褐藻多糖硫酸酯对 TGF-β1体外诱导的肾小管上皮细胞间质转分化过程中细胞骨架的影响

目的：通过体外观察褐藻多糖硫酸酯对TGF-β1诱导肾小管上皮细胞（HK-2）间质转分化过程中细胞角蛋白（cytok-eratin，CK）和α-平滑肌肌动蛋白（α-smooth muscle action ，α-SMA）表达的影响，探讨该药对HK-2上皮-间质转分化的影响及其作用机制。方法体外培养正常人肾小管上皮细胞（ HK-2）细胞株，采用倒置相差显微镜观察对照组、诱导组和药物组细胞形态的变化，免疫细胞化学SABC法和图像分析技术检测培养第3 d对照组、诱导组和药物组HK-2细胞CK和α-SMA阳性细胞的平均光密度值。结果培养第3 d，对照组HK-2细胞呈现铺路石样上皮细胞形态特点，诱导组细胞呈长梭形，类似纤维细胞，药物组细胞以上皮细胞为主，少数细胞呈梭形；与对照组相比较，诱导组CK阳性细胞平均光密度值显著降低（ P＜0.05），α-SMA阳性细胞平均光密度值增加（P＜0.05）；与诱导组相比较，药物组CK阳性细胞平均光密度值显著增加（P＜0.05），α-SMA阳性细胞平均光密度值显著降低（P＜0.05）。结论①褐藻多糖硫酸酯具有抑制TGF-β1体外诱导的肾小管上皮细胞间质转分化的作用；②褐藻多糖硫酸酯可能通过改变细胞骨架成分发挥抑制肾小管上皮细胞间质转分化作用。

Effects of Fucoidan on the cytoskeleton during the course of epithelial mesenchymal transition induced by TGF-β1 of human renal tubular epithelial cell in vitro

Objective To observe the Fucoidan in vitro against TGF-β1-induced renal tubular epithelial cells （ HK-2 ） mesenchy-mal transition process cytokeratin （cytokeratin,CK） and α-smooth muscle actin （α-smooth muscle action,α-SMA） expression,and to explore the Fucoidan on HK-2 epithelial-mesenchymal transition and its mechanism .Methods We cultured cell lines of normal human renal tubular epithelial cells （HK-2）.Using an inverted phase contrast microscope ,we observed morphological changes of cells in con-trol group,induction group and drug group .With the use of immune cells SABC method we cultured HK-2 cells CK andα-SMA-positive cells of the control group ,the induction group and the drug group for 3 days and detected their average optical density values with image analysis .Results After three days of culture ,HK-2 cells showed cobblestone morphology characteristic of epithelial cells in the control group,long fusiform in the induction group ,which was similar to fibroblasts,and mainly epithelial cells over cells in the drug group ,with a few cells fusiform.Compared with the control group ,the average optical density values of CK-positive cells in the induced group were significantly lower （P〈0.05）,which those of α-SMA-positive cells increased （P〈0.05）.Compared with the induction group the av-erage optical density values of CK-positive cells in the drug group were significantly incerased （ P〈0.05 ） , Wile those of α-SMA-posi-tive cells were significantly lower （P〈0.05）.Conclusion ①Fucoidan inhibits vitro mesenchymal transdifferentiation of TGF-β1-in-duced renal tubular epithelial cells;②Fucoidan may play a part inhibiting mesenchymal transdifferentiation of renal tubular epithelial cells by altering the cytokeleton .