| Background: In recent years,wasp stings are a serious threat to human life and health.The diversity of wasp venom determines that wasp stings are diseases that can affect the whole body,such as acute hemolysis,rhabdomyolysis,acute renal tubular necrosis,liver damage,myocardial damage,coagulation dysfunction,and central nervous system damage.It even leads to multiple organ dysfunction syndrome(MODS).Severe patients can go into a coma or even die.MODS is developed from systemic inflammatory response syndrome(SIRS).In the process of inflammatory response,inflammatory factors trigger immune response,resulting in microcirculation thrombosis and tissue damage.Interleukin-8(IL-8)is a CXC chemotactic cytokine with a variety of cellular sources.It can bind to IL8 receptors IL8RA(CXCR1)and IL8RB(CXCR2)on microvascular endothelial cells.In addition,different patients with wasp stings have varying degrees of disease development and prognosis,suggesting that may be related to individual genetic background.In this study,from the molecular expression and genetics,to explore the relationship between IL-8 and its receptor and SIRS in patients with wasp stings,and to analyze the risk factors of SIRS caused by wasp stings,so as to lay a foundation for further exploring the pathogenic mechanism of wasp stings.The purpose of this study is to provide a theoretical basis for clinicians to understand the development of the disease and early intervention measures.Objective: To investigate the expression level of IL-8 and its receptor in plasma of patients with wasp stings,to find out some susceptible genes of SIRS caused by wasp stings,and to study the relationship between gene polymorphism and SIRS caused by wasp stings.And the risk factors were analyzed.It lays a foundation for further enhance the understanding of disease development and provide theoretical basis for early clinical interventions.Methods: The patients with wasp stings were divided into the SIRS groupand the control group(the non-SIRS group).Collected the anticoagulant peripheral venous blood and separated the plasma and leukocytes.The whole genome DNA,was extracted from the leukocytes of the patients included in the study,and the IL-8(-251A/T,-738T/A,-845T/C,+396T/G),CXCR1(+860G/C),CXCR2(+811C/T,+1235T/C,+1440G/A)were selected for study.The target fragments were amplified by PCR(polymerase chain reaction)and sent the PCR amplified products to biological company for bidirectional DNA sequencing.All the results were compared with the corresponding base sequence fragments of human IL-8 and its receptor in Gene bank,all the SNPs on the sequencing map were found in the bank,and recording the genotyping.The plasma levels of IL-8,CXCR1 and CXCR2 were measured by enzyme linked immunosorbent assay(ELISA).Finally,the risk factors of wasp stings were analyzed by logistic linear regression.Results:(1)The plasma expression levels of IL-8,CXCR1 and CXCR2 in the SIRS group were significantly higher than in the control group(the non-SIRS group)(P<0.05).(2)No polymorphism was observed at-738T/A,-845T/C and +781C/T of IL-8 gene,but there were polymorphisms at-251A/T and +396T/G.In the control group(the non-SIRS group),the frequency variation of-251A/T and+396T/G genotypes did not deviate from the Hardy-Weinberg equilibrium((HWE)(P > 0.05).Three genotypes of AA,AT and TT were observed in-251A/T.There were significant differences in the frequencies of genotypes and alleles between the SIRS group and the control group(the non-SIRS group)(P<0.01).There were three genotypes of TT,TG and GG in +396T/G,but there was no significant difference in +396T/G polymorphism between the two groups(P>0.922).(3)There is no polymorphism was observed at the +860G/C and CXCR2+1440G/A in the CXCR1 gene,but there was polymorphism at the CXCR2+811C/T and +1235T/C.In the control group(the non-SIRS group),thevariation of +811C/T and 1235T/C genotype frequency did not deviate from the Hardy-Weinberg equilibrium(HWE)(P>0.05).Three genotypes of CC,CT and TT were observed in +811C/T and three genotypes of TT,TC and CC were observed in +1235T/C.There were significant differences in the frequencies of genotypes and alleles between the SIRS group and the control group(the non-SIRS group)(P<0.02;P<0.019).(4)Logistics regression analysis showed that IL-8-251 T allele(TT+AT),the number of stings ≥ 10,sex and limb stings were risk factors for SIRS(OR=3.971,[95%CI,1.618-9.743],P=0.003;OR=11.095,[95%CI,4.22-29.17],P<0.001;OR=2.418,[95%CI,1.081-5.408],P=0.032;OR=2.148,[95%CI,0.877-5.257],P=0.034)(5)Logistics regression analysis of CXCR2 +811T/C and related covariates showed that the number of sting≥10,sex,limb stings and CXCR2 +811T allele(TT+CT)were risk factors for SIRS(OR=10.606,[95%CI,3.993-28.168],P<0.001;OR=2.754,[95%CI,1.209-6.278],P=0.016;OR=2.656,[95%CI,1.061-6.649],P=0.037;OR=4.223,[95%CI,1.863-9.571],P=0.001).(6)Logistics regression analysis of CXCR2 +1235T/C and related covariates showed that the number of stings≥10,sex,limb stings and CXCR2+1235C allele(CC+TC)were risk factors for SIRS(OR=10.329,[95%CI,3.872-27.555],P<0.001;OR=2.768,[95%CI,1.218-6.294],P=0.015;OR=2.64,[95%CI,1.058-6.588],P=0.038;OR=4.012,[95%CI,1.773-9.079],P=0.001).Conclusion: We evaluated the association of genetic polymorphism of IL-8 and its receptors with SRIS caused by wasp stings.The expression level of plasma IL-8 and its receptor can be used as a predictor of SIRS in patients with wasp stings,IL-8-251 T,CXCR2 +811T,CXCR2 +1235C,the number of stings≥10,male patients and limb stings were risk factors for SIRS in wasp stings. |
