摘要
目的探讨帕立骨化醇对糖尿病肾病(DKD)肾小管上皮间充质转化(EMT)的作用及内在机制。方法将24只SD大鼠按随机数字表法分为帕立骨化醇干预组(P组)、糖尿病肾病组(D组)和正常对照组(C组),每组各8只。P组和D组大鼠腹腔注射链脲佐菌素65mg/kg建立糖尿病肾病模型,造模成功后第2天P组腹腔注射帕立骨化醇(溶于丙二醇中)0.4μg/kg,3次/周,D组予等体积丙二醇;C组仅予等体积丙二醇。4周后测血、尿生化指标;进行肾脏病理学检查;免疫组化及Western blot技术检测肾组织E-钙粘蛋白(E—cadherin)、。α-平滑肌肌动蛋白(α-SMA)、纤连蛋白(FN)、转化生长因子-β1(TGF—β1)及Klotho的表达,并进行指标间的相关性分析。结果 D组大鼠血清肌酐(Scr)、血尿素氮(BUN)及24 h尿蛋白水平均高于C组,P组均低于D组(均P<0.05)。C组大鼠肾小管结构完整清晰,无明显病理改变;D组可见肾小管间质局部炎症细胞浸润,肾小管上皮细胞脱落,小管扩张,基底膜断裂;P组肾小管间质病理改变较D组减轻。D组大鼠肾组织E—cadherin与Klotho表达低于C组,而P组高于D组(均P<0.05);与C组比较,D组大鼠肾组织α—SMA、FN及TGF—B 1表达增加,而P组表达均较D组减少(均P<0.05)。Klotho表达与E—cadherln呈正相关(r=0.924,P<0.05),而与α—SMA、FN及TGF—β1均呈负相关(r=-0.806、-0.623、-0.856,均P<0.05)。结论帕立骨化醇可抑制糖尿病肾病大鼠肾小管上皮EMT,其作用可能与增加肾组织Klotho表达,同时减少TGF—β1合成相关。
Objective To investigate the effect of parlicalcitol on tubular epithelial mesenchymal transition （EMT） and underlying mechanism in diabetic kidney disease （DKD）. Methods DKD was induced by single intraperitoneal injec- tion of 65 mg/kg streptozotocin（STZ） in 16 Sprague Dawley （SD） rats, then the animals were randomly divided into group P and group D with 8 in each. Rats in group P received intraperitoneal injection of paricalcitol（0.4 μg kg-1） in propylene glycol three times a week; rats in group D received intraperitoneal injection of same volume of propylene glycol; 8 rats in group C served as normal control and received propylene glycol as in group D. Blood, urine and renal tissue samples were collected after 4 weeks intervention of paricalcitol or vehicles. Biochemical indexes were measured, renal tissue was examined histopathologically and the expression of E-cadherin, alpha-smooth muscle actin （α-SMA）, fibronectin （FN）, transforming growth factor- β 1 （TGF- β1） and klotho in renal tissue were measured with immunohistochemistry and Western blotting, re- spectively. In addition, the correlation among the indexes was analyzed. Results Scr, BUN and 24 h urine protein in- creased significantly in group D compared with group C, while decreased in group P compared with group D （all P〈0.05）. Local inflammatory cell infiltration, tubular epithelial cell detachment, tubular expansion and basement membrane rupture were observed in group D, but attenuated in group P compared with group C. The expression of E-cadherin and klotho de- creased, while α-SMA, FN and TGF- β 1 increased in group D compared with group C （all P〈0.05）. Compared with D, the expression of E-cadherin and klotho increased, while α-SMA, FN and TGF-β 1 decreased in group P （all P〈0.05）. The expression of klotho was negatively correlated with α-SMA, FN and TGF-β 1, while was positively correlated with E-cad- herin （r=-0.806, P〈0.05; r=-0.623, P〈0.05; r=-0.856, P〈0.05; r=0.924, all P〈0.05）. Conclusion ParicaIcitol can inhibit tubular EMT in DKD, which may be associated with its effect of upregulating Klotho expression, and inhibiting TGF-β 1 syn- thesis.
出处
《浙江医学》
CAS
2015年第11期925-930,共6页
Zhejiang Medical Journal
基金
浙江省医药卫生科研基金(2015KYA201)
宁波市自然科学基金(2014A610245)
