Analysis Of Risk Factors And Mechanism Research For Postoperative DVT In Patients With Brain Tumor Surgery

Objective: To analyze the risk factors of postoperative DVT formation in patients with brain tumor surgery,explore the mechanism of influence of hypothermia during surgery on postoperative DVT formation,and the molecular mechanism of postoperative DVT formation induced by intraoperative hypothermia via KCNQ1OT1/mi R-146a-5p molecular axis.Methods:(1)Part I Case-control study: 231 cases of post-operative DVT were screened from brain tumor patients in a tertiary A general hospital in Xinjiang,and 231non-DVT patients were selected according to the principle of unmatched(grouped).The general clinical data,intraoperative situation,and laboratory test index were collected,and the risk factors of DVT formation were analyzed in patients with brain tumors after an operation.(2)Part II Clinical trial: Seventy-two patients with brain tumors were randomly selected and divided into a control group and an observation group.Routine and comprehensive heat preservation nursing measures were adopted respectively.The general clinical data,the incidence of postoperative DVT,blood flow velocity of lower limb veins,and the incidence of hypothermia before,during,and after the operation was compared in two groups.The expression levels of TNF-α,IL-6,IL-8,E-selectin,and PAI-1 in the serum of the two groups of patients were detected by ELISA,the expression of KCNQ1OT1 and PAI-1 in PBMC and mi R-146a-5p in peripheral blood serum were detected by q RT-PCR at three different 3-time points.(3)Part III Cell experiment:(1)The relationship was analyzed between mi R-146a-5p and PAI-1 targeting by 293 T through a double luciferase experiment.(2)HUVEC was cultured at 37℃,35℃,and 33℃ for 3,6,12,and 24 hours respectively.CCK-8 test was used to detect cell proliferation,the biochemical test was used to detect the activities of MDA and SOD in cells,and ELISA was used to detect TNF-α、 IL-6,IL-8,E-selectin,and PAI-1 levels in cell culture supernatant.Cultured at 37℃ and 35℃ for 6 and 12 hours respectively,the cell migration ability was detected by the Transwell test,and the expression level of KCNQ1OT1,PAI-1and mi R-146a-5p in cells was detected by q RT-PCR.(3)HUVEC plasmid transfection after 42 h was divided into 7 groups: blank group,hypothermic group,hypothermic+si RNA NC group,hypothermic+KCNQ1OT1-si RNA group,hypothermic+mi RNA inhibitor NC group,hypothermic+mi R-146a-5p inhibitor group,and hypothermic+KCNQ1OT1-si RNA+mi R-146a-5p inhibitor group.The blank group and other groups were cultured at 37℃ and 35℃ for 6 hours respectively.The cell proliferation was detected by CCK8,cell migration ability was detected Transwell chemotaxis test,and the formation of the cell lumen was measured by the Matrigel test.TNF-α,IL-6,IL-8,E-selectin,and PAI-1 productions in cell culture serum were detected by ELISA.p65 expressions in each group were detected by immunofluorescence assay.The expression levels of KCNQ1OT1,mi R-146a-5p,and PAI-1 in cells of each group were detected by q RT-PCR,and the expression of PAI-1,p65,p-p65,AKT,and p-AKT in cells of each group were detected by Western Blot.Results:(1)Part I:(1)The results of the univariate analysis showed that age,sex,operation duration,hypothermia during operation,preoperative core temperature,and APTT(P<0.05)had significant differences between DVT and non-DVT groups,were the risk factors of DVT formation in patients with a brain tumor after the operation.(2)Multi-factor analysis showed that over 45 years old,female,operation duration(>350min)and intraoperative hypothermia were risk factors(OR>1,P<0.05),APTT and preoperative core temperature were protective factors.(3)Taking age,operation duration,APTT,and intraoperative core temperature as test variables,the ROC curve was drawn.The AUC value of intraoperative core temperature was the largest,0.744,the maximum Jordan index was 0.368,the sensitivity was 60.6%,and the specificity was 76.2%,indicating a high diagnostic significance.(2)Part II:(1)The incidence of DVT in the control group was significantly higher than that in the observation group(P<0.05).(2)The blood flow velocity of patients in the two groups decreased significantly during and after the operation compared with that before the operation(P<0.05).The peak systolic flow rate and the peak diastolic flow rate in the observation group were significantly higher than those in the control group.(3)The incidence of hypothermia during and after the operation in the two groups was significantly higher than that before the operation(P>0.05).The incidence of hypothermia during and after the operation in the observation group was significantly lower than that in the control group(P>0.05).(4)The expression levels of TNF-α,IL-6,IL-8,E-Selectin,and PAI-1 in the serum of patients in the control group were significantly increased during and after the operation(P > 0.05).The postoperative concentration of IL-8 in the control group was significantly higher than that in the observation group(P > 0.05).