利用生物效应比评价利咽丹质量的方法研究

目的:测定利咽丹(Liyan Dan,LYD)及其药效组分(active components alignment,ACA)抗Ⅰ型单纯疱疹病毒(HSV-Ⅰ)的生物效应比,探讨使用药效组分标准生物效价表述中药质量的新鉴定方法。方法:用MTT染色法检测对HSV-Ⅰ所致细胞病变(cytopathogenoc effect,CPE)的抑制作用,用HPLC和UV法检测药效组分。用利咽丹及其药效组分对感染HSV-Ⅰ的细胞保护率(protective percent,PP)计算其生物效应比(biologlical effect ratio,BER)。BERS=PPS／PPACA(药物对细胞的保护率／药效组分对细胞的保护率;药物浓度2-x mg/mL)。结果:利咽丹有抑制HSV-Ⅰ所致CPE的活性,其生物效应比为1．16。结论:中药及其药效组分的生物效应比可以反映和揭示其品质的内涵,该指标可以客观地表述中药疗效的本质。本研究提出了中药品质研究的新思路和方法,引入了生物效应比和药效组分的新概念。中药药效组分生物效应质量鉴定方法如下:第一,建立药效组分标准生物效应作为效应标准;第二,按照药效组分生物效应标准的检测方法测定供试品;第三,按照公式计算生物效应比(BERS=PPS／PPACA)。中药药效组分质量评价生物效应比标准:1≥BERS≥0．9时,为合格药品;BERS>1时为优质药品;BERS<0．9时为不合格药品。此质量评价方法比仅依靠一种或两种化学成分含量来评价质量要更科学。

Research of Quality Evaluation Method of Liyan Dan Complex Pharmaceutics by Biological Effect Ratio

Objective: To determine biological effect ratio (BER) on anti-herpes simplex virus-Ⅰ (HSV-Ⅰ)of Liyan Dan (LYD) complex pharmaceutics and its active components alignment (ACA), and to discuss the new identification method of Chinese patent medicine expressed by standard BER. Methods: Protective effect to cells infected by HSV-Ⅰ was evaluated by a 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The ACA was determined by high performance liquid chromatography (HPLC) and ultraviolet spectrophotometry. By the protective percent (PP) to cells infected by HSV-Ⅰ of LYD and its ACA the BER was calculated. BERS= PPS / PPACA (protective percent to cells infected by HSV-Ⅰ of sample/protective percent to cells infected by HSV-Ⅰ of ACA; the concentration of sample was 2-x mg/mL). Results: LYD can inhibit cytopathogenic effect (CPE) and its BER was 1.16. Conclusion: BER and ACA of Chinese patent medicine can reflect connotation of Chinese patent medicine quality. This index described intrinsic quality of curative effect on Chinese materia medica.The new concepts of BER and ACA were pulled into the study. This research offered a new train of thought and method to study Chinese patent medicine quality as follows. Firstly, establish standard biological effect of ACA as a standard of effect; secondly,measure the biological effect of sample by the method of measuring effective standard of ACA; thirdly, calculate BER according to the formula (BERS=PPS/PPACA). Standard of BER on quality evaluation is as follows. If 1 ≥ BERS ≥ 0.9, the sample is eligible; if BERS＞1, the sample is of high quality; if BERS＜0.9, the sample is disqualification. This quality evaluation method is much more scientific than the method that evaluate quality only by one or two components contents.