摘要
目的:研究扶正通络法验方芪甲柔肝方抗肝纤维化的疗效与机制。方法:采用皮下注射四氯化碳(CCl4)花生油+10%乙醇代替饮水的方式建立大鼠肝纤维化模型,随机将大鼠分为空白组,模型组,芪甲柔肝方高、中、低剂量组及鳖甲煎丸组,分别以0.9%氯化钠溶液,高、中、低剂量芪甲柔肝方水煎剂,鳖甲煎丸混悬液灌胃,疗程为6周。检测肝功能、肝指数与血清纤维化标志物以判断疗效,并用qRT-PCR检测大鼠肝miR-23a-3p的表达,结合生物信息学分析其调控机制。结果:(1)与空白组比较,模型组大鼠ALT、AST、肝指数及血清纤维化标志物(PⅢP、PCⅣ、HA、LN)显著升高(P<0.05),肝组织显示明显的炎症与纤维化,并出现大量假小叶。与模型组比较,芪甲柔肝方高、中、低剂量组及鳖甲煎丸组均能在一定程度上改善上述指标,其中芪甲柔肝方高剂量组与鳖甲煎丸组疗效最为显著;(2)与空白组比较,模型组miR-23a-3p表达显著降低(P<0.05),芪甲柔肝方高剂量组与鳖甲煎丸组能显著升高miR-23a-3p的表达(P<0.05);(3)miR-23a-3p的靶基因主要参与调控p53、磷脂酰肌醇等通路。结论:芪甲柔肝方治疗CCl4+乙醇的复合造模诱导的大鼠肝纤维化,可显著改善肝功能,降低血清纤维化标志物,减轻肝组织的炎症及纤维化程度,其作用机制可能与上调miR-23a-3p的表达,调控p53、磷脂酰肌醇等通路有关。
Objective: To study the efficacy and anti-fibrosis mechanism of Radix Astragali and Carapax Trionycis Formula(RCF) by method of reinforcing the healthy qi and promoting blood circulation. Methods: The model of liver fibrosis in rats was established by subcutaneous injection of CCl4 peanut oil+10% alcohol in place of water. After judging the success of modeling, the rats were divided into control group, model group, RCF high-dose, medium-dose, low-dose group and Biejiajian Wan group. Gavage treatment was performed separately. The control and the model group were given saline, RCF highdose, medium-dose and low-dose group were treated with RCF decoction, and the Biejiajian Wan group was given Biejiajian Wan suspension. After 6 weeks of treatment, the rats were sacrificed, and liver function, serum liver fibrosis markers and were measured, and the expression of miR-23 a-3 p in liver tissue of each group was detected by RT-qPCR technology. The bioinformatics technique was used to predict the target genes of miR-23 a-3 p and analyze their functions. Results:(1)Compared to the control group, the model group displayed significant higher index of ALT, AST, liver index, serum fibrosis markers(PⅢP, PCIV, HA, LN)(P<0.05) and sever hepatic inflammation and fibrosis. A large number of pseudo lobules were observed in the model group. While all of the RCF high-dose, medium-dose, low-dose group and the Biejiajian Wan group could improve the above index to a certain extent compared with the model group, above which the efficacy of the RCF high-dose group and the Biejiajian Wan group were found the most significant.(2)In comparison with the control group, the expression of miR-23 a-3 p was significantly decreased in the model group(P<0.05), whereas its expression was notably increased in the RCF high-dose group and the Biejiajian Wan group(P<0.05).(3)The results of bioinformatics analysis suggest that miR-23 a-3 p is mainly involved in the regulation of p53, phosphatidylinositol signal pathway. Conclusion: RCF-HD can significantly reduce liver tissue inflammation and fibrosis, and reduce collagen fiber percentage. The anti-fibrosis effect may related to upregulating miR-23 a-3 p and regulate p53, phosphatidyl inositol signals.
作者
苏悦
彭杨芷
刘雨樵
宋虹霏
吴文军
刘西洋
刘毅
冯全生
SU Yue;PENG Yang-zhi;LIU Yu-qiao;SONG Hong-fei;WU Wen-jun;LIU Xi-yang;LIU Yi;FENG Quan-sheng(Chengdu University of TCM,Chengdu 610075,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2021年第7期3962-3966,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
四川省科技计划重点研发项目(No.2020YFS0301)
成都中医药大学“杏林学者”学科人才科研提升计划(No.BSH2020014)。
关键词
扶正通络法
芪甲柔肝方
气虚络阻
肝纤维化
Reinforcing the healthy qi and promoting blood circulation method
Radix Astragali and Carapax Trionycis Formula
Qi deficiency and collateral stagnation
Liver fibrosis