(5)The postoperative expression levels of KCNQ1OT1 and PAI-1 in PBMC of patients were significantly higher after the operation in the two groups,and the expression level of both in the observation group was significantly lower than that in the control group(P<0.05).The postoperative expression level of mi R-146a-5p in the serum of patients in the two groups was significantly lower(P<0.05),and the expression level in the observation group was significantly higher than that in the control group(P>0.05).(3)Part III: 1)A negative targeting relationship was between mi R-146a-5p and PAI-1.2)(1)At the same time,the OD value of the 33℃ group was the lowest(P<0.05).At the same temperature,the longer the time,the higher the OD value would be.(2)With the decrease in temperature,the MDA activity increased significantly(P<0.05).At the same temperature,the longer the time,the higher the MDA activity would be.(3)The intracellular SOD activity in 33 ℃ and 35 ℃ groups was significantly lower than that in 37℃ group(P<0.05).(4)The concentrations of IL-6,IL-8,TNF-a,E-Selectin and PAI-1 in the cell supernatant increased significantly(P<0.05).(5)With the decrease in temperature,the cell migration ability decreased gradually(P<0.05),and treatment time had no significant effect on cell migration(P>0.05).(6)The expression of KCNQ1OT1 and PAI-1 in 35℃ group was significantly higher than that in37℃ group,and the expression of mi R-146a-5p was significantly lower(P<0.05).At the same treatment temperature,the expression level of the above molecules in different treatment time groups had no significant change(P>0.05).3)(1)The results of cell proliferation,migration,and lumen length showed that compared with the hypothermia group,the hypothermia+KCNQ1OT1-si RNA group significantly increased,hypothermia+mi R-146a-5p inhibitor group significantly decreased,hypothermia+KCNQ1OT1-si RNA+mi R-146a-5p inhibitor group was between the two groups(P<0.05).The indexes in the above groups were significantly lower than those in the blank group(P<0.05).(2)The intranuclear expression of p65,IL-6,IL-8,TNF-α,and E-Selectin concentration in cell supernatant: compared with the blank group,the concentrations of other groups were significantly higher except the hypothermia+KCNQ1OT1-si RNA group.Compared with the hypothermia group,the concentration of KCNQ1OT1-si RNA in the hypothermia+mi R-146a-5p inhibitor group decreased,while that in the hypothermia+mi R-146a-5p inhibitor group increased(P<0.05);The expression level of hypothermia+KCNQ1OT1-si RNA+mi R-146a-5p inhibitor group was between the two groups(P<0.05).(3)The expression of PAI-1and KCNQ1OT1 in the hypothermia+KCNQ1OT1-si RNA group was lower than that in the hypothermia group,The expression of mi R-146a-5p was up-regulated(P<0.001).The hypothermia+mi R-146a-5p inhibitor group was the opposite.The expression of PAI-1,KCNQ1OT1,and mi R-146a-5p in low temperature+KCNQ1OT1-si RNA+mi R-146a-5p inhibitor group was between the two groups(P<0.001).(4)Compared with the blank group,the expression of PAI-1 and p-p65 in the hypothermia group was significantly increased,and the expression of p-AKT was significantly decreased(P < 0.001);The expression of PAI-1 and p-p65 in hypothermia+KCNQ1OT1-si RNA group was significantly lower than that in hypothermia group,and the expression of p-AKT was significantly higher(P<0.05).(5)The expression of PAI-1 and p-p65 in the hypothermia+mi R-146a-5p inhibitor group was significantly higher than that in the hypothermia group,and the expression of p-AKT was significantly lower(P<0.05).The expression of PAI-1 and p-p65 in the hypothermia+KCNQ1OT1-si RNA+mi R-146a-5p inhibitor group was significantly higher than that in the hypothermia+KCNQ1OT1-si RNA group and lower than that in hypothermia+mi R-146a-5p inhibitor group(P < 0.05);The expression of p-AKT in this group was significantly lower than that in hypothermia+KCNQ1OT1-si RNA group and higher than that in hypothermia+mi R-146a-5p inhibitor group(P<0.05);There was no significant difference between p65 and AKT in each group(P>0.05).Conclusion:(1)Intraoperative hypothermia is closely related to postoperative DVT formation in patients with brain tumors.(2)Comprehensive thermal insulation nursing measures for patients undergoing brain tumor surgery can reduce the incidence of hypothermia during operation and prevent the formation of DVT after the operation.(3)Hypothermia affects the coagulation function by up-regulating the expression of KCNQ1OT1,inhibiting the expression of mi R-146a-5p,promoting the expression of PAI-1,activating NF-кB p65 signal pathway,inducing the expression of inflammatory factors and damage HUVEC function.This study reveals the molecular mechanism of DVT formation induced by hypothermia during operation in patients with brain tumors and provides a scientific basis for fully ensuring the safety of patients undergoing surgery and promoting the quality of their recovery from the perspective of nursing.