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Objective: ultrasound contrast (Contrast-enhanced Ultrasound, CEUS) to quantify the quantitative assessment rabbit kidney ischemia-reperfusion injury, and compare the parameters of the renal cortex time - intensity curve (time intensity curve, TIC) and its renal tissue ICAM-1 (Intercellular Adhesion Molecule-1, ICAM-1) expression levels, contrast-enhanced ultrasound in renal ischemia-reperfusion injury diagnostic value. Methods: 24 healthy New Zealand rabbits were randomized divided into a control group (I0), in accordance with the different point in time, ischemia-reperfusion group, which at different reperfusion time is divided into 24 h of reperfusion group (I24h) again perfusion 3 d group (I3d), reperfusion 5'd group (I5d) 4 groups of 6 each. Preoperative cut the rabbit kidney District angora, observe the rabbit kidney position and mark with a two-dimensional gray-scale ultrasound intramuscular injection followed by rabbit leg muscles most abundant at Sumianxin Ⅱ general anesthesia for kidney area cotton ball with alcohol aseptic technique after local skin disinfection. Control group, only the removal of the right kidney, left kidney not treated, the abdomen was closed. Ultrasound examination. 18 I24h, I3d, I5d3 group rabbit line right kidney resection, separation of the left kidney, noninvasive arterial clamp the renal pedicle occlusion of the left kidney blood vessels caused by ischemia, observed the rapid change from dark red kidney bright red, indicating lack of successful blood start ischemia time. The artery clip one hour after the release, the kidney changed from dark red to bright red, reperfusion instantly start ischemia-reperfusion. The abdomen was closed room Wenqing Jie feeding. , Respectively, at 24 h, 3 d, 5 d different time points including two-dimensional, color Doppler (color the Doppler flow imaging, CDFI), power Doppler (power Doppler imaging, PDI) and contrast-enhanced ultrasound (CEUS), including ultrasound examination. Compare different check state renal blood perfusion. Echo ultrasound contrast mode and quantitative analysis software (Wash in / Wash out) in different parts of the renal cortex obtained in the analysis of harmonic state time - intensity curve (TIC). Parameters include start with enhanced time (arrival time, AT), the perfusion peak time (time to peak, the TTP), the perfusion peak intensity variation (difference between B and the Intensity at T = 0, A), the area under the curve (Area under the curve ascending branch, Area), curve slope (gradient between start frame to peak frame, Grad). White rabbits were sacrificed immediately after each set of ultrasound contrast detection and biopsy of renal cortex tissue was taken under low temperature. A portion of refrigerated at -60 ℃, based on double antibody sandwich technology, in strict accordance with the enzyme-linked immunosorbent assay kit measured each group rabbit renal cortex tissue cell adhesion molecule-1 expression in the content and analysis with ultrasound contrast TIC curve of each parameter correlation. Between the two groups, the group within the parameters of the single-factor analysis of variance was used to compare the homogeneity of analysis test, different groups of TIC curve parameters corresponding ICAM-1 expression in renal tissue content relationship using linear correlation analysis, calculate the correlation coefficient r value to P lt ; 0.05 was considered statistically significant. Results: experiments rabbit anesthesia markedly good modeling smooth, angiography were fully awake state. (1) conventional two-dimensional ultrasound the rabbit renal morphology rules from the surface depth of about 1.3-1.8cm, about the size of (2.85 ± 1.03) x (2.23 ± 0.67) cm2, encapsulated, real echo uniform, CDFI and PDI able to display the rabbit kidney was the dendritic distribution vascular branch. And pulsed Doppler (Pulsed wave Doppler, PW) can detect a rabbit renal artery blood flow velocity change, but the CDFI not clearly show the arcuate arteries its artery perfusion, PDI subcapsular perfusion , but quantization scale indicators. (2) the results of ultrasound contrast harmonic state: (1) control group: the marginal ear vein bolus injection of contrast agent SonoVue 3-8s, the contrast agent quickly from the renal sinus of renal subcapsular renal segment artery, interlobar artery arcuate artery, renal cortex, renal medulla, renal sinus turn strengthen, to the entire left kidney formation the fireball like \(2) ischemia-reperfusion group: Enhanced mode with the naked eye began to increase at the time and subsided time longer, but not very different. (3) TIC curve parameters were ① I0 groups: the renal cortex TIC curve performance for a rapid decline after the rapid rise up to the peak, the parameters are as follows: A (15.027 ± 13.488dB), AT (8.628 ± 1.546s) TTP (4.671 ± 0.853s ). Area (630.573 ± 137.793dBs) Grad (4.176 ± 0.2911 dB / s); ② 124h group: TIC curve showed a slow rise, slow the rate of decline, the parameters are as follows: A (16.626 ± 15.271dB), AT (12.806 ± 4.011s) TTP (18.187 ± 4.172s) Area (971.799 ± 206.877dBs) Grad (1.498 ± 1.460 dB / s); the ③ I3d groups: TIC curve rising slower rate of decline is slower, the parameters are as follows: A rise in (15.846 ± 16.233dB) AT (13.492 ± 4.978s) TTP (23.692 ± 7.232s). Area (939.190 ± 393.443dBs). Grad (1.196 ± = 0.977, respectively dB / s); ④ I5d group: TIC curve performance the speed than I24h the group and I3d group, but