缺氧肾脏损伤研究

实验研究

研究采用30只5周龄SD大鼠，进行不同条件的缺氧造模。

传统上，对于慢性缺氧模型，大鼠接受2周缺氧(10% O2)。氧合目标(Pa O2 90-60mmHg)是通过调整吸气氧气的分数(Fi O2)来实现的，对于高生存率和低PaO2水平，研究人员在研究中测量的最小O2浓度为7% O2。老鼠随机分布成三组根据吸入不同浓度的氧气（每组包括10只动物）：21% O2在B组，10% O2和7% O2组每组进一步分为两个子组根据缺氧时间（1或2周）（图1）

       对照组大鼠暴露于21%的O2，而实验组大鼠暴露于不同程度的缺氧至少1周。缺氧组大鼠置于常压低氧舱(ProOx-100，上海塔望科技，中国)。对照常氧组暴露在室内空气中，缺氧组被放置在缺氧室(每笼n=5)。氧浓度范围和准确性分别为0-25%和0.1%。

*如上图，塔望科技可提供

       低氧结束，大鼠注射麻醉。麻醉后，将大鼠仰卧位，固定在手术台上。采用腹侧中线切口隔离腹主动脉，将聚乙烯导管插入腹主动脉，并连接压力传感器进行数据采集血压测量系统。通过腹主动脉测量血压，并采血进行检测。

*如上图，塔望科技可提供

       然后对这些大鼠进行放血安乐死。切除双肾进行分析。其中一个标本被闪存冷冻，立即保存在液氮容器中进行免疫印迹分析，而另一个样品在4%多聚甲醛溶液中浸泡进行纤维固定，然后进行染色和免疫组化检查。在尼康EclipseE600显微镜下观察组织切片，并使用尼康DS-Ri1数码相机拍摄不重叠的图像。

部分结果如下：

FIGURE 3 | The growth of rats and the weight of rat kidneys. (A) The mean body weight of the rats was determined on a daily basis for 14 consecutive days (10 rats were included in the analysis for the fifirst 7 days, and 5 rats were included for the remaining 7 days). (B) The renal hypertrophy index of rats on D7 (n = 5) and D14 (n = 5). (C) The mean left kidney weight of the rats was determined on D7 (n = 5) and D14 (n = 5). (D) The mean left kidney weight of the rats was measured on D7 (n = 5) and D14 (n = 5).

FIGURE 4 | General features and morphological traits in renal tissues obtained from animals subjected to normoxic and hypoxic conditions. (A) Gross view of a kidney. (B) A histological section of kidney tissue. The histological examination was performed at 20 × magnifification (scale bar = 100 µm).

(C) Periodic acid-Schiff staining (40 × magnifification, scale bar = 50 µm).

(D) Masson’s trichrome staining (40× magnifification, scale bar = 50 µm).