痔血胶囊水提物和醇提物对大鼠肝毒性的影响

目的:通过比较痔血胶囊水提物和醇提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%水合氯醛麻醉大鼠,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%中性甲醛固定48 h,脱水,石蜡包埋,切片,苏木素-伊红(HE)染色后,在光学显微镜下进行病理检查。结果:痔血胶囊全方醇提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。

Effect of Water Extract and Ethanol Extract of Zhixue Capsule on Hepatotoxicity in Rats

Objective: To explore the effect of process factors on the safety of traditional Chinese medicine(TCM) preparation by comparing the effect of water extract and ethanol extract of Zhixue capsule on hepatotoxicity. Method: The 80 Wistar rats (40 male and 40 female) were randomly divided into blank control group, Zhixue capsule water extract group, Zhixue capsule 60% ethanol extract group, and 85% ethanol extract group. The rats in normal group received distilled water by gavage, and the rats in other groups received 4.1 g·kg^-1 extraction by gavage for 30 days. Intraperitoneal injection of 10% chloral hydrate was used to anesthetize the rats after the last drug administration, then blood was taken from abdominal aorta and serum was taken by centrifugation. Aspartate aminotransferase (AST),alkaline phosphatase (ALP), alanine aminotransferase (ALT), and total bilirubin (TBIL) in serum were detected to indicate the liver injury. Their livers were taken for weighing and calculating the viscera index, the liver tissues in the same place of the largest liver lobe were taken and fixed by 10% neutral formalin for 48 h, then dehydration, paraffin embedding, and slicing were conducted. After hematorylin-eosin(HE) staining, pathological examination was done under the microscope. Result: Zhixue capsule ethanol extract had clear hepatotoxicity, histopathologically demonstrated as small bubble type fatty degeneration surrounding hepatic lobule, and its hepatotoxicity was positively correlated with the ethanol concentration. In addition, water extract group had significantly better anti-inflammatory effect than ethanol extract group according to the liver focal necrosis and inflammatory cells infiltration scores, indicating that the effect of Zhixue capsule ethanol extract was lower than that of water extract. Conclusion: Processing factors have important effects on the safety of TCM compound preparations, indicating that the impact of modern technology on safety shall be concerned in new drug research and development, serious safety risks shall be emphasized in composition enrichment, and toxicity-effect thinking shall be adopted in technology screening.