中国中西医结合外科杂志
中國中西醫結閤外科雜誌
중국중서의결합외과잡지
CHINESE JOURNAL OF SURGERY OF INTEGRATED TRADITIONAL AND WESTERN MEDICINE
2015年
4期
379-382
,共4页
浙贝母%P-糖蛋白%肿瘤耐药%活性组分
浙貝母%P-糖蛋白%腫瘤耐藥%活性組分
절패모%P-당단백%종류내약%활성조분
Bulbus Fritillariae Thunbergii%P-glycoprotein%tumor multidrug resistance%active component
目的：阐明浙贝母抑制耐药肿瘤细胞P-糖蛋白（P-gp）的活性组分。方法：利用回流提取法通过树脂纯化技术，制备浙贝母总生物碱、总核苷、总多糖。建立HepG2/mdr细胞株。利用Calcein-AM试剂盒HepG2/mdr细胞摄取实验对浙贝母总生物碱、总核苷、总多糖进行P-gp潜在抑制剂的筛选。结果：制备的浙贝母总生物碱、总核苷、总多糖含量为51.45%、47.84%、23.78%。HepG2/mdr细胞株P-gp的相对蛋白表达为(26.22±1.09)，极显著高于亲本HepG2细胞株(8.91±2.00)（P＜0.01）。阳性对照组Calcein荧光比率为(173.96±12.87)%，极显著高于空白组(100.00±11.49)%（P＜0.01）。浙贝母总生物碱、总核苷低剂量组Calcein荧光比率为(134.96±11.91)%、(126.89±10.42)%，显著高于空白组(100.00±11.49)%（P＜0.05）。浙贝母总生物碱、总核苷高剂量组Calcein荧光比率为(209.72±9.44)%、(162.42±7.83)%，极显著高于空白组(100.00±11.49)%（P＜0.01）。浙贝母总多糖低、高剂量组与空白组无显著差异。结论：HepG2/mdr细胞株表现出耐药性，阳性抑制剂抑制P-gp的外排活性；浙贝母总生物碱、总核苷抑制P-gp的外排活性，呈现浓度依赖性。
目的：闡明浙貝母抑製耐藥腫瘤細胞P-糖蛋白（P-gp）的活性組分。方法：利用迴流提取法通過樹脂純化技術，製備浙貝母總生物堿、總覈苷、總多糖。建立HepG2/mdr細胞株。利用Calcein-AM試劑盒HepG2/mdr細胞攝取實驗對浙貝母總生物堿、總覈苷、總多糖進行P-gp潛在抑製劑的篩選。結果：製備的浙貝母總生物堿、總覈苷、總多糖含量為51.45%、47.84%、23.78%。HepG2/mdr細胞株P-gp的相對蛋白錶達為(26.22±1.09)，極顯著高于親本HepG2細胞株(8.91±2.00)（P＜0.01）。暘性對照組Calcein熒光比率為(173.96±12.87)%，極顯著高于空白組(100.00±11.49)%（P＜0.01）。浙貝母總生物堿、總覈苷低劑量組Calcein熒光比率為(134.96±11.91)%、(126.89±10.42)%，顯著高于空白組(100.00±11.49)%（P＜0.05）。浙貝母總生物堿、總覈苷高劑量組Calcein熒光比率為(209.72±9.44)%、(162.42±7.83)%，極顯著高于空白組(100.00±11.49)%（P＜0.01）。浙貝母總多糖低、高劑量組與空白組無顯著差異。結論：HepG2/mdr細胞株錶現齣耐藥性，暘性抑製劑抑製P-gp的外排活性；浙貝母總生物堿、總覈苷抑製P-gp的外排活性，呈現濃度依賴性。
목적：천명절패모억제내약종류세포P-당단백（P-gp）적활성조분。방법：이용회류제취법통과수지순화기술，제비절패모총생물감、총핵감、총다당。건립HepG2/mdr세포주。이용Calcein-AM시제합HepG2/mdr세포섭취실험대절패모총생물감、총핵감、총다당진행P-gp잠재억제제적사선。결과：제비적절패모총생물감、총핵감、총다당함량위51.45%、47.84%、23.78%。HepG2/mdr세포주P-gp적상대단백표체위(26.22±1.09)，겁현저고우친본HepG2세포주(8.91±2.00)（P＜0.01）。양성대조조Calcein형광비솔위(173.96±12.87)%，겁현저고우공백조(100.00±11.49)%（P＜0.01）。절패모총생물감、총핵감저제량조Calcein형광비솔위(134.96±11.91)%、(126.89±10.42)%，현저고우공백조(100.00±11.49)%（P＜0.05）。절패모총생물감、총핵감고제량조Calcein형광비솔위(209.72±9.44)%、(162.42±7.83)%，겁현저고우공백조(100.00±11.49)%（P＜0.01）。절패모총다당저、고제량조여공백조무현저차이。결론：HepG2/mdr세포주표현출내약성，양성억제제억제P-gp적외배활성；절패모총생물감、총핵감억제P-gp적외배활성，정현농도의뢰성。
Objective To clarify P-glycoprotein (P-gp) inhibitor of multidrug resistant tumor in Bulbus Fritillariae Thunbergii. Methods Alkaloid, nucleoside and polysaccharide from Bulbus Fritillariae Thunbergii were got by reflux method and resin purification technology. HepG2/mdr cell line was established. The potential P-gp inhibitor on alkaloid, nucleoside and polysaccharide from Bulbus Fritillariae Thunbergii was screened by cellular uptake using the Calcein-AM Kit. Results The contents of preparative alkaloid, nucleoside and poly?saccharide from Bulbus Fritillariae Thunbergii were 51.45%, 47.84% and 23.78% alternatively. The relative pro?tein express of HepG2/mdr was (26.22 ± 1.09), very significantly higher than that of HepG2 (8.91 ± 2.00) (P<0.01). The Calcein fluorescence percentage of positive group was (173.96±12.87)%, very significantly higher than that of the control group (100.00 ± 11.49)% (P<0.01). The Calcein fluorescence percentages of low-dose alka?loid group and nucleoside group were (134.96±11.91)% and (126.89±10.42)%, significantly higher than those of the control group (100.00 ± 11.49)% (P<0.05). The Calcein fluorescence percentages of high-dose alkaloid group and nucleoside group were (209.72 ± 9.44)% and (162.42 ± 7.83)%, very significantly higher than those of the control group (100.00±11.49)% (P<0.01). The Calcein fluorescence percentage of polysaccharide group had no significant differences from that of the control group. Conclusion Established HepG2/mdr shows multi?drug resistance; the positive inhibitor inhibits P-gp efflux activity; alkaloid and nucleoside from Bulbus Fritillari?ae Thunbergii inhibit P-gp efflux activity and show concentration dependence.
