桂枝茯苓胶囊调控mTOR自噬通路改善硼替佐米相关周围神经病变的研究

Mechanism research of Guizhifuling capsule on improving bortezomib-induced peripheral neuropathy by interfering autophagy

目的借助网络药理学及体内外实验探讨桂枝茯苓胶囊(GZFL)干预硼替佐米(Bortezomib,BTZ)导致周围神经病变(Bortezomib-induced peripheral neuropathy,BiPN)的作用成分与机制。方法通过药物-疾病-靶点网络图、交集靶点PPI寻找桂枝茯苓胶囊的有效成分与靶点,并进行GO分析与KEGG通路分析。PC12神经细胞随机分为对照组、模型组及治疗组,免疫荧光检测LC3阳性细胞,Western Blot检测自噬蛋白表达水平。C57BL/6小鼠随机分成对照组、模型组及治疗组,采用BTZ腹腔注射法造模。对各组小鼠进行行为学、组织学检测,应用免疫组化检测小鼠坐骨神经LC3Ⅱ/Ⅰ、Beclin 1、P62、mTOR、NGF表达水平。结果(1)网络药理学分析得到GZFL治疗BiPN的34个有效成分及107个靶点,显著富集于mTOR、MAPK等自噬相关通路;(2)体外实验表明,GZFL能够促进PC12神经细胞自噬水平,减少mTOR表达;(3)体内实验显示,与模型组相比,治疗组小鼠LC3Ⅱ/Ⅰ、Beclin 1、NGF表达水平显著增高,P62、mTOR水平显著下降。结论桂枝茯苓胶囊通过抑制mTOR相关通路促进自噬,改善多发性骨髓瘤硼替佐米治疗导致的周围神经病变,为中医药治疗化疗相关并发症提供新的思路。

Objective To investigate the components and mechanism of Guizhi Fuling Capsule(GZFL)on peripheral neuropathy(BiPN)induced by bortezomib(BTZ)by network pharmacology and in vitro and in vivo experiments.Methods The effective components and targets of Guizhi Fuling Capsule were found by drug-disease-target network diagram and intersection target PPI,and GO analysis and KEGG pathway analysis were carried out.PC12 cells were randomly divided into blank,BTZ,and GZFL groups.LC3 positive cells were detected by immunofluorescence,and the expression level of autophagy protein was detected by Western Blot.C57 BL/6 mice were randomly divided into normal control,model,and GZFL groups.Behavioral and histological tests were performed on each group of mice.Immunohistochemistry was used to detect the expression levels of LC3II/I,Beclin 1,P62,mTOR and NGF in the sciatic nerve of mice.Results 34 effective components and 107 targets of GZFL in the treatment of BiPN were obtained by network pharmacology analysis,which were significantly enriched in autophagy-related pathways such as mTOR and MAPK.In vitro experiments showed that GZFL could promote the level of autophagy and reduce the expression of mTOR in PC12 cells.In vivo experiments showed that compared with the model group,the expression levels of LC3B,Beclin 1 in the GZFL group were significantly increased,and the levels of P62 and mTOR were significantly decreased.Conclusion GuizhiFuling Capsule promotes autophagy by inhibiting mTOR-related pathways,improves peripheral neuropathy caused by bortezomib treatment of multiple myeloma,and provides new ideas for the treatment of chemotherapy-related complications with traditional Chinese medicine.