摘要
目的探讨舒洛地特对单侧输尿管梗阻(UUO)小鼠肾脏间质纤维化的作用,及其对肾小管上皮-间充质转化(EMT)和肾组织Jagged-1、Notch1表达的影响。方法雄性CD1小鼠60只随机分为:假手术组(A组),仅行腹膜切开后缝合;对照组(B组),行单侧输尿管结扎,术后予生理盐水0.1ml/d皮下注射;低剂量治疗组(C组),单侧输尿管结扎术后予小剂量舒洛地特6mg/(kg·d)皮下注射;高剂量治疗组(D组),单侧输尿管结扎术后予大剂量舒洛地特12mg/(kg·d)皮下注射。B、C、D组动物分别于术后第3d、7d、14d处死6只,取输尿管结扎侧肾组织。肾脏标本行HE、Masson染色检测肾间质纤维化程度。应用免疫组化、WesternBlot、RT-PCR检测肾脏组织中α-SMA、TGF-β1、VEGF、Jagged-1和Notch1表达。结果 HE染色显示,UUO术后肾脏间质区域细胞外基质成分逐渐增多,肾小管萎缩,细胞成分减少,而肾小球病变不明显。舒洛地特治疗后UUO小鼠肾脏组织学变化减轻,高剂量组效果更为显著。Masson染色显示,UUO术后小鼠肾组织出现明显肾间质纤维化,舒洛地特治疗可明显抑制肾间质纤维化(P<0.001)。与A组比较,B组肾组织α-SMA、TGF-β1和Jagged-1mRNA及蛋白表达明显升高(P<0.05);舒洛地特治疗明显抑制UUO小鼠肾组织α-SMA、TGF-β1和Jagged-1表达(P<0.05)。B组和A组比较,VEGF表达明显降低(P<0.05),舒洛地特治疗后VEGF表达无明显变化。未检测出小鼠肾组织中Notch1表达。结论舒洛地特可抑制UUO小鼠肾间质纤维化,这可能与舒洛地特抑制UUO小鼠肾组织EMT有关,而EMT的减轻可能与UUO小鼠肾组织TGF-β1,Jagged-1表达下调有关,VEGF在其中的作用尚不明确,其确切机制需进一步探讨。
Objective To investigate the effect of sulodexide on renal interstitial fibrosis and the expression of Jagged-1,Notch1,vascular endothelial growth factor （VEGF）in unilateral ureteral obstruction （UUO） mice,and try to reveal the possible underlying mechanisms. Methods Sixty mice were divided into four groups randomly：group A （sham） was subjected to surgical manipulation but without ureteral ligation; group B（UUO＋saline） was subjected to ureteral obstruction and received saline subcutaneously （s.c.） once daily; group C（UUO＋LS） was subjected to ureteral obstruction and received sulodexide（6mg/kg） s.c. once daily; group D（UUO＋HS） was subjected to ureteral obstruction and received sulodexide（12mg/kg） s.c. once daily. Mice were sacrificed at day 3,7 and 14 after operation respectively. The severity of renal interstitial fibrosis was determined by pathologic techniques. The content of -SMA,TGF-β1,VEGF,Jagged-1 and Notch1 were determined in the kidneys by immunohistochemical or biochemical techniques. Results The severity of interstitial fibrosis,expression of -SMA,TGF-β1 and Jagged-1 were increased,whereas expression of VEGF decreased significantly in obstructed kidney compared to sham-operated kidney. Treatment with sulodexide attenuated interstitial fibrosis,reduced expression of -SMA,TGF-β1 and Jagged-1 after UUO operation dramatically,without significant affect on the expression of VEGF. Expression of Notch1 was not detectable in CD1 mice kidneys by RT-PCR and Western Blot. Conclusions Sulodexide diminishes interstitial fibrosis in obstructed kidneys. Inhibition of EMT is most likely to be included,which may be related to downregulating the synthesis of TGF-β1 and Jagged-1. The mechanisms underlying these effects remain to be elucidated.
出处
《北京医学》
CAS
2011年第2期157-162,166,共7页
Beijing Medical Journal
