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ydsc526銅蟲(chóng) (初入文壇)
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[求助]
藥物滲透性小鼠模型
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各位高手們，我要做下面的實(shí)驗(yàn)，下面兩段是我摘的兩篇文獻(xiàn)上的片段 2.1 藥物滲透模型 首先對(duì)動(dòng)物背部進(jìn)行脫毛，然后麻醉，一般采用戊巴比妥麻醉動(dòng)物（30mg/ kg），麻醉成功后用砂紙對(duì)動(dòng)物背部進(jìn)行打磨，用合適大小的小桶粘附與動(dòng)物背部打磨部位，接著在小筒內(nèi)進(jìn)行給藥。小筒一般直徑1～2cm即可，要注意打磨程度。一般可在給藥后1h、2h、4h進(jìn)行檢測(cè)[3]。藥物可通過(guò)高效液相色譜法（HPLC）和免疫印跡法（western blot）進(jìn)行檢測(cè)�？捎^察得到藥物透皮能力，監(jiān)測(cè)藥物的有效作用。此模型適用于大鼠、小鼠等動(dòng)物。 The mice were anesthetized via intramuscular injections of Zoletil 50 at a dosage of 50 mg/kg of body weight. In order to assess the skin permeability of HG1, the shaved back skin of each experimental mouse was scratched with No. 600 sandpaper. A plastic cylinder with a diameter of 1.65 cm was affixed to the abrasion site using cyanoacrylate adhesive. A 200 ul volume of 1% (wt/vol) HG1 solution (10 mM sodium phosphate [NaP] buffer [pH 7.4] [10%], polyethylene glycol 400 [45%], glycerol [45%]) was poured into the cylinder chamber. The cylinder was removed at a predetermined time, after which the skin was washed extensively with NaP buffer (pH 7.4) to completely remove any HG1 that might remain on the surface of the skin. An area of skin (1-by-1 cm) was then excised and laid out on a slide glass. The skin samples were then frozen at - 80°C for 1 h. A constant volume of tissue was obtained from each of the frozen skin samples by the use of a biopsy punch with a diameter of 8 mm. The isolated tissues were homogenized in liquid nitrogen, and the proteins were extracted from the homogenates via overnight shaking in 0.5 ml of 5% acetic acid. After brief centrifugation, 0.1 ml of the supernatant was subjected to C18 reversed-phase high-performance liquid chromatography (HPLC) 我按照這篇英文文獻(xiàn)上的步驟這么做，但是在離心取上清的時(shí)候遇到問(wèn)題了，因?yàn)樵谂蹾PLC之前樣品要過(guò)0.22um濾膜，但是吸上來(lái)的上清是膠狀的，根本過(guò)不了濾膜，所以請(qǐng)各位幫我分析一下，謝謝了。 |
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