Laws of macrophage polarization in rats with sepsis-induced acute kidney injury
Objective To observe the pathological changes of renal tissue and laws of polarization of renal macrophages after acute renal injury in rats with sepsis.Methods Rat sepsis models were constructed by cecal ligation and perforation.Rats in the sepsis groups were euthanized at 12,24,48,48 and 72 hours after surgery respectively,and then renal tissue samples were obtained.Rats in the sham-surgery group received the same treatment as the sepsis groups except cecal ligation and perforation,and then were euthanized at 24 hours after surgery for collecting renal tissue samples.The samples were stained by HE staining,and then the status of renal tissue edema was observed.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect mRNA expression in M1 and M2 macrophages in renal tissues.The protein expression of M1-type marker CD86 and M2-type marker CD206 was observed by Western blotting,and semiquantitative analysis was conducted by Image-Pro Plus software.Results Compared with the sham-surgery group,the volume of glomerulus cavity at 12,24,48 and 72 hours after surgery in the sepsis groups decreased,and the edema degree was significantly aggravated.The mRNA expression of M1-type surface marker CD 16(10.39±0.67),CD86(7.59±1.11)and iNOS(7.36±1.36)increased significantly at 12 hours after surgery,but gradually decreased over time(P＜0.001).The mRNA expression of M2-type surface marker Arg1(1.05±0.15),CD163(1.56±0.36)and IL-4(1.38±0.11)slightly increased at 12 hours after surgery,but the difference was not statistically significant(P＞0.05).The mRNA expression of M2-type surface marker Arg1,CD163 and IL-4 gradually increased at 24 and 48 hours after surgery,with statistically significant differences(P＜0.001).At 72 hours after surgery,the mRNA expression of Arg1(4.88±0.53),CD163(3.70±0.59)and IL-4(4.94±1.10)maintained the rise,showing statistically significant differences(P＜0.001).The detection results of Western blotting were consistent with those of qRT-PCR,and Western blotting displayed that the expression of Ml-type marker CD86(4.44±0.12)increased in the early postoperative period after sepsis(within 24 hours),but the expression of M2-type marker CD206(7.76±0.22)increased at 24-72 hours after surgery.Conclusion The laws of Ml and M2 macrophages polarization at different time points after sepsis-induced acute kidney injury provide a theoretical basis for the early intervention of renal injury after sepsis from the perspective of macrophage immune regulation.
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