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目的 研究高密度脂蛋白 3(HDL3 )介导大鼠皮肤成纤维细胞胆固醇 (ch)流出机制。方法 用N 乙酰咪唑修饰HDL ,阻断HDL与细胞受体相互作用 ,观察其对HDL3 介导细胞ch流出的影响。用FITC标记HDL示踪HDL3 在介导细胞ch流出中自身代谢过程。结果 牛血清白蛋白 (对照组 )介导 4.74%细胞ch流出 ,HDL3组和N 乙酰咪唑HDL组分别为 31 .85 %和 8.41 %。FITC HDL3 与细胞 37℃培养 3h后 ,细胞内吞荧光强度(FS)占细胞结合FS的 6 5 .78% ,其中 93.5 %FS存在于TCA可沉淀部分。细胞进一步 37℃培养 2h后 ,细胞释放FS占内吞Fs 80 .77% ,其中 92 .7%FS存在于TCA可沉淀部分。结论 HDL3 通过与细胞受体相互作用 ,介导细胞ch流出。其可能机制是HDL3 通过受体进入胞内 ,在胞内不经历溶酶体分解过程。它接受胞内ch后通过逆向胞饮形式流出胞外。
Objective To study the mechanism of high density lipoprotein 3 (HDL3) -induced cholesterol efflux in rat dermal fibroblasts. Methods HDL was modified with N acetylimidazole to block the interaction between HDL and cell receptor, and to observe its effect on HDL3-mediated cholestasis. HDL3 labeled with FITC-labeled HDL3 mediates its own metabolic processes in chilled out cells. Results Bovine serum albumin (control group) induced ch-efflux in 4.74% of cells, which was 31.85% and 8.41% in HDL3 group and N acetylctide HDL group, respectively. FITC HDL3 and cells incubated at 37 ℃ for 3h, the fluorescence intensity of intracellular fluorescence (FS) accounts for 65.78% of the cells bound to FS, of which 93.5% FS is present in the TCA precipitable part. After the cells were further cultured at 37 ℃ for 2h, the cell release FS accounted for 80.77% of the endocytosis Fs, of which 92.7% FS was present in the TCA-precipitable fraction. Conclusion HDL3 mediates cell ch efflux by interacting with cellular receptors. Its possible mechanism is that HDL3 enters the cell through the receptor and does not undergo intracellular lysosomal decomposition. After it receives intracellular ch, it excreted extracellularly through the form of reverse coccus.