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用地塞米松构建建鲤肝切片脂变模型,并探索最佳建模浓度,初步探究地塞米松致建鲤肝脏脂变的作用机理。建鲤活体取肝,用振荡切片机制备厚度为300μm的建鲤肝切片,27℃培养箱中预培养2 h。将试验分为试验组和空白对照组,试验组用含浓度为0.039 3、0.393 0、3.930 0、39.300 0、393.000 0 mg/L地塞米松的L-15培养基于27℃培养箱中培养12 h,空白对照组用L-15培养基培养相同时间,培养结束后,收集试验组和空白对照组中上清液和肝切片。参照试剂盒说明检测上清液中谷丙转氨酶(GPT)、谷草转氨酶(GOT)活性,肝切片匀浆中三磷酸腺(ATP)、总蛋白(TP)含量,甘油三酯(TG)、总胆固醇(TC)、游离脂肪酸(FFA)含量以及脂肪酸合成酶(FAS)活性。结果显示,建鲤肝切片经地塞米松诱导后,ATP含量变化不大;当地塞米松浓度为3.93 mg/L时,试验组的其他指标与空白对照组差异极显著(P<0.01)。由此说明,用浓度为3.93 mg/L地塞米松诱导建鲤肝切片12 h可以成功构建建鲤肝切片脂变模型。
Constructed the dexamethasone to build the carp liver fragment lipid model, and explored the best model concentration, to explore the mechanism of dexamethasone induced carotid liver fat. Jianli live take liver, using a vibrating microtome machine to prepare Jian carp liver slices thickness of 300μm, 27 ℃ incubator pre-culture 2 h. The experiment was divided into experimental group and blank control group. The experimental group was cultured in L-15 medium containing dexamethasone with concentration of 0.039 3,0.393 0,3.930 0,39.300 0,393.000 0 mg / L in a 27 ° C incubator h, blank control group cultured with L-15 medium for the same time, after the end of culture, the supernatant and liver slices of the experimental group and the blank control group were collected. Reference to the kit instructions to detect the supernatant of alanine aminotransferase (GPT), aspartate aminotransferase (GOT) activity, hepatic slices homogenate in the triphosphate (ATP), total protein (TP) content, triglyceride (TG), total cholesterol (TC), free fatty acid (FFA) content and fatty acid synthase (FAS) activity. The results showed that there was no significant change of ATP content in Jianshi liver slices induced by dexamethasone; when the concentration of dexamethasone was 3.93 mg / L, the other indexes of the experimental group were significantly different from the blank control group (P <0.01). The results showed that the hepatic sections of Jian carp could be successfully constructed by using the concentration of 3.93 mg / L dexamethasone to induce the liver slices of Jian carp for 12 h.