實(shí)驗(yàn)研究

研究采用30只5周齡SD大鼠，進(jìn)行不同條件的缺氧造模。

傳統(tǒng)上，對于慢性缺氧模型，大鼠接受2周缺氧(10% O2)。氧合目標(biāo)(Pa O2 90-60mmHg)是通過調(diào)整吸氣氧氣的分?jǐn)?shù)(Fi O2)來實(shí)現(xiàn)的，對于高生存率和低PaO2水平，研究人員在研究中測量的最小O2濃度為7% O2。老鼠隨機(jī)分布成三組根據(jù)吸入不同濃度的氧氣（每組包括10只動物）：21% O2在B組，10% O2和7% O2組每組進(jìn)一步分為兩個子組根據(jù)缺氧時間（1或2周）（圖1）

? ? ? ?對照組大鼠暴露于21%的O2，而實(shí)驗(yàn)組大鼠暴露于不同程度的缺氧至少1周。缺氧組大鼠置于常壓低氧艙(ProOx-100，上海塔望科技，中國)。對照常氧組暴露在室內(nèi)空氣中，缺氧組被放置在缺氧室(每籠n=5)。氧濃度范圍和準(zhǔn)確性分別為0-25%和0.1%。

*如上圖，塔望科技可提供

? ? ? ?低氧結(jié)束，大鼠注射麻醉。麻醉后，將大鼠仰臥位，固定在手術(shù)臺上。采用腹側(cè)中線切口隔離腹主動脈，將聚乙烯導(dǎo)管插入腹主動脈，并連接壓力傳感器進(jìn)行數(shù)據(jù)采集血壓測量系統(tǒng)。通過腹主動脈測量血壓，并采血進(jìn)行檢測。

*如上圖，塔望科技可提供

? ? ? ?然后對這些大鼠進(jìn)行放血安樂死。切除雙腎進(jìn)行分析。其中一個標(biāo)本被閃存冷凍，立即保存在液氮容器中進(jìn)行免疫印跡分析，而另一個樣品在4%多聚甲醛溶液中浸泡進(jìn)行纖維固定，然后進(jìn)行染色和免疫組化檢查。在尼康EclipseE600顯微鏡下觀察組織切片，并使用尼康DS-Ri1數(shù)碼相機(jī)拍攝不重疊的圖像。

部分結(jié)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).