| 摘要: |
| 目的：结合前期微泡增强的高声压脉冲超声空化作用致兔肝血流灌注降低的研究结果，进一步探讨高声压超声辐照对兔肝细胞水通道蛋白4（Aquaporin 4, AQP 4）及钠钾ATP酶（Na+-K+ ATPase）表达的影响作用，通过检测不同辐照时间作用后AQP 4及Na+-K+ ATPase蛋白表达量的改变，从而探究水分子转运相关蛋白表达与肝细胞肿胀之间的关系。方法：健康新西兰大白兔41只，随机分入空白对照组（n=5）及不同辐照时间组：T1组（辐照时间1 min，n=12）、T5组（辐照时间5 min，n=12）及T10组（辐照时间10 min，n=12）。T1、T5及T10组动物采用高峰值负压（P=2.0 MPa）超声脉冲联合经静脉脂质微泡注射进行肝脏超声空化辐照处理，超声辐照后获取肝脏组织标本；空白对照组动物仅行开腹手术获取肝脏组织标本。免疫组织化学法观察各组AQP 4及Na+-K+ ATPase蛋白表达分布情况，蛋白免疫印迹法（WB）检测各组AQP 4及Na+-K+ ATPase蛋白表达量。结果：超声辐照治疗后各治疗组AQP 4及Na+-K+ ATPase强阳性表达细胞较空白对照组数量增多、分布范围广泛；T1、T5及T10组AQP 4蛋白表达量较对照组均显著增高，差异有统计学意义（P＜0.05），组间比较提示T1、T10组与T5组AQP 4蛋白表达量差异均有统计学意义（P＜0.05），而T1组和T10组之间AQP 4蛋白表达量差异无统计学意义（P＞0.05）；T5组及T10组Na+-K+ ATPase蛋白表达量明显高于空白对照组及T1组，差异均有统计学意义（P＜0.05），两组之间比较其差异亦有统计学意义（P＜0.05），而T1组与空白对照组比较差异没有统计学意义（P＞0.05）。 结论：高声压脉冲超声空化作用在引起正常兔肝组织水肿的同时可引起AQP 4及Na+-K+ ATPase蛋白表达量的增加，超声辐照时间的长短对蛋白表达量的增加有影响。 |
| 关键词: 超声空化 微泡 肝脏 水通道蛋白 钠钾ATP酶 |
| DOI： |
| 投稿时间：2020-11-06修订日期：2020-12-01 |
| 基金项目:国家自然科学基金项目（编号81301239），湖北省卫生健康科研基金项目（编号WJ2019H091） |
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| Effects of acoustic cavitation on the expression of aquaporin 4 and sodium potassium ATP enzyme in rabbit hepatocytes |
| gaoyuejuan,wangfei,dongxiaoxiao,gaowenhong |
| (Department of ultrasound, the 5th center of PLA general hospital) |
| Abstract: |
| Objective: Microbubble enhanced cavitation of high negative pressure pulsed ultrasound has been proved the ability of causing swollen hepatocytes and decreasing hepatic blood perfusion of rabbit. In this study, the relationship between microbubble enhanced cavitation and two kinds of water molecular transport related proteins-aquaporin 4 (AQP 4) and sodium potassium ATP enzyme (Na+-K+ ATPase)-were further investigated. Methods: Forty-one healthy New Zealand white rabbits were randomly divided into the control group (n=5) and three different irradiation time groups: group T1 (irradiation time=1 min, n=12), group T5 (irradiation time=5 min, n=12) and group T10 (irradiation time=10 min, n=12). Animals in group T1, group T5 and group T10 were treated with high peak negative pressure (P=2.0 MPa) ultrasound irradiation combined with microbubble. Liver tissue specimens were obtained after ultrasound irradiation. Liver tissue specimens were taken by laparotomy in the control group. Distribution of AQP 4 and Na+-K+ ATPase in each group was observed by immunohistochemical method, and expression of both protein was detected by western blot. Results: IHC results shows the number of AQP4 and Na+-K+ ATPase positive cells in treatment groups is much higher than that in control group. AQP4 protein expression in T1, T5 and T10 group is significantly higher than that in control group (P < 0.05). Comparison between groups shows that AQP4 protein expression in T5 group is significantly higher than that in T1 and T10 group (P < 0.05), while the difference between T1 and T10 group is not statistically significant. Na+-K+ ATPase in T5 group and T10 group is significantly higher than that in control group and T1 group (P < 0.05), while no significant difference shows between T1 group and control group (P > 0.05). Expression of Na+-K+ ATPase in T5 group is also significantly higher than that in T10 group (P < 0.05). Conclusion: Microbubble enhanced cavitation could cause increased expression of AQP 4 and Na+-K+ ATPase in normal rabbit liver while inducing liver edema. Ultrasonic irradiation time had an effect on the increase of protein expression. |
| Key words: Cavitation Microbubble Liver Aquaporin Sodium potassium ATP enzyme |
