The Study Of Inflammation Related Signal In The Renal Ischemia Reperfusion Injury Mouse Model

Objective:To observe the change of the level of serum of TNF-a,IL-1, IL-6 and GM-CSF and the level of TLR4 m RNA, TNF-a m RNA, IL-1a m RNA、IL-6 m RNA and CSF2 m RNA in kidney after Ischemic-reperfusion injury, discuss the role of the TLR and inflammatory reaction signal in the pathogenesis of RIRI.Methods:Renal IRI models were made in C57BL/6 mice(block renal pedicles for 45 mins and then loosen to make kidney reperfuse). All mice were divided into five groups: control(N), sham-operation group(S), I/R 4h, I/R 12 h, I/R 24 h. shamoperation group with same operation with I/R 4h, I/R 12 h,I/R 24 h but no block renal pedicles. Serum TNF-a, IL-1, IL-6 and GM-CSF were measured by ELISA, and the level of TLR4 m RNA、TNF m RNA、IL-1a m RNA、IL-6 m RNA and CSF2 m RNA were observed by QRT- PCR.Result:1. The levels of serum TNF-a、IL-1、IL-6、GM-CSF in I/R 4h, I/R 12 h, I/R 24 h were both higher than those in control group, I/R 12 h increase markedly, IL-1、IL-6、GM-CSF level increase much markedly than TNF-a(P<0.05).2. The levels of TLR4 m RNA、IL-1a m RNA、IL-6 m RNA and CSF2 m RNA in kidney were higher than those in control group, increased significantly in I/R 12h(P<0.05). TNF m RNA show no significant change. There was no significant difference between control group and sham-operation group. TLR4 m RNA level in kindey showed positive correlations with IL-1 m RNA(r=0.793, P=0.002);TLR4 m RNA level was also positively correlated with IL-6 m RNA(r=0.759, P=0.004) and CSF2 m RNA(r=0.772, P=0.003). No significant correlations were observed between TLR4 m RNA level and TNF m RNA.Discussion:1. I/R 4h, I/R 12 h, I/R 24 h had higher TNF-a、IL-1、IL-6、GMCSF levels than control group. I/R 12 h had the highest levels,which show that TNFa、IL-1、IL-6、GM-CSF contribute to the kidney injury occurred in early of Renal ischemia reperfusion.2. The level of TLR4 m RNA same as IL-1a m RNA、IL-6 m RNA and CSF2 m RNA in kidney in I/R 4h, I/R 12 h, I/R24 h were higher than control group, highest in I/R 12 h group, down-regulate as the reperfusion time extended. The level of TLR4 m RNA after ischemic-reperfusion injury in kindey was also positively correlated with IL-1 m RNA, IL-6 m RNA and CSF2 m RNA, which proved that TLR4 may work together with related inflammatory signaling pathway, leading to the injury occurred in the process of Renal ischemia reperfusion.