Toll样受体系统在肾脏衰老中的表达变化

摘要

Objective To observe the expression changes of Toll -like receptors in aging process of rat kidneys.Methods Western blot and qRT-PCR were used to analyzed the expression changes of TLRs, NF-ΚB signal pathway molecules and inflammatory factors including CCL3, CCL4, CCL5, CD80, TNF-α and IL-12b in the kidney tissues from rats in 3-month-old group (young group, n = 20), 12-month-old group (middle group, n = 20) and 24-month-old group (old group, n = 20).Results ①The weight, kidney weight, TG and urinary albumin/creatinine ratio in 12 -month-old group [(466.86±32.62) g, (43.25±0.50) g, (2.32±0.46) mmol/L, (164.81 ±38.31) mg/mmoL] and 24-month-old group [(617.8±57.34) g, (5.01±1.00) g, (3.22±0.82) mmol/L, (256.00±49.39) mg/mmoL] were all higher than those in 3-month-old group [(236.3±12.28) g, (1.67±0.68) g, (1.20±0.32) mmol/L, (146.01 ±22.72) mg/mmoL], the differences were statistically significant (P < 0.05); BUN in 24-month-old group [(6.86±1.10) mmol/L] was higher than that in 3-month-old group [(5.73±0.47) mmol/L], the difference was statistically significant (P < 0.05); the differences of SCR, GLU, CHOL in the 3 groups were not statistically significant (P > 0.05); kidney weight and urinary albumin/creatinine ratio in 24-month-old group [(5.01±1.00) g, (256±49.39) mg/mmoL] were higher than those in 12-month-old group [(3.25±0.50) g, (164.81 ±38.31) mg/mmoL], the differences were statistically significant (P < 0.05).②TLR2, TLR3, TLR4, TLR5, TLR11 in 12-month-old group and 24-monlh-old group were all higher than those in 3-month-old group, TLR1 and TLR3 in 24-month-old group were higher than those in 3-month-old group, TLR9 in 24 -month-old group were higher than those in 3-month-old group, the differences were statistically significant (P < 0.05); the differences of TLR6, TLR7, TLR10 in 3 groups were not statistically significant (P > 0.05); TLR8 was not detected; TLR1, TLR2, TLR3, TLR4, TLR5, TLR11 in 24-month-old group were higher than those in 12-month-old group, the differences were statistically significant (P < 0.05).③IK-Bα and Phospho-NF-κB p65 in 24-month-old group and 12-month-old group were higher than those in 3-month-old group, the differences were statistically significant (P < 0.05); NF-ΚB p65 in 24-month-old group was higher than that in 3-month-old group, the difference was statistically significant (P < 0.05); NF-ΚB p65 and Phospho-NF-κB p65 in 24-month-old group were higher than those in 12-month-old group, the difference was statistically significant (P < 0.05).④CCL3, CCL4, CCL5, CD80, TNF-α and IL-12b in 24-month-old group were higher than those in 3-month-old group, the differences were statistically significant (P < 0.05); CD80 and IL-12b in 12-month-old group were lower than those in 3-month-old group, the differences were statistically significant (P < 0.05); CCL3, CCL4, CCL5, CD80, TNF-α and IL-12b in 24-month-old group were higher than those in 12-month-old group, the differences were statistically significant (P < 0.05).Conclusion TLRs may play an important role in the regulation of the kidney aging by activating NF-ΚB signal pathway and promoting the expression of inflammatory factors.%目的 观察Toll样受体(TLRs)系统在肾脏衰老中的表达变化.方法 用免疫印迹和实时定量PCR(qRT-PCR)技术分析3月龄组(青年组,n = 20)、12月龄组(中年组,n = 20)和24月龄组(老年组,n = 20)大鼠肾脏中TLRs系统分子、核转录因子(NF-κB)信号通路分子以及炎症因子包括趋化因子配体3(CCL3)、CCL4、CCL5、CD80、肿瘤坏死因子-α(TNF-α)和白细胞介素-12b(IL-12b)的表达变化情况.结果 ①12月龄组和24月龄组体重[(466.86±32.62)、(617.8±57.34)g]、肾重[(43.25±0.50)、(5.01±1.00)g]、三酰甘油(TG)[(2.32±0.46)、(3.22±0.82)mmol/L]和尿蛋白/肌酐比值[(164.81±38.31)、(256.00±49.39)mg/mmoL]高于3月龄组[(236.3±12.28)g、(1.67±0.68)g、(1.20±0.32)mmol/L、(146.01±22.72)mg/mmoL],差异有统计学意义(P < 0.05);24月龄组大鼠血尿素氮(BUN)水平[(6.86±1.10)mmol/L]高于3月龄组[(5.73±0.47)mmol/L],差异有统计学意义(P < 0.05);而血肌酐(SCR)、血糖(GLU)和胆固醇(CHOL)水平三组差异无统计学意义(P > 0.05).24月龄组肾重[(5.01±1.00)g]和尿蛋白/肌酐比值[(256±49.39)mg/mmoL]高于12月龄组[(3.25±0.50)g、(164.81±38.31)mg/mmoL],差异有统计学意义(P < 0.05).②12月龄组和24月龄组TLR2、TLR3、TLR4、TLR5、TLR11表达水平均高于3月龄组,24月龄组TLR1和TLR3水平高于3月龄组,24月龄组TLR9水平高于3月龄组,差异均有统计学意义(P < 0.05);三组TLR6、TLR7、TLR10水平差异无统计学意义(P > 0.05);TLR8未检测到.24月龄组TLR1、TLR2、TLR3、TLR4、TLR5、TLR11水平高于12月龄组,差异均有统计学意义(P < 0.05).③12月龄组和24月龄组IK-Bα和活化的磷酸化NF-κB p65(Phospho-NF-κB p65)水平高于3月龄组,差异有统计学意义(P < 0.05);24月龄组NF-κB p65水平高于3月龄组,差异有统计学意义(P < 0.05).24月龄组NF-κB p65和Phospho-NF-κB p65水平高于12月龄组,差异有统计学意义(P < 0.05).④24月龄组CCL3、CCL4、CCL5、CD80、TNF-α和IL-12b水平高于3月龄组,差异有统计学意义(P < 0.05);12月龄组CD80和IL-12b水平低于3月龄组,差异有统计学意义(P < 0.05).24月龄组CCL3、CCL4、CCL5、CD80、TNF-α和IL-12b高于12月龄组,差异有统计学意义(P < 0.05).结论 TLRs系统可能通过激活NF-κB信号通路、促进炎症因子的表达而在肾脏衰老过程中发挥重要作用.