基于益智仁盐炙前后及分别组成缩泉丸来研究“盐炙入肾-肾主水”的机制

"Salt-roasted herbs tending to the kidney- the kidney controlling water" is one of the basical theories of Chinese medicinal herbs preparation and basic theories of traditional Chinese medicine.But at present the theory is still explained by"salt tasting salty, salty flavor tending to the kidney" modern scientific content need to be revealed and clarified. In support of projects such as National Natural Science Foundation and The National 11th five-year technology based plan topic,our task group has completed the research of the changes of chemical constituents and effects before and after preparation of Fructus Alpinia Oxyphylla by chemical and pharmacological methods,as well as the research of the scientific connotation of"salt-roasted herbs tending to the kidney-warming the kidney and reducing urine"by analyzing the expression of protein and gene of AQP-2 in the kidney.However, traditional Chinese medicine should be mostly used in the form of prescriptions.So our project plans to put Fructus Alpinia Oxyphylla before and after preparation into the prescription"Suoquanwan"and combine its effect of warming the kidney and reducing urine,in order to explore the processing mechanism of "salt-roasted herbs tending to the kidney".Furthermore,warming the kidney and reducing urine have significantly related to the TCM theory of "the kidney controlling water".The research model of "spectrum-activity relationship→ the isolation and identification of effective components→ dynamic changes of effective components in vivo"will be applied to perfect the explaination of the processing mechanism of "salt-roasted herbs tending to the kidney"of Fructus Alpinia Oxyphylla. Adopting serum fingerprint and PK-PD model can help to clarify the change law of chemical constitutions in Suoquanwan before and after preparation of Fructus Alpinia Oxyphylla.Advanced techniques such as cell culture,real-time quantitative and western blot will be used to clarify the scientific connotation of "salt-roasted herbs tending to the kidney- the kidney controlling water"and then to propel the development of science of Chinese medicinal herbs preparation.

"盐炙入肾-肾主水"是中药炮制及中医基本理论之一，目前仍以"盐味咸，咸入肾"的传统理论解释，缺乏现代科学内涵的阐明。课题组前期在国家自然科学基金和"十一五"支撑计划等项目支持下，对单味益智仁经化学、药理等研究，揭示其盐炙前后化学成分及药效改变，并从肾脏AQP-2基因和蛋白表达层面初步阐明其"盐炙入肾-温肾缩尿"的科学内涵。但中医用药以复方为主，故本课题将益智仁盐炙前后分别组成"缩泉丸"，并结合"缩泉丸"温肾缩尿功效来研究其"盐炙入肾"炮制机制，而温肾缩尿与中医"肾主水"的基本理论密切相关。拟采用"谱效关系→药效成分分离鉴定→药效成分体内动态变化"研究模式完善益智仁"盐炙入肾"炮制机制；进一步通过"血清指纹图谱→药效成分PK-PD"研究模式揭示益智仁盐炙前后在复方中的变化规律；并引入细胞培养，实时荧光定量，蛋白印迹法等先进技术，以阐明益智仁"盐炙入肾-肾主水"的科学内涵，促进中药炮制学科发展

缩泉丸出自《妇人良方》，由盐益智仁、乌药、山药组成。方中君药益智仁炮制品的选择是影响其临床疗效的关键，因益智仁生品主入脾经，盐炙后主入肾经，主治肾虚小便清长，夜尿频多或遗尿。在前期研究的基础上，课题组多层次阐明了益智仁“生品入脾·止泻，盐炙入肾·缩尿”中药炮制理论的科学内涵。本项目在前期研究的基础上，以缩泉丸“补肾缩尿”的功效为切入点，通过谱效关系筛选药效成分，再分离鉴定，通过吸收、分布、代谢、排泄研究成分体内动态变化；并通过比较益智仁炮制前后及纳入复方缩泉丸中血清指纹图谱及主要药效成分药代动力学差异，揭示缩泉丸中益智仁盐炙前后在体内的变化规律；通过益智仁盐炙前后组成的缩泉丸对不同模型大鼠肾脏、膀胱影响尿液生成和排放受体等的影响，多层次多角度阐明"盐炙入肾-肾主水"的科学内涵。结果如下：.(1)通过灰关联分析-成分分离-药效学分析，结果表明盐炙益智仁石油醚部位分离得到的7-epi-teucrenone B 可能是通过拮抗磷酸组胺的H1 受体释放从而抑制豚鼠膀胱逼尿肌收缩张力。.(2)通过比较益智仁盐炙前后纳入缩泉丸指纹图谱比较，说明缩泉丸中益智仁盐炙后化学成分有所改变。通过比较缩泉丸中益智仁盐炙前后的血清指纹图谱，发现二者在入血成分上差别不大，但各共有峰平均峰面积有所改变，提示益智仁盐炙后组成缩泉丸疗效增强可能与入血成分的含量上的变化有关。而药动学结果显示益智仁盐炙后组成的缩泉丸不仅提高了君药益智仁入血成分生物利用度，同时还提高了配伍药物的生物利用度。.(3)益智仁盐炙后组成的缩泉丸其缩尿作用增强，是通过改善肾脏和睾丸病理损伤，改善肾功能，提高免疫功能，三个方面增强药物“补肾”作用的，从三个方面阐明了缩泉丸中益智仁“盐炙入肾”的作用机理。.(4)益智仁盐炙后组成的缩泉丸增强水液代谢平衡是通过提高肾脏AQP-2的表达增加对水的重吸收，调节M3R 和 β3-AR的表达维持膀胱正常收缩和舒张的功能，调节神经内分泌功能三个方面进行的，阐明了缩泉丸中益智仁盐炙“肾主水”的作用机理。