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目的观察糖尿病肾组织肿瘤坏死因子(TNF-α)、核因子(NF-κB)的表达及其与肾组织细胞凋亡关系以及雷帕霉素对其的影响。方法将20只GK大鼠随机分为DM组(n=10)、DMR组(n=10),10只Wistar大鼠作为NC组。DMR组予以6mg/kg雷帕霉素灌胃,DM与NC组仅给予等量生理盐水灌胃。TUNEL法检测肾组织细胞凋亡。免疫组织化学法(IHC)检测肾组织TNF-α和NF-κB的表达。结果 TUNEL法显示,NC组肾组织未见明显的凋亡细胞;4周及8周DM组凋亡数呈进行性增多(P<0.01)。IHC显示,NC组肾组织TNF-α、NF-κ Bp65有轻微阳性表达;4周、8周DM组NF-κBp65、TNF-α表达有逐渐上升的趋势,均显著高于正常对照组(P<0.01)。NF-κ Bp65、TNF-α阳性表达呈正相关(P<0.01);肾组织NF-α和NF-κ Bp65表达与肾组织细胞凋亡之间均呈正相关(P<0.01);与DM组比较,DMR组4周、8周组肾组织TNF-α和NF-κ Bp65的过度表达均被显著抑制(P<0.01),肾组织细胞凋亡数显著减少(P<0.01)。结论 NF-κB及其诱导的TNF-α在糖尿病肾病损伤中发挥重要作用,雷帕霉素可能通过抑制NF-κB和TNF-α的表达减少减轻肾组织细胞凋亡。
Objective To observe the expression of tumor necrosis factor (TNF-α), nuclear factor (NF-κB) and its relationship with renal cell apoptosis in diabetic nephropathy and the effect of rapamycin on it. Methods 20 GK rats were randomly divided into DM group (n = 10), DMR group (n = 10) and 10 Wistar rats as NC group. DMR group was given 6mg / kg rapamycin gavage, DM and NC group were given only the same amount of saline gavage. TUNEL assay of renal cell apoptosis. Immunohistochemistry (IHC) was used to detect the expression of TNF-α and NF-κB in renal tissue. Results TUNEL method showed that there was no obvious apoptotic cells in NC group. The number of apoptotic cells in DM group increased progressively at 4 weeks and 8 weeks (P <0.01). IHC showed that there was slight positive expression of TNF-α and NF-κBp65 in NC group; the expression of NF-κBp65 and TNF-α in DM group increased gradually at 4 weeks and 8 weeks, which were significantly higher than those in normal control group <0.01). The positive expressions of NF-κBp65 and TNF-αwere positively correlated (P <0.01). There was a positive correlation between the expression of NF-|Á and NF-|ÊBp65 in renal tissue and the apoptosis of renal tissue (P <0.01) The overexpression of TNF-α and NF-κBp65 in renal tissue of DMR group was significantly inhibited at 4 and 8 weeks (P <0.01), while the number of apoptotic cells in renal tissue was significantly decreased (P <0.01). Conclusion NF-κB and its TNF-α play an important role in the pathogenesis of diabetic nephropathy. Rapamycin may reduce the apoptosis of renal tissue by decreasing the expression of NF-κB and TNF-α.