| Objective: Thist study intends to establish a rat model of hemorrhagic shock-reperfusion liver injury,to quantitatively evaluate the protective effect of estradiol on liver during resuscitation of hemorrhagic shock by virtual touch tissue imaging quantification(VTIQ)and contrast-enhanced ultrasound(CEUS),and to analyze the correlation between VTIQ and CEUS quantitative parameters and liver function index and enzyme-linked immunosorbent assay(ELISA)index.Finally,it intends to explore the application value of VTIQ and CEUS techniques in evaluating drug intervention on hemorrhagic shock-reperfusion liver injury.Methods: 30 healthy adult male SD rats were randomly divided into sham operation group(Sham group),hemorrhagic shock and reperfusion injury group(HS-RI group)and estradiol group(E group).In the HS-RI group,the left common carotid artery was separated from the deep side of the trachea in the muscular layer of the left neck,and 20 g indwelling needle was placed,and then the biosensor was connected with the three-way tube,and the initial mean arterial pressure(MAP)was recorded.Moreover,the left common femoral vein and common femoral artery were separated from the left inguinal area,and 24 g indwelling needles were placed respectively.After bleeding from the left common femoral artery to 40% of the initial MAP for 1 hour,the left common femoral vein was infused with resuscitation solution,and the blood pressure returned to 80% of the initial MAP,indicating that the HS-RI model was successfully constructed.Group E: the rats were subcutaneously injected with estradiol 1mg/kg after bloodletting to 40% of the initial MAP for 1hour,and the rest operation was the same as that of the HS-RI group.Sham group: rats were only intubated with left common carotid artery,common femoral artery and common femoral vein,without subsequent bloodletting and fluid replacement.VTIQ and CEUS examination were performed 12 hours after the establishment of the model.First of all,VTIQ examination was performed: with Siemens ACUSONS3000 ultrasound diagnostic instrument and 9L4 frequency conversion linear array probe,a clear two-dimensional image of the liver was obtained by intercostal scan in rats,and the VTIQ quality mode was selected.When the quality mode was the best,the velocity mode was adjusted to the velocity mode.The region of interest was placed in different positions of the liver at the same depth to measure the shear wave velocity(SWV).After VTIQ examination,the right common femoral vein was separated from the right inguinal area of rats and 24 G indwelling needle was placed to establish venography pathway.The instrument and probe used were the same as for the VTIQ examination,the built-in memory and timer were started while the contrast medium was injected into the right common femoral vein.The dynamic process of the successfully stored contrast was analyzed by ACQ quantitative analysis software,the time-intensity curve(TIC)was obtained,and four parameters were obtained: time to peak(Tp),mean transit time(MTT),area under curve(AUC),and peak intensity(Peak).The blood 3ml of inferior vena cava was taken by laparotomy after contrast-enhanced ultrasound in all three groups of SD rats.The levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in serum after centrifugation were detected by ADVIA2400 automatic biochemical analyzer.The rats were killed immediately after successful blood sampling,and two right liver tissues were quickly taken for hematoxylin-eosin staining and detection of inducible nitric oxide synthase(i NOS),B-lymphoma-2(Bcl-2),and interleukin-6(IL-6),respectively.The data were analyzed by SPSS25.0software package.The quantitative data with normal distribution were statistically described by "mean ± standard deviation".The indexes among the three groups were compared by single factor analysis of variance.The receiver working characteristic(ROC)curve of SWV for predicting acute liver injury in rats was drawn,the area under the curve was calculated and the cutoff value was obtained.Pearson linear correlation was adopted to analyze the correlation between VTIQ and CEUS parameters and ALT,AST,Bcl-2,IL-6,i NOS.P<0.05 indicated that the difference was statistically significant.Result:1.The