痔血胶囊肝毒性药物因素和个体因素研究

发布时间：2018-05-11 07:38

本文选题：痔血胶囊 + 肝毒性　；参考：《北京中医药大学》2017年硕士论文

【摘要】：痔血胶囊是一种纯中药复方制剂,由苦参和白鲜皮两味中药组成,功能主治:清热解毒、凉血止血。用于Ⅰ、Ⅱ期内痔及混合痔所致的便血、肛门坠胀或坠痛,大便干燥或秘结等症。该品种被收入2004年《国家基本药物目录》和《国家基本医疗保险和工伤保险药品目录》。但近年来国家药品不良反应中心陆续收到痔血胶囊涉及的药品不良反应报告,特别是2008年报告数量迅速增加。国家药品不良反应中心第十七期《药品不良反应信息通报》报告了痔血胶囊引起肝损害的不良反应情况:截至2008年9月25日,共收到痔血胶囊相关病例报告35例。其中21例(60%)不良反应名称描述为肝功能异常、胆汁淤积型肝炎、药物性肝炎,另有腹痛8例,皮疹4例,过敏样反应1例,头晕、头痛1例。经对报告病例系统分析后,不能排除痔血胶囊与肝损害事件的关联性。为对痔血胶囊的肝毒性进行全面的研究,本实验从药物因素,以及个体因素,对痔血胶囊出现肝毒性的原因进行了研究,其中药物因素从痔血胶囊中出现过单味药肝损伤报道的白鲜皮的产地、生产工艺;个体因素从辨证、辨体、用量、疗程等角度进行研究。第一部分:药物因素实验一、含不同产地白鲜皮痔血胶囊肝毒性的比较性研究目的:观察分别含不同产地白鲜皮痔血胶囊的长期低(4.1 g.kg-1)、中(8.3 g·kg-1)、高(16.5g·kg-1)剂量给药后对大鼠产生的肝毒性反应。方法:Wistar大鼠,随机分为正常组、含五个不同产地白鲜皮痔血胶囊的高中低剂量组,分别灌服蒸馏水、含不同产地白鲜皮痔血胶囊的高中低剂量,连续给药30 d。观察给药期间的一般表现,体重变化等。末次给药后,检测血清肝功能指标;计算脏器指数;按照肝脏病理检验要求,取肝组织,固定,苏木精-伊红染色(HE染色)后光镜下观察。结果:大鼠连续灌胃含不同产地白鲜皮痔血胶囊30 d,给药期间动物活动、排便、毛发等与正常组无明显差异;30 d后,与正常组比较,肝脏血生化结果白蛋白(ALB)、谷氨酰转肽酶(GGT)在个别组有升高(P0.05);病理切片观察显示,分别含五个不同产地白鲜皮痔血胶囊的动物肝脏低、中、高剂量组均可见透明细胞改变及肝窦扩张。结论:分别含五个不同产地白鲜皮痔血胶囊均具有明显的肝毒性。实验二、痔血胶囊不同工艺提取物肝毒性研究目的:通过比较痔血胶囊水提取物和乙醇提取物对大鼠肝毒性的影响,探讨工艺因素对中药制剂安全性的影响。方法:Wistar大鼠80只,雌雄各半,随机分为空白对照组、全方水提组、全方60%醇提组、全方85%醇提组,除正常组灌服蒸馏水外,其余组按剂量4.1 g·kg-1灌服对应药物,连续给药30 d。末次给药后,腹腔注射10%水合氯醛溶剂麻醉,腹主动脉取血,离心取血清,全自动生化仪检测血清天门冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、丙氨酸氨基转移酶(ALT)、总胆红素(TBIL)等血清肝功能指标;摘取肝脏,称取质量,计算脏器指数;取肝脏最大叶同一部位肝组织,10%中性福尔马林固定48 h,脱水,石蜡包埋,切片,HE染色后,在光学显微镜下进行病理检查。结果:痔血胶囊全方乙醇提取物存在明确的肝脏毒性,组织病理学主要表现为肝小叶周边带肝细胞小泡型脂肪变性;并且其肝毒性与提取溶剂中乙醇含量正相关;此外根据大鼠肝脏小灶状炎细胞浸润评分,发现痔血胶囊的抗炎作用,水提组明显优于醇提组,说明痔血胶囊醇提物较水提物药效降低。结论:工艺因素对中药复方制剂的安全性具有重要影响,提示今后新药研发应关注现代工艺对中药安全性的影响,过分强调成分富集存在严重的安全隐患,工艺筛选应采用毒效结合的思路。实验三、痔血胶囊不同工艺提取物化学成分差异分析目的:运用HPLC-MS联用技术,对痔血胶囊不同工艺提取物化学成分差异进行分析。方法:通过HPLC-MS对痔血胶囊水提取物、85%乙醇提取物、60%乙醇提取物进行分析,对三者的主要成分差异进行OPLS-DA分析,并通过2级图谱对主要差异成分进行鉴定。结果:痔血胶囊水提取物、65%醇提取物、85%醇提取物的化学成分具有明显的含量差异;并初步鉴定其中的已知化合物,为进一步筛选痔血胶囊中的肝毒性成分奠定了基础。第二部分:个体因素通过对痔病的中医证型分布,及其体质分布特征与痔血胶囊主治功能进行比较、以及痔血胶囊肝损伤病例报告的回顾,对造成痔血胶囊肝损伤的个体因素进行分析。发现痔血胶囊出现肝损伤的个体因素主要包括用药辨证不准,用药未考虑体质,以及患者超疗程、超剂量服用等。本研究通过对痔血胶囊中白鲜皮的产地、痔血胶囊的醇提工艺对其肝毒性的影响进行大鼠长期毒性实验,基本明确痔血胶囊出现肝损伤的主要因素之一为工艺因素。但是辨证是否准确、是否辨体用药、以及用药是否符合疗程、用量等个体因素,也是痔血胶囊造成肝损伤的因素之一。
[Abstract]:The hemorrhoid blood capsule is a pure Chinese medicine compound preparation, composed of two herbs of Radix Sophorae Flavescentis and white fresh skin. It is used for the treatment of blood stasis caused by internal hemorrhoids and mixed hemorrhoids in stage I, II period of internal hemorrhoids and mixed hemorrhoids, anus falling or falling pain, the dry or secret of stool. The variety was paid in 2004 < National Basic Medicine > and national basic medical insurance. In recent years, the national drug adverse reaction center has received reports of adverse drug reactions involved in hemorrhoids and blood capsules, especially in 2008. The seventeenth phase of the national drug adverse reaction center report on the adverse drug reaction information report on the adverse reactions caused by hemorrhoids and blood capsules. Situation: as of September 25, 2008, a total of 35 cases of hemorrhoids and blood capsule related cases were reported. Among them, 21 cases (60%) were