目的 探讨川芎中3个苯酞类成分藁本内酯、洋川芎内酯A和洋川芎内酯I对氧糖剥夺/再灌注诱导MDR1转染犬肾细胞（MDCK-MDR1）损伤的作用及机制。方法 采用氧糖剥夺/再灌注诱导MDCK-MDR1细胞损伤模型，给予藁本内酯、洋川芎内酯A和洋川芎内酯I进行干预，采用MTT法检测细胞存活率；采用试剂盒检测细胞上清液中乳酸脱氢酶（lactate dehydrogenase，LDH）释放率；采用流式细胞术检测细胞凋亡率；采用Western blotting法检测细胞紧密连接蛋白如claudin-5、occludin、ZO-1以及外排蛋白P-糖蛋白（P-glycoprotein，P-gp）和葡萄糖转运蛋白-1（glucose transporter-1，GLUT-1）表达情况；以芍药苷为探针药物，研究藁本内酯、洋川芎内酯A和洋川芎内酯I对芍药苷在氧糖剥夺/再灌注诱导的MDCK-MDR1细胞上跨膜转运的影响。结果 与模型组比较，洋川芎内酯I（80 μg/mL）显著提高MDCK-MDR1细胞存活率（P<0.05）；洋川芎内酯I（40、80 μg/mL）显著抑制细胞上清液中LDH释放率（P<0.05、0.01）；藁本内酯（20、40 μg/mL）和洋川芎内酯I显著抑制细胞凋亡率（P<0.01）；藁本内酯（20 μg/mL）和洋川芎内酯I显著上调claudin-5蛋白表达水平（P<0.05、0.01）；藁本内酯（20、40 μg/mL）、洋川芎内酯A（20 μg/mL）和洋川芎内酯I（40、80 μg/mL）显著上调occludin蛋白表达水平（P<0.01）；藁本内酯（20 μg/mL）和洋川芎内酯I（40 μg/mL）显著上调ZO-1蛋白表达水平（P<0.05、0.01）；藁本内酯、洋川芎内酯A（40、80 μg/mL）和洋川芎内酯I显著下调GLUT-1蛋白表达水平（P<0.01）；藁本内酯（20 μg/mL）、洋川芎内酯A和洋川芎内酯I（20 μg/mL）显著下调P-gp蛋白表达水平（P<0.01）；藁本内酯（20 μg/mL）、洋川芎内酯A（40、80 μg/mL）和洋川芎内酯I显著促进芍药苷跨膜转运（P<0.05、0.01）。结论 藁本内酯、洋川芎内酯A和洋川芎内酯I能够上调紧密连接蛋白表达，抑制氧糖剥夺/再灌注诱导的MDCK-MDR1细胞损伤，促进探针药物的跨膜转运。

Objective To explore effect and mechanism of three phthalide components including ligustilide, senkyunolide A, and senkyunolide I from Ligusticum chuanxiong on oxygen-glucose deprivation/reperfusion-induced MDCK-MDR1 cell injury. Methods MDCK-MDR1 cells injury model was induced by oxygen-glucose deprivation/reperfusion, and ligustilide, senkyunolide A and senkyunolide I were used for intervention. MTT was used to detect cell survival rate; Kit was used to detect lactate dehydrogenase (LDH) release rate in cell supernatant; Flow cytometry was used to detect cell apoptosis rate; Western blotting was used to detect cellular tight junction proteins such as claudin-5, occludin, ZO-1 and efflux protein P-glycoprotein (P-gp) and glucose transporter-1 (GLUT-1) protein expression; Paeoniflorin was used as a probe drug to study effect of ligustilide, senkyunolide A and senkyunolide I on transmembrane transport of paeoniflorin in MDCK-MDR1 cells induced by oxygen-glucose deprivation/reperfusion. Results Compared with model group, senkyunolide I (80 μg/mL) significantly increased survival rate of MDCK-MDR1 cells (P<0.05); Senkyunolide I (40, 80 μg/mL) significantly inhibited LDH release in supernatant of MDCK-MDR1 cells (P<0.05, 0.01); Ligustilide (20, 40 μg/mL) and senkyunolide I significantly inhibited apoptosis rate of MDCK-MDR1 cells (P<0.01); Ligustilide (20 μg/mL) and senkyunolide I significantly up-regulated claudin-5 expression (P<0.05, 0.01); Ligustilide (20, 40 μg/mL), senkyunolide A (20 μg/mL), and senkyunolide I (40, 80 μg/mL) significantly up-regulated occludin expression (P<0.01); Ligustilide (20 μg/mL) and senkyunolide I (40 μg/mL) significantly up-regulated ZO-1 expression (P<0.05, 0.01); Ligustilide, senkyunolide A (40, 80 μg/mL) and senkyunolide I significantly down-regulated GLUT-1 expression (P<0.01); Ligustilide (20 μg/mL), senkyunolide A and senkyunolide I (20 μg/mL) significantly down-regulated P-gp expression (P<0.01); Ligustilide (20 μg/mL), senkyunolide A (40, 80 μg/mL) and senkyunolide I significantly promoted the transmembrane transport of paeoniflorin (P<0.05, 0.01). Conclusion Ligustilide, senkyunolide A and senkyunolide I can up-regulate tight junction proteins expressions, inhibit damage of MDCK-MDR1 cells induced by oxygen-glucose deprivation/reperfusion, and promote the transmembrane transport of probe drugs.

R285.5

国家自然科学基金项目（81760714）；国家自然科学基金项目（82060719）；江西省自然科学基金项目（20171BAB205093）；江西省博士后项目（2017KY08）；江西省卫生厅项目（2016A012）；江西省一流学科项目（JXSYLXK-ZHYAO091）；江西省博士启动基金项目（2018WBZR003）