【作者】 李霞；

【导师】 刘瑞芬；

【作者基本信息】 山东中医药大学 ， 中医妇科学， 2008， 博士

【摘要】 目的:从细胞凋亡信号转导的角度探讨化瘀、益气养血、清热中药宫清颗粒防治药物流产后异常子宫出血（简称药流后出血）的作用机制,并探讨流产药物抗早孕的机理,丰富中医学“产后多虚多瘀”理论的科学内涵,为临床防治本病提供实验室生物学依据。方法:研究分现代中医文献研究和临床研究两大部分。现代中医文献研究中,以防治药流后出血的现代中医文献为研究对象,建立证治方药数据库,运用文献研究和统计学分析相结合的方法,从频数分析、因子分析、聚类分析角度对本病病因病机及证治规律进行统计分析。临床研究中,将180例患者随机分为宫清药流组、茜芷药流组及单纯药流组,并设人工流产组（简称人流组）患者60例,在观察临床疗效的同时,收集各组人早孕绒毛、蜕膜组织,透射电镜下观察绒毛及蜕膜组织细胞超微结构变化,并检测细胞凋亡率（AR）、凋亡信号转导相关指标凋亡因子Fas信使核糖核酸（FasmRNA）、Fas配体信使核糖核酸（FasLmRNA）、凋亡因子Fas、Fas配体（FasL）、蛋白激酶C（PKC）、细胞内钙离子浓度([Ca²⁺]i)表达变化,分析指标间的相关性。结果:现代中医文献研究显示,瘀、虚、热是药流后异常子宫出血的基本病因病机,活血化瘀、益气养血、清热是防治本病的基本原则,分析出了防治本病的高频药物及其配伍规律,为以活血化瘀、益气养血、清热法为组方原则的宫清颗粒防治本病提供了中医文献学依据。临床研究发现:宫清药流组孕囊排出时间较茜芷药流组及单纯药流组明显缩短,均有非常显著性差异（P＜0.01）;宫清药流组子宫出血量明显减少,与茜芷药流组比较有显著性差异（P＜0.05）,与单纯药流组比较有非常显著性差异（P＜0.01）;宫清药流组子宫出血天数较单纯药流组明显缩短,有显著性差异（P＜0.05）,与茜芷药流组比较无显著性差异（P＞0.05）。单纯药流组人早孕绒毛及蜕膜组织细胞超微结构观察发现细胞凋亡倾向,凋亡信号转导指标检测显示FasmRNA及FasLmRNA基因表达、Fas及FasL蛋白表达、[Ca²⁺]i明显升高,PKC蛋白表达明显降低,与人流组比较有非常显著性差异（P＜0.01）。宫清颗粒的作用趋势与单纯药流药物相同,其作用效果较茜芷药流组与单纯药流组显著,超微结构观察发现宫清药流组绒毛及蜕膜组织细胞出现典型的凋亡形态学改变,凋亡信号转导指标检测显示,AR、FasmRNA基因及FasLmRNA基因表达、Fas蛋白及FasL蛋白表达、[Ca²⁺]i较茜芷药流组、单纯药流组明显升高（P＜0.05,P＜0.01）,PKC蛋白表达显著降低（P＜0.05,P＜0.01）。宫清药流组AR与FasmRNA基因、FasLmRNA基因、Fas蛋白、FasL蛋白表达及[Ca²⁺]i呈正相关,与PKC蛋白表达呈负相关;FasmRNA基因表达与Fas蛋白表达呈正相关,FasLmRNA基因表达与FasL蛋白表达呈正相关,PKC蛋白表达与[Ca²⁺]i呈负相关。结论:药流药物通过影响Fas/FasL、PKC、Ca²⁺介导的细胞凋亡信号转导途径,引起人早孕绒毛滋养细胞及蜕膜细胞凋亡,这可能是其抗早孕的机理之一。细胞凋亡不仅是药流的启动因素,同时凋亡障碍也是导致药流后异常子宫出血的原因之一。瘀、虚、热是药流后异常子宫出血的基本病因病机,以活血化瘀、益气养血、清热为组方原则的宫清颗粒从基因及蛋白水平调控Fas/FasL、PKC、Ca²⁺介导的细胞凋亡信号转导途径,诱导药流人早孕绒毛滋养细胞及蜕膜细胞凋亡,促进绒毛与蜕膜组织的完全排出,进而达到防治药流后异常子宫出血的目的,其临床疗效及对细胞凋亡信号转导相关指标的调节作用明显优于茜芷药流组和单纯药流组。更多还原

【Abstract】 Objective:To investigate the inducement mechanism of Gongqing granum concerning singnal transduction of human villi and decidua cell apoptosis,which is in the early pregnancy bleeding after drug abortion.In order to provide objective basis for preventing and treating the disease by Gongqing granum,furthermore,enrich the scientific connotation of the TCM theory that "there are more fragility and blood stasis after production".Methods:In the modern TCM literature study,a database of recipes and herbs was founded with computer technology.Then analyzed the therapia rule from the point of frequency analysis,factor analysis and varable clustering analysis.In the clinical experimental investigation,the selected 240 patients were divided into 4 groups by random.They were group of Gongqing,group of Qianzhi,group of drug abortiona and group of induced abortion respectively.We observed the clinical curative effect of the 4 groups,at the same time,we collected the specimen of the 4 groups.Then we observed the ultrastructure of the specimen and detected the expression of apoptosis rate（AR） and correlative indexs of singnal transduction of cell apoptosis,such as FasmRNA,FasLmRNA, Fas,FasL,PKC and intracellular calcium concentration([Ca²⁺])i,furthermore,we analyzed the relativity of the indexs.Results:The TCM literature study showed that the main pathogen of bleeding after drug abortion were blood stasis,fragility and heat,which offered the philology evidence of Gongqing granula preventing and treating bleeding after drug abortion.The clinical experimental investigation showed that the clinical curative effect of Gongqing granum was obviously better than other groups（P＜0.01,P＜0.05）.The observation of ultrastructure indicated distinct change of apoptosis.Compared to the other 3 groups,the expression of AR,FasmRNA,FasLmRNA,Fas,FasL and([Ca²⁺])i of Gongqing group were obviously higher than the other 3 groups（P＜0.01,P＜0.05）,while the expression of PKC of Gongqing group was obviously lower than other 3 groups（P＜0.01, P＜0.05）.Results:Mifepristone can affect the singnal transduction of cell apoptosis caused by singnal transduction Fas/FasL,PKC and Ca²⁺.This may be one of the mechanisms of drug abortion.Blood stasis,fragility and heat are the main reasons of bleeding after drug abortion.Gongqing granum can adjust the signal transduction caused by Fas/FasL,PKC and Ca²⁺,consequently induced the cell apoptosis of human villi and deciduas in early pregnancy.By doing so,Gongqing granum can prevent and treat abnormal uterus bleeding after drug abortion.The effect of clinical curative and adjustion of apoptosis singnal transduction of Gongqing granum are obviously better than Qianzhi and onefold drug abortion.更多还原