described as abnormal liver function, cholestasis hepatitis, drug hepatitis, 8 cases of abdominal pain, 4 cases of skin rash, 1 cases of allergic reaction, dizziness and 1 headache. After systematic analysis of the reported cases, the hemorrhoids and blood capsules can not be excluded from the hemorrhoids capsules and blood capsules and can not be excluded. In order to study the hepatotoxicity of the hemorrhoid blood capsule, we studied the hepatotoxicity of hemorrhoids and blood capsules. From the drug factors and individual factors, the cause of the liver toxicity of hemorrhoid capsule was studied. Research on syndrome differentiation, differentiation, dosage and course of treatment. Part 1: drug factor experiment 1. Comparative study on hepatotoxicity of hemorrhoid capsules containing white fresh skin in different habitats: long-term low (4.1 g.kg-1), medium (8.3 G. Kg-1), and high (16.5g. Kg-1) dosage of high (16.5g. Kg-1), respectively, of different producing areas Methods: Wistar rats were randomly divided into normal group and high and low dose group containing five different areas of white fresh skin hemorrhoids and blood capsules. The low dose of distilled water and high and low doses of white fresh skin hemorrhoid capsule in different areas were given respectively. The general performance and weight change during the period of 30 D. were continuously given. After the last administration, the liver function was detected. Index; calculate the organ index; take the liver tissue, fixed, the hematoxylin eosin staining (HE staining) under the light microscope according to the liver pathological examination requirements. Results: the rats were continuously filled with 30 d capsules containing white fresh skin hemorrhoids and blood, and there was no significant difference between the animal activities, defecation, hair and so on during the period of administration. After 30 d, the liver blood was compared with the normal group. The biochemical results of albumin (ALB) and glutamyl transpeptidase (GGT) were elevated in a few groups (P0.05). Pathological sections showed that the liver of animals with five different areas of white fresh skin hemorrhoid capsules were low, middle and high dose group showed transparent cell changes and hepatic sinusoid dilatation. Significant liver toxicity. Experiment two, study on the hepatotoxicity of different process extracts of hemorrhoid blood capsules. By comparing the effects of water extract and ethanol extract of hemorrhoid Capsule on the liver toxicity of rats, the effect of technological factors on the safety of Chinese medicine preparation was discussed. Methods: 80 Wistar rats were divided into blank control group and the whole water extract group at random. The whole 60% alcohol extraction group and the whole 85% alcohol extract group, except the normal group was filled with distilled water, the other groups were given the corresponding drugs according to the dosage of 4.1 G. Kg-1. After continuous administration of the last dosage of 30 d., the abdominal aorta was injected with chloral