Blocking MyD88 To Explore The Long-Term Effects And Mechanism Of The TLR Signaling Pathway In Renal Ischemia Reperfusion Injury Of Mice

[Objective] Apply innovative My D88 inhibitor TJ-M2010-2 and My D88 KO mice to study TLR / My D88 signaling pathway for the long-term effects of mouse renal ischemia-reperfusion injury.[Methods] In vivo, useing male BALB / C mice as experimental subjects, divided into four groups: SHAM group, ischemia-reperfusion injury in IRI group, TJ-M2010-2 group, My D88 KO groups, and after reperfusion of 24 h, 3d, 7d, 14 d, 28 d, obtain peripheral blood and left kidney,six mice each group,respectively. TJ-M2010-2 group at the time of IR-3,-2,-1,0,1,3,5,7,14 day with 0.5% carboxymethyl cellulose(CMC) dissolved to 100 mg / kg intraperitoneally parallel IRI, row midline laparotomy at 32 ? temperature inside line on the left kidney of free association after nondestructive vascular clamp block 60 min open. The remaining group received an equal amount of CMC and similar operations. ELISA to detect the 24 h inflammatory cytokines IL-1?, IL-6, TNF-? and MCP-1 levels. HE staining pathological damage, MPO immunofluorescence reaction inflammatory cell infiltration. ELISA, Fibronectin, Collagen IV and ?-SMA immunohistochemistry, to detect the 28 d serum levels of TGF-?1 and fibrosis. Another 28 d in each group after 6 rows right kidney removed 24 h, ie 29 d group. Detection of creatinine and urea nitrogen in serum. In vitro, HK-2 cells, as experimental subjects, with TGF-?1 induced undergoes the way of epithelial-mesenchymal technology(EMT),and divide into control group, TGF-?1 group and TJ-M2010-2 plus TGF-?1 group, light microscopy cell morphology was observed, Western blot detection of E-cadherin, Vimentin and ?-SMA expression.[Results] TJ-M2010-2 group My D88 KO group contrast to IRI group significantly reduced, in the 24 h of IL-1?, IL-6, TNF-?, MCP-1 lever; tissue damage and inflammatory cell infiltration significantly reduced. Serum levels of TGF-?1 and fibrosis in the 28 d was significantly reduced. Serum creatinine and urea nitrogen in the IRI was no difference at 24 h, rather TJ-M2010-2 group and My D88 KO group more reduction than IRI group in 29 d. In vitro, TJ-M2010-2 plus TGF-?1 group contrast to the TGF-?1 group,the cells form is smaller qualitative level, with the performance more expression of E-cadherin protein, less expression of Vimentin and ?-SMA.[Conclusion] TJ-M2010-2 IRI can reduce postoperative inflammation and tissue damage, and may reduce the long-term fibrosis. In vitro TJ-M2010-2 is capable of inhibiting process that HK-2 cells undergoing EMT.