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| 五灵胶囊工艺优化前后对四氯化碳诱导的肝纤维化大鼠保护作用的比较 |
| 陈琳1,曹凡2,宋忠兴1,孔鑫2,马梦鸽3,唐志书1，4*,刘红娜5,周建平5,曹兆军5 |
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(1. 陕西省中药资源产业化协同创新中心, 秦药特色资源研究开发国家重点实验室(培育), 陕西省创新药物研究中心, 陕西中医药大学, 咸阳 712083;
2. 陕西中医药大学药学院, 西安 712046;
3. 长春中医药大学药学院, 长春 130117;
4. 中国中医科学院研究生院, 北京 100700;
5. 清华德人西安幸福制药有限公司, 西安 710043
*通信作者) |
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| 摘要: |
| 目的 比较五灵胶囊工艺优化前后对四氯化碳（CCl4）诱导的肝纤维化大鼠的保护作用。方法 将SD大鼠随机分为9组：对照组，模型组，水飞蓟素阳性药组，五灵胶囊新工艺低（0.25 g/kg）、中（0.5 g/kg）、高（1.0 g/kg）剂量组，五灵胶囊原工艺低（0.25 g/kg）、中（0.5 g/kg）、高（1.0 g/kg）剂量组，每组10只。除对照组外，其余各组大鼠腹腔注射40% CCl4（橄榄油配制，剂量为2 mL/kg），每周2次，连续8周，构建肝纤维化模型。给药组于第2周开始灌胃给药，持续6周。实验结束后称取大鼠肝脏、脾脏湿重并计算脏器指数；检测血清生化指标丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、碱性磷酸酶（ALP）、总胆汁酸（TBA）、白蛋白（ALB）含量和超氧化物歧化酶（SOD）活性；采用H-E染色和Masson染色进行肝组织病理结构和纤维化程度观察；应用免疫组织化学染色检测肝组织中α-平滑肌肌动蛋白（α-SMA）水平；采用ELISA测定血清中炎症因子IL-6、IL-1β和TNF-α含量；采用qPCR法测定肝脏中胶原蛋白（COL）Ⅰ、COL Ⅲ、COL Ⅳ和NF-κB mRNA的相对表达量。结果 五灵胶囊优化工艺前后均改善了模型大鼠的肝脏病理结构、降低了肝纤维化程度。与模型组相比，五灵胶囊新工艺和原工艺组大鼠血清中ALT、AST、ALP和TBA含量均降低（P＜0.05）；血清中ALB含量和SOD活性均升高（P＜0.05）；肝脏组织中α-SMA阳性表达面积均减少（P＜0.05）；血清中IL-6、IL-1β和TNF-α含量均降低（P＜0.05）；肝脏组织中COL I、COL Ⅲ、COL Ⅳ和NF-κB的mRNA表达水平均下调（P＜0.05），其中五灵胶囊新工艺组TBA、α-SMA和COL Ⅲ优于原工艺组。结论 五灵胶囊新工艺和原工艺对CCl4诱导的大鼠肝纤维化均具有保护作用，但新工艺的部分指标优于原工艺。 |
| 关键词: 五灵胶囊 工艺优化 四氯化碳 肝纤维化 抗炎 |
| DOI：10.16781/j.CN31-2187/R.20211157 |
| 投稿时间：2021-11-16修订日期：2022-01-06 |
| 基金项目:国家现代农业产业技术体系资助项目（CARS-21），咸阳市2020年度重大科技专项（2020K01-20），中药大品种品牌价值提升示范研究项目（202190025），陕西省千人计划区域青年人才项目（2018-11），陕西省教育厅重点科学研究计划项目（22JY019）. |
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| Protective effects of Wuling capsule on CCl4-induced hepatic fibrosis in rats before and after process optimization |
| CHEN Lin1,CAO Fan2,SONG Zhong-xing1,KONG Xin2,MA Meng-ge3,TANG Zhi-shu1，4*,LIU Hong-na5,ZHOU Jian-ping5,CAO Zhao-jun5 |
(1. Shaanxi Collaborative Innovation Center of Chinese Medicine Resources Industrialization, State Key Laboratory of Research and Development of Characteristic Qin Medicine Resources(Cultivation), Shaanxi Innovative Drug Research Center, Shaanxi University of Chinese Medicine, Xianyang 712083, Shaanxi, China;
2. College of Pharmacy, Shaanxi University of Chinese Medicine, Xi'an 712046, Shaanxi, China;
3. College of Pharmacy, Changchun University of Chinese Medicine, Changchun 130117, Jilin, China;
4. Graduate School, China Academy of Traditional Chinese Medicine, Beijing 100700, China;
5. Tsinghua Deren Xi'an Happiness Pharmaceutical Co., Ltd., Xi'an 710043, Shaanxi, China
*Corresponding author) |
| Abstract: |
| Objective To compare the protective effects of Wuling capsule on CCl4-induced hepatic fibrosis rats before and after process optimization. Methods SD rats were randomly divided into 9 groups: control group, model group, positive control group (silymarin) and low (0.25 g/kg)-, medium (0.5 g/kg)- and high (1.0 g/kg)-dose of Wuling capsule new process and original process groups, with 10 rats in each group. The rat model with liver fibrosis was established by intraperitoneal injection of CCl4 solution (dissolved in olive oil, at a dosage of 2 mL/kg), twice a week for 8 weeks except control group. The rats in the treatment groups were administered by gavage at the second week for 6 consecutive weeks. At the end of the experiment, the wet weights of the rat liver and spleen were examined and the viscera indexes were calculated. Serum biochemical indexes (alanine aminotransferase ［ALT］, aspartate aminotransferase ［AST］, alkaline phosphatase ［ALP］, total bile acid ［TBA］, and albumin ［ALB］) and superoxide dismutase (SOD) activity were detected. Hematolyxin-eosin staining and Masson staining were used to observe the pathological structure and fibrosis degree of liver tissues. The level of α-smooth muscle actin (α-SMA) in rat liver tissues was detected by immunohistochemistry. The levels of interleukin (IL)-6, IL-1β and tumor necrosis factor α (TNF-α) in serum were determined by enzyme-linked immunosorbent assay. The mRNA relative expression levels of collagen (COL) Ⅰ, COL Ⅲ, COL Ⅳ and nuclear factor κB (NF-κB) in the liver were determined by quantitative polymerase chain reaction (qPCR). Results Both the new and original process groups of Wuling capsule improved the liver pathological structure of CCL4-induced hepatic fibrosis rats and reduced the degree of hepatic fibrosis. Compared with the model group, the contents of ALT, AST, ALP and TBA in serum were significantly decreased (P＜0.05); serum ALB content and SOD activity were significantly increased (P＜0.05); the α-SMA positive area ratio in liver tissues decreased (P＜0.05); the contents of serum IL-6, IL-1β and TNF-α were significantly decreased (P＜0.05); and the mRNA expression levels of COL Ⅰ, COL Ⅲ, COL Ⅳ and NF-κB in liver tissues were significantly down-regulated (P＜0.05). The experimental results of TBA, α-SMA and COL Ⅲ in the new process group were significantly better than those in the original process group. Conclusion Both the new technology and original technology of Wuling capsule have protective effects on CCl4-induced hepatic fibrosis in rats, but some indexes of the new technology are better than those of the original technology group. |
| Key words: Wuling capsule process optimization carbon tetrachloride liver fibrosis anti-inflammation |
