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Resesrch On Contrast-enhanced Ultrasound To Evaluate Testicular Ischemia-Reperfusion Injury And The Protective Effects Of Matrix Metalloproteinases Inhibitor

Posted on:2017-08-11Degree:MasterType:Thesis
Country:ChinaCandidate:Q F QianFull Text:PDF
GTID:2334330503473678Subject:Medical imaging and nuclear medicine
Abstract/Summary:PDF Full Text Request
ObjectiveTo explore the protective effects and the correlation mechanism of exogenous matrix metalloproteinases inhibitor on rabbit testicular moderate ischemia/reperfusion injury. Observe the TMB-9 ultrasound contrast performance before and after the intervention by doxycycline.Investigate an objective and noninvasive method to evaluate the testicular microvascular injury.MethodsA total of 36 rabbits were randomly divided into 6 groups including control groups(Group S1 and Group S2), ischemia reperfusion groups(Group R1 and Group R2) and doxycycline intervention groups(Group D1 and Group D2) with 6 in each group. The testis on the one side was randomly selected and a longitudinal incision was made above the root of the scrotum approximately 2cm from the upper pole of testis. After the blunt separation and exposure of the structure of the spermatic cord, the cord was completely ligated. Under the monitoring of color Doppler ultrasound, the blood flow signal in the testis of the operation side was lost, which indicated the establishment of a complete ischemia model. The reperfusion was performed on the testis of the operation side 4h after complete ischemia, followed by 1-day feeding for Group R1 and D1 as well as 4-day feeding for Group R2 and D2. In the control groups(Group S1 and Group S2), threading without ligation was performed after the exposure of the spermatic cord. The doxycycline intervention groups(Group D1 and Group D2) and Group S1 were given intraperitoneal injection of doxycycline 20mg/(kg.d). The ischemia reperfusion groups(Group R1 and Group R2) and Group S2 were given intraperitoneal injection of equivalent normal saline.Before operation, MB and TMB-9 contrast-enhanced ultrasound(CEUS) was performed in each group after the successful establishment of each model to analyze the changes of contrast parameters of each group with the ultrasound contrast quantitative analysis technologies including PI, TP, Slope, MTT, DT/2 and AUC and to compare the two evaluation results. After radiography, 3 ml heart blood was obtained by puncture in each group. The plasma NO, ET-1 and v WF contents were detected in the laboratory. HE staining and immunohistochemistry staining were performed on the testis of the operation side after radiography to observe the pathological changes and the changes of MMP-9 expression of each group. SPSS17.0 software was used for statistical analysis.ResultsLipid microbubbles is milky white turbid liquid. The diameter of the microbubble size is uniform, about 3.0 to 6.0?m. The number of microbubbles was decreased under the condition of room temperature for 30 minutes.The numble of microbubbles was decreased significantly and the diameter of microbubbles would be inconsistent when stored at room temperature 24 hours later.There was no significant difference in preoperative MB and TMB-9 parameters of CEUS among each group.The MB ultrasound contrast parameters including PI, AUC, MTT and DT/2 of the Group R1 significantly increased, while the parameter TP significantly decreased(P<0.05). And the changes of the contrast parameters of TMB-9 were much more obvious.The parameters including PI, AUC, MTT and DT/2 of the Group R1 significantly increased, and the parameter TP significantly decreased(P<0.01),and the parameter Slope meanwhile increased(P<0.05). PI of MB ultrasound contrast parameters of the Group R2 decreased(P<0.05), and the other parameters had no significant differences(P>0.05). While compared with these of the Group R1(P<0.05), TMB-9 contrast parameters of Group R2 including PI and Slop decreased, and the differences of other parameters were not statistically significant(P>0.05)In the doxycycline intervention Group D1, MB and TMB-9 contrast parameters including PI, MTT, DT/2 and AUC decreased(P<0.05), compared with these of Group R1. Four days after reperfusion, compared with these of Group R2, MB and TMB-9 contrast parameters including including PI, MTT, DT/2 and AUC significantly decreased(P<0.01). Compared with these of the control group, all the MB contrast parameters had no significant differences(P>0.05). In TMB-9 contrast parameters, only PI increased(P<0.01), while no significant difference was found in other contrast parameters.There was no significant difference in plasma v WF, NO and ET-1 levels between each group before operation(P>0.05). Compared with these of group S2, plasma VWF, ET-1 and NO contents increased significantly in Group R1 after operation(P<0.01). Compared with these of Group R1, plasma v WF and ET-1 content of Group R2 further increased(P<0.01) while the content of NO had no significant difference(P>0.05). The plasma v WF, ET-1 and NO contents of Group D1 were significantly higher than these of the Group S1(P<0.01). Compared with these of Group D1, the plasma v WF content of Group D2 further increased(P>0.05), while ET-1 and NO contents had no significant difference(P>0.05).Results of pathological examination showed that the microvascular structure of interstitial substance of testis of Group S1 and S2 was complete and clear without dilatation and congestion. The microvessels of interstitial substance of testis of ischemia reperfusion Group had congestion and hemorrhage with an obscure structure. A large number of leakage of red blood cells and infiltration of inflammatory cells were found in the surrounding interstitium and partial continuous interrupt could be found in microvessels in Group R1. Immunohistochemical staining showed that MMP-9 was largely expressed in the microvascular basement membrane of testis. Compared with the ischemia reperfusion group, the expression of MMP-9 in the vascular basement membrane of doxycycline intervention groups was significantly decreased(P<0.01), which have a much more clearly structure of testicular microvascular basement membrane. The infiltration of inflammatory cells and leakage of red blood cells were significantly subsided in the surrounding interstitium.Conclusions1. Doxycycline can effectively inhibit testicular moderate ischemia-reperfusion injury microvascular basement membrane expression of MMP-9, which helps to maintain the integrity of microvascular basement membrane and the surrounding stroma,and reduce the injury.2. TMB-9 can be more sensitive and objective to evaluating the injury of testicular microvascular basement membrane than MB.
Keywords/Search Tags:testicular microvascular, ischemia-reperfusion injury, MMP-9, doxycycline, ultrasound targeted contrast imaging
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