不同严重程度脓毒症小鼠骨髓中性粒细胞PDL1的表达及其机制

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Abstract Objective: To evaluate programmed death ligand 1(PDL1) expression of bone marrow neutrophils during sepsis in mice with different severity and to explore possible mechanism. Methods: ① In vivo:Thirty C57/B6 mice(male) were randomly divided into sham operation group(Sham, n=10),mild cecal ligation perforation group(mild CLP,n=10) and severe cecal ligation perforation group(severe CLP,n=10).Liver and lung tissues were harvested 24 hours post operation to detect pathological changes. Meanwhile,the changes of PDL1 on mouse bone marrow neutrophils were detected in this model. With repeating above steps about groups and CLP model construction at 72 hours post operation,the survival rate analysis was performed.② In vitro: Mouse bone marrow neutrophils were isolated and then divided into control group,lipopolysaccharide(LPS,1μg/mL) stimulation group.The function of neutrophil chemotaxis and phagocytosis were detected.In addition,PDL1 on neutrophils was detected in control group, LPS stimulation group, SB 203580 (10 μmol/L) group and SB 203580 (10 μmol/L)+LPS group,respectively. Results: ① In vivo: The survival rate of the mild CLP model group(40%, P<0.05)and the severe survival rate (0%,P<0.01) were significantly lower than that in sham operation group（100%）.At the same time pathological score was 1.250±0.677 0 in liver of mild CLP group(P<0.01), was 1.600±0.843 3 in lung of mild CLP group(P<0.01),was 2.900±0.614 6 in the liver of severe CLP group (P<0.01) and was 3.000±0.666 7 in the lung of severe CLP group(P<0.01), compared with the sham operation group. With detecting the ratio of bone marrow neutrophils, the proportion of mild CLP group was (37.57±1.89)%,while that of severe CLP group was (10.4±0.877 5)% ,which were significantly lower than that of sham operation group \[(59.27±1.168)%, P<0.01\].With detecting the ratio of PDL1 on neutrophils, the proportion of mild CLP group was(2.130±0.37)% while that of severe CLP group was (4.587±0.501 2)%,which were significantly lower than that of sham operation group \[(0.733 3± 0.1097)% ,P<0.01\]. ② In vitro: In terms of chemotaxis function, compared with the control group\[(1 101.00±99.78)μm\], there was a significant decrease of the chemotaxis distance when neutrophils stimulated with LPS\[(660.6±120.8)μm,P<0.01\].In terms of phagocytic function, compared with the control group(656.7±60.54),there was a significant increase of the phagocytic function in mean fluorescence intensity when neutrophils stimulated with LPS(998.7±46.01,P<0.01).The positive expression rate of PDL1 on bone marrow neutrophils was (72.27±1.656)% with the stimulation of LPS for 12 hours,which was significantly higher than that in the control group\[(1.713±0.519 6)%,P<0.01\]. However,with using the inhibitor of P38 MAPK SB 203580,PDL1 positive neutrophils stimulated with LPS were declined\[(37.17±1.38)%, P<0.01\]. Conclusion: The infiltration of neutrophils into tissues and organs were enhanced with the severity of sepsis and PDL1 of neutrophils was highly expressed with the severity of sepsis in the same way.PDL1 on the surface of neutrophils during sepsis was regulated by the P38 MAPK pathway.

Received: 19 October 2018

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