目的 对主产地广东遂溪、福建漳浦及引种栽培地区江苏南京、淮安的不同生长期穿心莲Andrographis paniculata活性成分及关键酶基因表达情况进行分析,研究穿心莲活性成分积累动态,探讨其最佳采收期及分子机制。方法 用HPLC法测定穿心莲内酯及脱水穿心莲内酯量,并用实时荧光定量PCR技术从mRNA水平比较研究不同生长期穿心莲关键酶ent-柯巴基焦磷酸合酶(CPS)基因的表达情况。结果 不同产地穿心莲生长过程基本一致,在营养生长时期生长迅速,进入生殖生长后生长趋于平缓。4个产地穿心莲样品中活性成分变化趋势相近,穿心莲内酯量均在始花期达到最高,脱水穿心莲内酯量变化较平缓;穿心莲CPS基因的相对表达量在蕾期至始花期相对较高,与穿心莲内酯量呈显著正相关。4个产地穿心莲的穿心莲内酯与脱水穿心莲内酯之和均达到《中国药典》2010年版标准。结论 穿心莲最佳采收期应以物候期为评判依据,其最佳采收期为始花期。

Objective To study the dynamic changes of the active components from Andrographis paniculata at various growth periods grown in Suixi in Guangdong province, Zhangpu in Fujian province as well as introduced and cultivated in Nanjing and Huaian in Jiangsu province, and to investigate the expression level of the key enzyme gene at the same time in order to obtain the accumulation trend of the active contents from A. paniculata and explore a reasonable harvest and its molecular mechanism. Methods The contents of andrographolide and dehydroandrographolide were analyzed by HPLC. The expression of key enzyme gene, ent-copalyl diphosphate synthase (CPS) was measured at mRNA level with QRT-PCR. Results The growth periods of A. paniculata were similar in different areas. It grew fast at vegetative stage while slowly at reproductive stage. The contents of active ingredients in A. paniculata from four areas showed the same change tendency. The content of andrographolide reached the highest level at flowering period. At the same time, the changes of dehydroandrographolide was flatting out. The expression of CPS reached its maximum at bud to flowering period. There was a significant positive correlation between the content of andrographolide and the expression of CPS. Conclusion The optimal harvest time of A. paniculata should be judged on phenological period and the best harvest period is flowering period.

国家"十二五"科技支撑项目(2012BAI29B02);江苏省青蓝工程(2012);江苏省中药优势学科II期建设(ysxk-2014);国家自然基金重点项目中药道地性及其形成的环境机制研究(81130070);道地药材国家重点实验室培育基地建设项目