铁霜替朱砂消除朱砂安神丸毒性的"量-效-毒"关系原理基础研究

The study is aimed to replace the heavily toxic component Cinnabar (CNB) in Zhushaanshen Pill (ZSASP) by Ferrous acetate (FAC), for they are both important sedative medicines, but FAC with much lower toxicity which is useful for hemoglobin production.Firstly the toxicity, equal efficacy dosage and metabolismics test of CNB and FAC will be systematically studied,in which the best compatibility proportionality of Tieshuanganshen Pill (TSASP) will be prescribed through orthogonal design combined with the sedative efficacy experiment for rats after administration and the results will serve as the assessing criteria. Meanwhile, the characteristic fingerprints associated with active efficacy for TSASP will be established to find a group of effective components to be served as the quantitative control targets also combined with the quantified fingerprints to control TSASP quality effectively. Secondly,the systematic pharmacodynamic tests will be performed,such as the subacute and subchronic toxic dose tests are employed to investigated how about the nephrotoxity of TSASP and ZSASP.Through using metabonomics method, we will investigate the variances of metabolites in blood, urine, and kidney caused by them. After then we can find the equal potency dosage in clinics for TSASP instead of ZSASP. Moreover, thoroughly eliminating the renal toxicity of ZSASP.Thirdly, Study the hypnotic mechanism of replacement formulation at the molecular level,for 48 hours of sleep deprivation treatment group and the control group① to detect how about the rat hippocampus, cortex and cerebellum c-fos, nestin, bid3 and jak2 expression levels;②to detect how about rat hippocampus, cortex and cerebellum Fos, Nestin, Bid3 and Jak2 protein levels detected.The final objective grealy focus on relationship theory of quantity–efficacy-toxicity for TSASP, which provide a typical model for natural compound TCM with more effective, non-toxic and stable quality control for all over the world patients.The important fundmental sciencitific study for FAC instead of CNB will thoroughly cancel the renal toxicity of CNB, which provide a novel model for the more than 60 kinds of Compound TCM in Ch.P.taking TSASP as an example.

对铁霜替朱砂安神丸毒药味-朱砂开展研究。铁霜和朱砂都是重镇安神药，但铁霜毒性很低，为醋酸亚铁，利于血红蛋白生成。第一，对朱砂、铁霜的毒性、等效药量和代谢组学研究。通过小鼠镇静试验确定铁霜替朱砂的最佳配伍，建立特征谱效学,找到镇静安神组效物质作质量控制指标，并采用定量指纹图谱控制。第二，经药效试验、亚急和亚慢肾毒性试验研究铁霜安神丸和朱砂安神丸肾毒性；用代谢组学监测肾毒性及损伤状况，建立铁霜安神丸临床生物等效量。第三，在分子水平上研究替换制剂催眠机制：睡眠剥夺48小时给药组和对照组①检测大鼠海马、皮质和小脑中c-fos,nestin,bid3和jak2表达水平；②检测大鼠海马、皮质和小脑中Fos,Nestin,Bid3和Jak2蛋白表达水平。目标是要消除朱砂安神丸肾毒性，建立铁霜替朱砂的量-效-毒基础关系原理，找到疗效好、低毒、质量可控的催眠药,对中国药典收录的60多个含朱砂方提供替代新方法

本课题通过考察朱砂和铁霜对小鼠自主活动及阈下剂量和阈剂量戊巴比妥钠镇静催眠作用的影响，确定二者等效剂量，明确了朱砂与铁霜二者替换的可行性。采用正交设计得到铁霜安神方的配伍方案，并建立了双波长串联HPLC指纹图谱，基于Sm、Pm和α，用系统指纹定量法（SQFM）对不同组方配比铁霜安神方质量进行了量化评价，通过药效学检测得到了各组铁霜安神方的镇静催眠作用结果，结合灰色度关联分析和多元线性回归分析构建了谱效关系，阐明了铁霜安神方特征指纹图谱所代表的化学成分对镇静催眠药效作用贡献率的大小，同时通过与朱砂安神丸镇静催眠药效的比较研究，实现了对铁霜安神方最佳配伍方案的优化；通过对给予铁霜安神方治疗的失眠大鼠的肝肾组织苏木精-伊红（hematoxylin-eosin，HE）染色切片的观察，进一步确立了铁霜替朱砂消除朱砂安神丸毒性用药的安全性和可行性；采用撤药分析法，验证了铁霜药味对铁霜安神方镇静催眠作用的贡献及重要作用。综上，课题建立了铁霜替朱砂消除朱砂安神丸毒性的“量-效-毒”关系原理基础，找到了疗效好、低毒、质量可控的镇静催眠药，并对中国药典收录的70个含朱砂方提供替代新方法。对亚铁离子和二价汞离子与多巴胺络合催眠机理进行了研究，以及对相关催眠基因组学和蛋白组学的研究。