miR-92a和miR-362-3p调节TOB2参与亚低温对重型创伤性脑损伤保护作用

MiR-92a and miR-362-3p involved in the protective effects of mild hypothermia on severe traumatic brain injury via TOB2

目的探讨miR-92a和miR-362-3p调节TOB2参与亚低温对重型创伤性脑损伤保护作用。方法前瞻性选择2015年3月至2017年4月就诊的重型颅脑损伤患者84例,采用随机数字发将其分为常规治疗组(42例)和亚低温+常规治疗组(42例)。常规治疗组患者给予脱水、止血、抗炎及神经营养药物等常规治疗;亚低温+常规治疗组患者常规治疗基础上外加亚低温治疗。于治疗前,治疗后第1天、第7天和第14天采集肘静脉血5 ml,用qRT-PCR法检测患者血清miR-92a和miR-362-3p表达水平,用western blotting法检测患者血清TOB2蛋白表达,用荧光素酶活性实验验证TOB2是否是miR-92a和miR-362-3p的靶基因,记录患者伤后6个月时的格拉斯哥预后分级(GOS)。结果在治疗前,两组患者血清miR-92a和miR-362-3p表达无统计学差异(P>0.05)。在治疗后第1天、第7天和第14天,相比于常规治疗组,亚低温+常规治疗组患者血清miR-92a和miR-362-3p表达水平均降低(P<0.01),此外,治疗期间TOB2表达水平逐渐增加。荧光素酶活性实验验证证实ARNTL是miR-92a和miR-362-3p靶基因。在伤后第6个月,相比于常规治疗组,亚低温+规治疗组患者预后良好率和中等残疾率均增加,而重度残疾率、植物生存率和死亡率均降低(P<0.05)。结论创伤性脑损伤后miR-92a和miR-362-3p调节TOB2参与亚低温对其保护作用。

Objective To investigate whether miR-92 a and miR-362-3 p is involved in the protective effects of mild hypothermia on severe traumatic brain injury via TOB2.Methods 84 patients with severe traumatic brain injury(STBI) were selected in this study from March2015 to April 2017 and randomly divided into conventional treatment group(42 cases) and mild hypothermia + conventional treatment group(42 cases).The patients from conventional treatment group were given conventional treatment of dehydration,hemostatic and anti-inflammatory and neuro-trophic drugs,the patients from mild hypothermia + conventional treatment group were given conventional treatment,based on hypothermia treatment.5 ml of elbow vein bloods were collected before treatment,at first,seventh and fourteenth days after treatment,the serum levels of miR-92 a and miR-362-3 p were detected by qRT-PCR,the TOB2 protein was detected by western blotting,the luciferase reporter assay was used to verify TOB2 is target gene of miR-92 a and miR-362-3 p,the Glasgow outcome classification(GOS) was recorded at 6 months after injury.Results Before treatment,there was no significant difference in serum miR-92 a and miR-362-3 p between the two groups(P > 0.05).At the first,seventh and fourteenth days after treatment,compared with the conventional treatment group,the levels of serum miR-92 a and miR-362-3 p were decreased in mild hypothermia + conventional treatment group(P < 0.05),in addition,during the treatment,the TOB2 protein was gradually increased.The luciferase reporter assay confirmed that TOB2 was the target gene of miR-92 a and miR-362-3 p.At sixth months after injury,compared with the conventional treatment group,mild hypothermia + conventional treatment group had good prognosis and moderate residual rate,while severe disability rate,plant survival rate and mortality rate decreased(P < 0.05).Conclusion After traumatic brain injury,miR-92 a and miR-362-3 p is involved in the protective effects of mild hypothermia on severe traumatic brain injury via TOB2.