摘要
目的：研究碱性成纤维细胞生长因子（ｂＦＧＦ）在肾小管上皮细胞损伤和再生过程中的表达及其对肾小管上皮细胞的促增殖作用。方法：给大鼠腹腔注射庆大霉素４００ｍｇ（ｋｇ·ｄ），注射４天，造成肾小管损伤，分别于１、３、５、７、９、１２、１５天检查肌酐、尿素氮、乳酸脱氢酶及碱性磷酸酶含量，然后对肾损伤进行形态观察，用原位杂交和双重免疫组化染色观察ｂＦＧＦ表达与增殖的伴随情况，增殖细胞用增殖细胞核抗原（ＰＣＮＡ）显示。体外培养的人肾小管上皮细胞加入不同浓度的ｂＦＧＦ，测定３ＨＴｄＲ掺入率，观察ｂＦＧＦ与肾小管上皮细胞增生的关系。结果：生化检测尿ＬＤＨ、ＡＬＰ及血Ｃｒ、ＢＵＮ比对照显著增高。原位杂交及双重免疫组化染色显示：正常情况下，ｂＦＧＦ只在单核细胞、肾间质纤维细胞表达。肾小管损伤再生过程中，肾近曲小管上皮细胞表达增加，再生肾小管上皮细胞成熟后，不再表达ｂＦＧＦ。体外给培养的人肾小管上皮细胞加入ｂＦＧＦ后，３ＨＴｄＲ掺入率较对照组明显增高。结论：正常肾小管上皮细胞无ｂＦＧＦ产生，损伤及再生的肾小管上皮细胞有ｂＦＧＦ的基因表达和蛋白合成。ｂＦＧＦ可促进肾小管上皮细胞增殖。
OBJECTIVE Fibroblast growth factor (FGF) is a mitogenic cytokines for mesodermal and neuroectodermal cells, especially for fibroblasts. In recent years, the role of bFGF stimulating epithelial tissue proliferation was found. Cytokines participated in the injury and regeneration of tubular epithelial cells(TEC) often related with tubulointerstitial fibrosis. In this study, the action of bFGF in renal tubular injury and regeneration were investigated. METHODOLOGY Acute tubular necrosis was induced in rats by intraperitoneal injection of Gentamicin, 400 mg/(kg·d)×4d. Human TECs were separated and cultured. The levels of serum creatinine, BUN, Urine ALP and LDH in rats were measured. The morphologic changes in various phases were observed by light microscopy. To show the expression of bFGF gene and protein syntheris of bFGF in TECs, in situ hybridization and double immunohistochemic methods were used. The TEC proliferation was detected using incorporation of 3H TdR uptake method. RESULTS The concentrations of serum creatinine, BUN, urine ALP and LDH in experimental group were significantly higher than in controls. Focal tubular necrosis occured during 1st to 3rd days, diffuse necrosis appeard on 5th day. The regeneration of TEC was observed in 7th to 9th days accompanied with PCNA and bFGF double positive. During 12th to 15 th days, necrosis and regeneration in TEC only occured in regenerative phase (5th day). The uptake of 3H TdR was increased depentdent on addition of bFGF (0.5～5 μg/L) in cultured TECs. CONCLUSION bFGF is a mitogeneic cytokine to TEC. The expression of bFGF gene and the synthesis of bFGF occurred only in damaged and regenerative TEC. The bFGF not only stimulated gowth of TEC , but also possessed some significance for renal interstitial fibrosis.
出处
《肾脏病与透析肾移植杂志》
CAS
CSCD
1997年第3期221-224,共4页
Chinese Journal of Nephrology,Dialysis & Transplantation
基金
国家自然科学基金