less than I0 group, the rate of decline is also the same. The parameters are as follows: A (18.655 ± 15.835dB). AT (10.567 ± 3.849s). TTP (13.313 ± 3.913s). Area (964.248 ± 300.197dBs), Grad (2.014 ± 0.474 dB / s). Between the two groups found that: consistent: compare trends between ① the TTP and AT values ??group and reperfusion group than in the control group to extend the reperfusion 24h reperfusion 3d when showing a rising trend in the third Telsda peak reperfusion 3d to reperfusion first 5d was decreasing, but reperfusion the 5d still longer time than the normal control group, the difference was statistically significant (P lt; 0.05). ② Grad values ??among the three groups, the difference was significant (P lt; 0.01): ischemia-reperfusion group compared with the control group slope were significantly lower, and the difference between the reperfusion group is also very significant. Ischemia-reperfusion the 3d curve Grad values ??lowest rises slowly rebounded reperfusion 5d rise accelerated performance, but still slower than the normal control group; ③ Area values ??ischemia reperfusion group was significantly higher than the control group (P lt; 0.05 ), but between ischemia-reperfusion group had no significant difference (P gt; 0.05). ④ A value between the groups, no significant change (P gt; 0.05), not statistically significant. (4) The results of pathological examination: normal control group nephron clear, the tubular glomerular neatly; 24 h of reperfusion group, rabbit renal tubular epithelial cells increase in size, to vacuolization, ballooning degeneration Lord, interstitial lymphocytes was significantly increased inflammation infiltration; reperfusion 3d group the rabbit kidney tubular lumen expansion, part of the epithelial cells of the phenomenon of nuclear enrichment, nuclear fragmentation and nuclear dissolved, was coagulation necrosis, necrotic area surrounding congestive and inflammatory response with interstitial edema is obvious, visible a large number of infiltration of lymphocytes; reperfusion 5d groups, the expansion of the tubular lumen, lumen seen fewer cell tube type, see only a small number of epithelial cells degeneration and necrosis, newborn capillary an increase in the number of blood vessels, interstitial edema, lymphocytes. (5) the concentration of the standard, and the corresponding OD values ??calculated linear regression equation of the standard curve, and then according to the OD value of the sample to calculate the corresponding concentration of the sample in the regression equation. The renal cortex ICAM-1 expression levels following results: I0 group (29.272 ± 5.257ng/ml), I24h group (58.770 ± 6.599ng/ml), I3d group (80.120 + 10.539ng/ml) I5d group (50.676 ± 6.319ng / ml). Content of ischemia-reperfusion group compared with the control group were significantly increased P lt; 0.05, the difference was statistically significant. Ischemia-reperfusion group compared I3d group was the highest the, I5d group declined, although below I24h group but still higher than that in control group, a significant difference (P lt; 0.01) and group comparisons were statistically significant. (6) The the TIC curve parameters and. Linear correlation analysis of the renal cortex ICAM-1 expression levels: Grad with kidney cortex ICAM-1 expression levels showed a negative correlation (correlation coefficient r = -0.923, P lt; 0.01), AT, TTP and renal cortical expression of ICAM-1 content There was a positive correlation (related coefficient r, respectively, for r = 0.697, r = 0.892, P lt; 0.01), A, Area value and the expression of ICAM-1 expression content no correlation (correlation coefficient r = 0.108, r = 0.295, P gt; 0.05) Conclusion: (1) ultrasound imaging techniques than conventional ultrasound is more sensitive to reflect rabbit renal ischemia-reperfusion injury around the the renal cortex tiny artery perfusion status; (2) ultrasound contrast the TIC curve can dynamically reflect the blood The contrast agent concentration changes over time, the obtained quantitative parameters the microbubbles filling speed and balance microvesicle acoustic strength, i.e., to accurately reflect the erythrocyte volume in microvascular flow velocity of the blood flow and tissue; (3) rabbit renal ischemia reperfusion injury occurred three days after a large number of neutrophil adhesion to endothelial cells and tissue spaces, resulting in a severe inflammatory reaction, and at the same time produce large amounts of necrotic material blocking the vascular lumen, renal tissue under the most severe injury. Tide over the dangerous period, renal function can be partially or fully restored. Further elaborated neutrophil-mediated mechanism of ischemia-reperfusion injury. (4) between the the TIC curve parameters and renal tissue ICAM-1 expression content that ultrasound contrast detection technology not only quantitative assessment of renal perfusion state, but also non-invasive reflect kidney tissue inflammation extent and scope; ( 5) ultrasound imaging is non-invasive, safe and reliable inspection method, quantitative assessment of regional renal blood flow and overall, is expected to be a simple, non-invasive detection of sensitive reflects renal ischemia-reperfusion injury nephritis disease reaction method. Clinical early discovery, early diagnosis of renal ischemia reperfusion injury related to the disease and its prognosis and efficacy evaluation of new methods and new ideas.
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