chloral solvent anesthesia, the abdominal aorta was taken from the blood, the serum was centrifuged, and the serum aspartate aminotransferase (AST) was detected by the automatic biochemistry analyzer. Phosphatase (ALP), alanine aminotransferase (ALT), total bilirubin (TBIL) and other serum liver function indexes, extract the liver, take the mass, calculate the organ index; take the liver tissue of the same part of the liver of the largest lobe of the liver, 10% neutral Faure Marin fixed 48 h, dehydration, paraffin embedding, slice, HE staining, pathological examination under the optical microscope. Results: hemorrhoids blood There was a clear liver toxicity in the whole capsule ethanol extract of the capsule. Histopathology was mainly manifested by hepatocyte vesicular steatosis in the periphery of the liver lobule, and the liver toxicity was positively correlated with the ethanol content in the extraction solvent. The results showed that the technological factors had an important influence on the safety of Chinese medicine compound preparation, suggesting that the development of new drugs should pay attention to the effect of modern technology on the safety of traditional Chinese medicine, overemphasize the existence of serious safety hidden dangers in the enrichment of ingredients, and the process screening should be combined with toxic effect. Experiment three, the difference analysis of chemical composition of different process extracts of hemorrhoid blood capsule: use HPLC-MS combined technique to analyze the chemical composition difference of different process extracts of hemorrhoid blood capsule. Method: analysis the water extract of hemorrhoid blood capsule, 85% ethanol extract and 60% ethanol extract through HPLC-MS. OPLS-DA analysis was carried out and the main difference components were identified by 2 level atlas. Results: the chemical composition of the water extract of hemorrhoid blood capsule, 65% alcohol extract and 85% alcohol extract had obvious difference, and the preliminary identification of the known compounds was established. Second parts were laid for the further screening of the hepatotoxic components in hemorrhoid blood capsules. The individual factors were compared with the TCM syndrome distribution of hemorrhoids, the distribution characteristics of the constitution of hemorrhoids and the main function of hemorrhoids and blood capsules, and the review of the report on the case of hemorrhoid capsule liver injury, and the individual factors that caused the liver injury of hemorrhoids and blood capsules were analyzed. In this study, the effect of the alcohol extraction process on the liver toxicity of hemorrhoids and blood capsules was carried out in the long-term toxicity test on the liver toxicity of hemorrhoids and blood capsules. Whether or not body identification is used, and whether the medication is in line with the course of treatment and dosage is also one of the factors that cause liver injury caused by hemorrhoids capsule.

【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285

【参考文献】