial apoptotic pathways by vasculotropic amyloid-beta variants in cells compos-ing the cerebral vessel walls[J].FASEB J,2010;24(1):229-41.
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14 何泽,南征.解毒通络保肾散对实验性糖尿病大鼠肾脏功能和结构的影响[J].中国实用医药,2009;4(20):159-60.
15 何泽,南征,李才.解毒通络保肾散对晚期糖基化终末产物肾毒性影响的实验研究[J].长春中医药大学学报,2008;25(4):480-2.
2 Khoshnoodi J,Sigmundsson K,Ofverstedt LG,et al.Nephrinpromotes cell-cell adhesion through homophilic interactions[J].Am J Pathol,2003;163(6):2337-46.
3 李黎莉,陈志强,王月华,等.糖尿病大鼠足突间隙宽度与尿nephrin的相关性及益气养阴活血通络方的影响[J].时珍国医国药,2014;25(1):38-40.
4 Siu B,Saha J,Smoyer WE,et al.Reduction in podocyte density as a pathologic feature in early diabetic nephropathy in rodents:prevention by lipoic acid treatment[J].BMC Nephrol,2006;7:6.
5 赵承雅,韩香莲,南征,等.榛花消肾安胶囊干预实验性糖尿病大鼠足细胞相关蛋白α-actinin-4的研究[J].天津中医药,2016;33(7):440-4.
6 Sachs N,Sonnenber GA.Cell-matrix adhesion of podocytes in physiology and disease[J].Nat Rev Nephrol,2013;9(4):200-10.
7 Goto H,Wakui H,Komatsuda A,et al.Renal alpha-actinin-4:purification and puromycin aminonucleoside-binding property [J].Nephron Exp Nephrol,2003;93(1):27-35.
8 Roselli S,Heidet L,Sich M,et al.Early glomerualr filtration defect and sever renal disease in podocin-deficient mice[J].Mol Cell Biol,2004;24(2):550-60.
9 Economou CG,Kitsiou PV,Tzina AK,et al.Enhanced podocalyxin expression alters the structure of podocyte basal surface[J].J cell Sei,2004;117(Pt15):3281-94.
10 朱海慧,叶学锋.清热益气通络方对糖尿病肾病大鼠肾小球 Podocalyxin、Desmin 表达的影响[J].西安交通大学学报(医学版),2014;41(2):362-5.
11 王彤,刘存志,刘玉珍,等.bcl-2/bax基因调控机体细胞凋亡的机制研究进展[J].中国老年学杂志,2008;28(8):1658-60.
12 Fossati S,Cam J,Meyerson J,et al.Differential activation of mitochondrial apoptotic pathways by vasculotropic amyloid-beta variants in cells compos-ing the cerebral vessel walls[J].FASEB J,2010;24(1):229-41.
13 Zehendner CM,Librizzi L,de Curtis M,et al.Caspase-3 contributes toZO-1 and Cl-5 tight-junction disruption in rapid anoxic neurovascular u-nit damage[J].PLoS One,2011;6(2):e16760.
14 何泽,南征.解毒通络保肾散对实验性糖尿病大鼠肾脏功能和结构的影响[J].中国实用医药,2009;4(20):159-60.
15 何泽,南征,李才.解毒通络保肾散对晚期糖基化终末产物肾毒性影响的实验研究[J].长春中医药大学学报,2008;25(4):480-2.