results of VTIQ test: Compared with Sham group,SWVmin,SWVmax and SWVmean in HS-RI group and E group were increased(P<0.05).Compared with SWVmin(3.02±0.38)m/s,SWVmax(3.83±0.17)m/s,and SWVmean(3.51 ± 0.22)m/s in HS-RI group,SWVmin(2.65±0.13)m/s,SWVmax(3.34±0.25)m/s,and SWVmean(2.97±0.14)m/s in group E were significantly lower(P<0.05).ROC curve analysis showed that SWVmean had the largest area under the curve for predicting acute liver injury(0.990),with a cutoff value of 2.93m/s,a sensitivity of 100% and a specificity of 90%.2.Quantitative analysis of contrast-enhanced TIC curve: The contrast process was analyzed by ACQ software,and the TIC curve was obtained.The TIC curve of Sham group reached the peak faster than that of HS-RI group and E group,and then decreased rapidly.The TIC curve in group E increased and decreased faster than that of the HS-RI group.Compared with Sham group,Peak(58.87 ± 2.91)% and AUC(11937.27 ±567.37)%s in HS-RI group increased significantly,and Tp(49.94±3.36)s and MTT(152.53 ± 11.73)s in HS-RI group prolonged significantly(P<0.05).Compared with Sham group,Peak(41.11±2.93)% in E group decreased,AUC(5690.75±348.15)% s in E group increased,and Tp(36.42±3.80)s and MTT(101.89±2.24)s in E group prolonged(P <0.05).Tp,MTT,Peak and AUC decreased in E group compared with HS-RI group(P<0.05).3.The results of biochemical indexes: Compared with the values of ALT(64.61±1.50)U/L and AST(133.31±12.44)U/L in Sham group,the values of ALT and AST in HS-RI group and E group were significantly higher(P<0.05).Compared with the values of ALT(231.68±15.49)U/L and AST(903.29±36.08)U/L in HS-RI group,the values of ALT(153.65±5.89)U/L and AST(621.58±8.58)U/L in group E were significantly lower(P<0.05).4.Pathological results: There was no obvious thickening of liver tissue capsule in Sham group,and hepatocytes were normal and arranged radially around the central vein.Liver tissue capsule thickened in HS-RI group,with hepatocyte necrosis and inflammatory cell infiltration.Compared with HS-RI group,liver tissue capsule thickened slightly in group E,accompanied by a small amount of hepatocyte necrosis and a small amount of inflammatory cell infiltration.5.The results of ELISA detection: Compared with Sham group,the contents of i NOS and IL-6 in HS-RI group and E group increased,while the content of Bcl-2 decreased(P<0.05).Compared with i NOS(24.75±1.00)ng/m L,IL-6(13.59±0.94)pg/m L and Bcl-2(25.60±1.19)ng/m L in HS-RI group,i NOS(19.86 ± 1.47)ng/m L and IL-6(10.34 ± 0.60)pg/m L content decreased and Bcl-2(34.22 ± 1.12)ng/m L content increased in E group(P<0.05).6.Correlation analysis of VTIQ parameters with biochemical and ELISA indexes: SWVmin,SWVmax,SWVmean were significantly correlated with biochemical and ELISA indexes(P<0.001),and SWVmean was highly correlated with ALT,AST,Bcl-2,IL-6 and i NOS(r=0.872;0.846;-0.857;0.828;0.865).7.Correlation analysis between contrast-enhanced ultrasound parameters and biochemical and ELISA indexes: Except for the correlation between Peak and AST,the correlation between contrast-enhanced ultrasound parameters and biochemical and ELISA indexes was statistically significant(P<0.05).AUC and MTT were highly correlated with ALT,AST,Bcl-2,IL-6 and i NOS,and r|were(0.928;0.896;-0.954;0.916;0.862)and(0.970;0.973;-0.978;0.948;0.863),respectively.Conclusion:1.VTIQ technique can evaluate the changes of liver tissue hardness during resuscitation of hemorrhagic shock and evaluate the effect of prophylactic medication.SWVmin,SWVmax and SWVmean all have certain reference value.2.CEUS can monitor the microcirculatory perfusion abnormality of acute liver injury in resuscitation stage of hemorrhagic shock,and can find the perfusion changes of acute liver injury after estradiol intervention,in which Tp,AUC and MTT have certain reference value.3.The quantitative parameters of VTIQ and CEUS are significantly correlated with biochemical and ELISA indexes,and their changes could reflect the degree of liver HS-RI.4.VTIQ and CEUS techniques provide a new and sensitive detection method for detection of acute liver injury in resuscitation stage of hemorrhagic shock and evaluation of curative effect after drug intervention. |
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