Role Of TIL2 And TIL4 In Post-hemorrhagic Shock Mesenteric Lymph-mediated Inflammatory Response In Murine Kidneys

Background: Uncontrol inflammatory response is one of mechanisms by which severe hemorrhagic shock inducing acute kidney injury.Following hemorrhagic shock,the return of gut derived toxic substances contained mesenteric lymph to systemic circulation is a major pathway and mechanism of multiple organ injury.The interventions of post-hemorahgic shock mesenteric lymph(PHSML)return play positive effect of alleviating organ injury of hemorrhagic shocked animal.Objective: To reveal the role of toll-like receptor(TLR)-2 and TLR4 in PHSML-mediated inflammation in renal tissue,a murine model of hemorrhagic shock was employed in the present study,the effects of PHSML drainage on the expression of mRNA of TLR2 and TLR4 and the levels of interleukin(IL)-1? and IL-18 in renal tissue were observed,and the levels of IL-1? and IL-18 in renal tissue were detected after intravenous injection of PHSML to normal wild type(WT),TLR2-/-and TLR4-/-mice.Methods: Eighteen male C57BL/6J mice were randomized to sham,shock,and shock+drainage groups.The model of hemorrhagic shock was established in the shock and shock+drainage groups.As following: a hypotension of 40±2 mmHg was reduced and maintained at this level for 90 min,followed by fluid resuscitation with the shed blood and an equal volume of Ringer's solution,then,the observation was performed for 3 hours after the end of infusion.In the shock+drainage group,after resuscitation finished,the mesenteric lymphatic duct was cannulated and mesenteric lymph was drained up to 3 h with mesenteric lymph duct intubation.At 3 h after resuscitation or corresponding time in each group,kidneys from a fixed location were harvested for the detection of TLR2 and TLR4 mRNA expressions using qRT-PCR method and the levels of IL-1? and IL-18 using ELISA method.In addition,the normal mesenteric lymph(NML)was drained from 18 male normal mice;meanwhile,the PHSML was drained from 12 mice subjected to hemorrhagic shock and fluid resuscitation from 0 to 3 h.Then,the NML and PHSML were injected intravenously to normal mice wild type(WT),TLR2-/-and TLR4-/-mice within 30 min,n=6/group,respectively.After 2.5 h,the renal tissues were harvested for the determination of IL-1? and IL-18.Results: The mRNA expressions of TLR2 and TLR4 and the levels of IL-1? and IL-18 in renal tissue in the shock group were significantly increased compared to the sham group(P<0.05).PHSML drainage significantly decreased the mRNA expressions of TLR2 and TLR4 and the levels of IL-1? and IL-18 in renal tissue from the shocked mice(P<0.05).Intravenous injection of PHSML significantly increased the IL-1? and IL-18 levels in renal tissue of normal WT mice,increased the IL-18 level in renal tissue of TLR4-/-mice(P<0.05).However,PHSML treatment had no significant difference in the IL-1? and IL-18 levels in renal tissue of TLR2-/-mice,as well as the IL-1? level in renal tissue of TLR4-/-mice(P>0.05).Conclusion: TLR2 and TLR4 play important role in PHSML-mediated inflammation in renal tissue.