利用转录组测序生物信息学初步探讨热射病预后关键基因

Identification of key genes for the heat stroke prognosis according to transcriptome sequencing

目的分析热射病患者外周血单个核细胞转录组测序生物信息学结果,寻找影响热射病预后的关键基因。方法选取2019-06-01~2020-08-30空军军医大学附属西京医院急诊科收治的热射病患者,按照预后分为生存组和死亡组。随机数字法在两组中分别选取患者进行外周血单个核细胞转录组测序,生物信息学分析寻找差异基因;RT-PCR验证筛选出的差异基因在生存组和死亡组中的表达。结果29例诊断为热射病患者入选,按照预后分为生存组和死亡组,其中生存组18例,死亡组11例。随机数字法选取6例生存患者和4例死亡患者,外周血单个核细胞进行转录组测序,两组患者有693个基因表达有明显差异,通过生物信息学分析筛选出趋化因子(C-C基元)配体5(CCL5)、基质金属蛋白酶8(MMP8)、MMP9基因可能与预后相关。将两组外周血单个核细胞的目标基因表达情况通过RT-PCR法表达检测,与生存组比较,死亡组MMP8基因(P=0.8955)和MMP9基因(P=0.0597)表达无明显变化,CCL5基因表达明显上升(t=7.056,P<0.05)。结论CCL5基因可能是影响热射病预后的关键基因,需要进一步深入研究。

Objective To find key genes related to the heat stroke prognosis by transcriptome sequencing of peripheral blood mononuclear cells in the patients.Methods Heat stroke patients treated in emergency department of Xijing Hospital,Air Force Medical University from 2019-06-01 to 2020-08-30 were selected and divided into survival group and death group according to the prognosis.Peripheral blood mononuclear cells of patients were randomly selected to perform transcriptome sequencing.Bioinformatics analysis was applied to find differential genes;RT-PCR was performed to verify the expression of the differential genes screened in the two groups.Results A total of 29 patients were diagnosed with sepsis,which were divided into survival group(18 cases)and death group(11 cases)according to their prognosis.Six surviving cases and four dead cases were randomly selected to perform transcriptome sequencing,which showed that 693 genes were differentially expressed.By bioinformatics analysis,MMP8,MMP9 and chemokine(C-C motif)ligand 5(CCL5)were screened as target genes related to the prognosis;Compared with the survival group,the expression of MMP8(P=0.8955)and MMP9(P=0.0597)did not change significantly,and the expression of CCL5 gene was significantly increased in death group(t=7.056,P<0.05).Conclusions CCL5 may be a key gene affecting the prognosis of heat stroke,which need further research to verify.