摘要
目的：研究肾小管和肿瘤坏死因子与肾间质纤维化的关系。方法：肾小管上皮细胞及肾成纤维细胞的体外培养，采用逆转录－多聚酶链反应（ＲＴ－ＰＣＲ）、免疫组化染色、放射免疫测定（ＲＩＡ）及双重免疫组化染色技术，并测定３Ｈ－ＴｄＲ掺入率，观察ＴＮＦα与肾成纤维细胞增殖的关系。结果：肾小管上皮细胞既有ＴＮＦα－ｍＲＮＡ的表达，又有ＴＮＦ－Ｒ１ｍＲＮＡ的表达，还有ＴＮＦα的蛋白合成及分泌。且肾小管损伤及再生过程中，ＴＮＦα表达增加。对体外培养的大鼠肾成纤维细胞加入ＴＮＦα，３Ｈ－ＴｄＲ掺入率较对照组明显增高、肾成纤维细胞有ＴＮＦ－Ｒ１ｍＲＮＡ的表达。结论：损伤及再生的肾小管上皮细胞较正常时合成和分泌更多的ＴＮＦα，ＴＮＦα通过与ＴＮＦ－Ｒ的作用促进肾成纤维细胞的增殖。
OBJECTIVE To define the involvement of tumor necrosis factor (TNF) in renal interstitial fibrosis. METHODOLOGY Renal fibroblasts and tubular cells were cultured and characterized. Cell proliferation was measured by 3H TdR uptake assay. The effect of TNFα on renal fibroblasts proliferation was evaluated after the addition of 5μg/L or 40μg/L of TNFα. The expression of TNFα and its receptor 1 mRNAs in cultured tubular cells were identified by RT PCR. Cellular form of TNFα and TNFα released from cultured tubular cells were detected by immunostaining and radioimmunological assay separately. To study the association of tubular TNFα expression with acute renal injury, renal histology and immunohistochemistry were examined after the establishment of rat acute nephrotoxic model by gentamycin. RESULTS Both TNFα and its receptor 1 mRNAs were found to be expressed by renal tubular cells. Accordingly, cell bound TNFα and TNFα released to cultured supernatant were exhibited.TNFα expression by tubular cells were also found in vivo and there was an increase of TNFα immunostaining during cells recoverying from acute nephrotoxic injury. It is interesting to note that TNFα receptor 1 was also expressed by renal fibroblast. The addition of TNFα to the medium caused a significant increase of fibroblast proliferation. CONCLUSIONS TNFα is synthesized by tubular cells. TNFα derived from tubule may mediate the interaction between tubular cell and fibroblast and thus contribute to the tubulointerstitial pathology.
出处
《肾脏病与透析肾移植杂志》
CAS
CSCD
1998年第4期316-319,共4页
Chinese Journal of Nephrology,Dialysis & Transplantation
基金
国家自然科学基金
