| Objective:To establish the rabbit model of myocardial ischemia reperfusion by with interleukin 8 (IL-8) monoclonal antibody line acoustic contrast myocardial acoustic imaging to carry out targeted research.Methods:1.preparation and determine the rabbit model of myocardial ischemia reperfusion:65 of Japanese big ear rabbits were randomly divided into two groups:Myocardial ischemia reperfusion (I/R group) and control group (Sham group),I/R group of homemade blocking devices used to open the chest to block left anterior descending coronary artery blood flow, and in 90 min of ischemia lifting block, reperfusion began;After the Sham group to open the chest does not block the left anterior descending coronary artery blood flow, is consistent with the rest of the I/R group.Intraoperative continuous ecg monitoring,In two groups of rabbit, respectively, 90 min of ischemia and reperfusion for 60 min,120 min,180 min and 1 w via artery in rabbit ear extraction 3 ml of blood samples, determination of troponin I (cTnI), creatine kinase MB isoenzyme (CK-MB), myoglobin (MB) concentration.Respectively for the above period of time executed after the rabbit heart, line HE staining and MASSON staining to observe the myocardial pathology change, using the immunohistochemical staining experiment on myocardial ischemia reperfusion group monitoring and expression of IL-8.2.targeted acoustic contrast agent preparation and identification:Covalent coupling method is used by SPDP crosslinking agent IL-8 monoclonal antibody coupling in the surface of SonoVue microbubble, microscope micro bubble size, shape and dispersion and through the glass agglutination experiment and fluorescence dyeing experiments show coupling was successful.3.myocardial acoustic imaging inspection:In two groups of rabbit, respectively,90 min of ischemia and reperfusion for 60 min,120 min and 180 min after undergoing routine cardiac ultrasound and 1 w, and respectively prepared with instant SonoVue contrast agents, with IL-8 monoclonal antibody targeting the contrast agent SonoVue, control group contrast myocardial acoustic imaging examination, dynamic images obtained after offline application QLAB software for analysis.4.Statistical methods:application SPSS 17.0 statistical software, rabbit the physiological indexes obey the normal distribution, with the mean ± standard deviation(±), said the same, the two different processing with paired t test, two different samples are used to deal with the same two independent sample t-test, multiple sets of sample mean compared with single factor analysis of variance.: difference was statistically significant (P< 0.05).Results:1.The rabbit myocardial ischemia-reperfusion model:preoperative observing two groups of rabbit two-dimensional ultrasound, ecg are no exception.Electrocardiogram: I/R group rabbits ischemia 90 minst segment elevation, reperfusion began to ST is broken down,;Sham group of rabbit heart rate is a bit faster, ST segment saw no change;I/R group rabbit two-dimensional ultrasound room wall motion amplitude reduction in the Sham group, Sham group no significant change;Serological tests revealed I/R group rabbit serum myocardial injury markers are significantly higher than Sham group (P< 0.05), peak after 180 min reperfusion,1 w gradually dropped to normal range;The Sham group rabbit preoperative and postoperative serum markers of myocardial injury did not see obvious change;Pathological results showed that the I/R group rabbit left anterior descending branch of blood-supply area of myocardial ischemia, myocardial ischemia rabbit saw no obvious Sham group.Immunohistochemical staining showed that myocardial I/R group cell surface almost brown;Myocardial cell surface did not see the Sham group was obviously coloring, which can be proved that ischemia area produce inflammation, and the damage of myocardial tissue release a large number of IL-8.2.Targeted acoustic contrast agent identification:through glass agglutination experiment with immunofluorescence can draw IL-8 monoclonal antibodies has been successfully coupling to the SonoVue microbubble shell surface.3.myocardial acoustic imaging examination:instant preparation of SonoVue contrast myocardial acoustic imaging, building segmental wall, before the interval of the mean contrast ultrasound, ultrasonic peak were significantly lower than that of non inferior wall infarction segments, after the interval, preoperative and Sham group, the corresponding section of the same section (both P< 0.05);The experimental group (with IL-8 monoclonal antibody coupling acoustic contrast agent SonoVue) myocardial acoustic imaging, building of segmental wall, before the interval average contrast ultrasound, ultrasonic peak acoustic contrast agent (SonoVue) were higher than control group (both P< 0.05).To Flash after the first frame as a starting point:SonoVue contrast myocardial acoustic imaging, building segmental wall, before the interval of the contrast agents tmax is later than the inferior wall infarction segments, after the interval, preoperative and Sham group, the corresponding section of the same section (both P< 0.05);Experimental myocardial acoustic imaging, building segmental wall, before the interval of the contrast agent tmax is earlier than the control group the same segment (both P< 0.05).To start the contrast as a starting point, the same only experimental rabbit, experimental myocardial acoustic imaging, building segmental wall, before the interval myocardial began developing time is earlier than the control group in the same section (both P< 0.05).Use of A negative exponential equation [y (t)= A* (1-exp (-B* t))+C] parameters can be concluded that SonoVue contrast myocardial acoustic imaging, I/R group forming die segmental anterior wall, before the interval of myocardial blood volume (A value), myocardial blood flow velocity (B value) and local myocardial perfusion amount (A*B value) were significantly lower than the infarction after segmental wall and interval (all P< 0.05), anterior wall, before the interval of A, B value, A* B values were significantly lower than that of Sham group corresponding section (both P< 0.05);And with IL-8 monoclonal antibody targeted contrast agent, the experimental group made segmental anterior wall, before the interval value, A* B values were higher than control group in the corresponding section (both P< 0.05), and B values in no significant difference between the two groups (all P> 0.05).Conclusion:1.Real-time myocardial acoustic imaging (SonoVue contrast agents) confirmed that the I/R group of ischemia and reperfusion time building segment myocardial blood volume, myocardial blood flow velocity and volume were significantly lower than the infarction myocardial perfusion segments, Sham group, the corresponding segment;Building segment the mean contrast media video intensity of ultrasonic, ultrasonic peak, peak time segment, the Sham group were significantly lower than the infarction, section of the corresponding building segment contrast tmax segment, the Sham group were significantly lower than the infarction corresponding segment, MCE operation is simple, and easy, can be more sensitive to show myocardial segmental enhancement process, can improve the sensitivity of diagnosis myocardial ischemia reperfusion, and enhance the situation through the myocardial each segment to evaluate the situation of myocardial reperfusion.2.The covalent coupling method is adopted through SPDP crosslinking agent IL-8 monoclonal antibody on the surface of the coupling in SonoVue microbubble method is feasible.Various groups of experimental rabbit trauma after open heart surgery, and accepted within 3 h 5 times left heart imaging examination, sparked contrast agents have found no adverse reactions, this suggests that using with IL-8 monoclonal antibody targeting contrast myocardial acoustic imaging examination is a safe and feasible.3.Inflammatory cytokines IL-8 in the early myocardial ischemia (90 min of ischemia) increased significantly, with carrying IL-8 monoclonal antibody targeting contrast myocardial imaging, IL-8 monoclonal antibody and myocardial ischemia of inflammatory factor, reduce inflammation injury.4.with IL-8 monoclonal antibody targeting contrast myocardial acoustic imaging findings:the experimental group building segment contrast video intensity peak of ultrasonic, ultrasonic average higher than that of control group, control group myocardial imaging time to start, tmax are earlier than the control group, suggesting that targeted contrast agent carrying IL-8 monoclonal antibody and ischemic myocardial release of inflammatory factor of IL-8 caused by the reaction between the existence of the targets. |
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