摘要
目的研究白细胞介素１０（ＩＬ－１０）对人近端肾小管上皮细胞（ＨＫ－２细胞）在促炎症因子肿瘤坏死因子α（ＴＮＦα）作用下表达细胞间黏附分子１（ＩＣＡＭ－ｌ）及其相关核转录途径的影响。方法 用ＨＫ－２细胞作靶细胞，用细胞酶联免疫吸附法（ＥＩＪＳＡ）和Ｎｏｒｔｈｅｒｎ杂交观察ＩＣＡＭ－１的蛋白和基因的表达，以电泳迁移率变动法测定转录因子核因子ｋＢ （ＮＦｋＢ）和激活蛋白 Ｉ（ＡＰ－１）的活性。结果ＴＮＦα呈剂量依赖地诱导ＨＫ－２细胞ＮＦｋＢ的活化及ＩＣＡＭ－１的基因和蛋白表达，这些作用可以被ＮＦｋＢ的抑制剂对甲苯磺－Ｌ－苯丙氨酸氯甲基甲酮（ＴＰＣＫ）所抑制，但ＴＮＦα对ＨＫ－２细胞的ＡＰ－１活性无影响。 ＩＬ－１０（１～ ２０ｎｇ／ｍｌ）可抑制 ＴＮＦα诱导的 ＨＫ－２细胞 ＩＣＡＭ－ｌ基因和蛋白表达及 ＮＦｋＢ的活化。结论ＴＮＦα诱导人肾小管上皮细胞ＨＫ－２的ＮＦｋＢ活化，进而促进ＩＣＡＭ－ｌ基因和蛋白表达，ＩＬ－１０可抑制ＴＮＦα诱导的上述炎症效应。
Objecive To explore the effect of interleukin-10(IL10) on tumor necrosis factor-α(TNFα)- induced expression of intercellular adhesion molecule 1 (ICAM-1 )and related transcription pathway in human renal tubular cells. Methods Human renal proximal tubular cell line HK-2 cell was used as target cell. ICAM-1 was measured by cell ELISA and Northern blot in protein and mBNA levels. Activities of transcriptional factors nuclear factor GB(NFkB) and activator protein 1 (AP-1) were determined by electrophoresis mobility shift assay. Results TNFα-induced ICAM-l expression in HK-2 cells was increased in both protein and mNRA levels(P < 0.01 ). NFkB inhibitor N-tosyl phenyalanine chlormethyl ketone (TPCK)could inhibit these effects of TNLFα. TNFα had no effect on AP-l activity with doses of 1 ng/ml to 100 ng/ml and incubation periods of one to four hours. IL-10 iuhibited TNFα-induced ICALM-1 protein expression in dosage from 1 ng/ml to 20 ng/ml(decreased by 25 .7% to 35. 1 %, P < 0.05), as well as ICAM-1 mRNA expression and activity of NFkB in HK-2 cells. Conclusions TNFα induces ICAM-l expression in HK-2 cells through activation of NFkB. Inhibitory effect of IL-10 on ICAM-l expression induced by TNFα is mediated by, at least in a part, down-regulation of NFkB activity.
出处
《中华肾脏病杂志》
CSCD
北大核心
2000年第1期16-19,共4页
Chinese Journal of Nephrology
基金
国家自然科学基金!(39570298)
关键词
肾小管间质病变
白细胞介素10
ICAM-1
HK-2细胞
Interleukin-10
Renal tubule
Epithelial cell
Interecllular adhesion molecule-l
Transcription factor
