Tong Heng-Li, Chen Hao, Gong Fei-Peng, Zhong Ling-Yun, Zhu Jing, Yang Song-Hong
Laboratory of Traditional Chinese Medicine Preparation, School of Pharmacy, Jiangxi University of Traditional Chinese Medicine, Nanchang, China.
Department of Orthopedic, Jiangxi Provincial People's Hospital, Nanchang, China.
Front Pharmacol. 2021 Jun 28;12:680640. doi: 10.3389/fphar.2021.680640. eCollection 2021.
According to the treatment records of Yang deficiency syndrome (YDS) with characteristic decoction pieces of lateral root of -Yinfupian (YF) in traditional Chinese medicine prepare school, known as "Jianchangbang". The aim of this study was to investigate differences in the composition and therapeutic mechanism of the unprocessed lateral root of (ULRA) and its processed product (YF). Ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry and orthogonal partial least squares discriminant analysis method were used to determine and screen the main components of ULRA and YF. Changes in the histological structure and morphology of gonads in rats were observed using hematoxylin-eosin. Enzyme-linked immunosorbent assay was used to determine the contents of serum cyclic adenosine monophosphate and cyclic guanosine monophosphate in YDS rats treated with ULRA and YF. Tandem mass tag proteomics analysis was used to identify the differentially expressed proteins in YDS rats treated with ULRA and YF. Both ULRA and YF exerted certain therapeutic effects on rats with YDS. They improved the gonadal morphology and increased the contents of serum cyclic adenosine monophosphate and cyclic guanosine monophosphate. After processing of ULRA into YF, the content of C19-diester-diterpenoid alkaloids decreased (converted into C19-monoester-diterpenoid alkaloids and C19-alkylol amine-diterpenoid alkaloids), whereas that of C20-diterpene alkaloids increased. Proteomics analysis showed that cytochrome P450 and aldehyde oxidase 3 (AOX3) were downregulated, whereas cathepsin G (CTSG) was upregulated in rats with YDS. Treatment with ULRA mainly downregulated the expression of α-actinin, fast skeletal troponin, creatine kinase, and myosin. Treatment with YF mainly upregulated the expression of mitochondrial ribosomal protein and mitochondrial inner membrane protein. ULRA and YF exerted good therapeutic effects on YDS; the main difference in components between these preparations was in C19-diterpenoid alkaloids. ULRA mainly acts on the muscle contraction-related proteins and is closely related to inflammation and myocardial injury. YF mainly acts on the mitochondrial proteins and is closely related to adenosine triphosphate energy metabolism.
根据中医炮制流派“建昌帮”中以阴附片（YF）的特色饮片治疗阳虚证（YDS）的治疗记录。本研究的目的是探讨附子生品（ULRA）及其炮制品（YF）的成分差异和治疗机制。采用超高效液相色谱-四极杆飞行时间质谱联用和正交偏最小二乘法判别分析方法，对ULRA和YF的主要成分进行测定和筛选。采用苏木精-伊红染色观察大鼠性腺组织结构和形态的变化。采用酶联免疫吸附测定法测定ULRA和YF治疗YDS大鼠血清中环磷酸腺苷和环磷酸鸟苷的含量。采用串联质量标签蛋白质组学分析方法，鉴定ULRA和YF治疗YDS大鼠中差异表达的蛋白质。ULRA和YF对YDS大鼠均有一定的治疗作用。它们改善了性腺形态，增加了血清环磷酸腺苷和环磷酸鸟苷的含量。ULRA炮制为YF后，C19-二酯二萜生物碱含量降低（转化为C19-单酯二萜生物碱和C19-烷醇胺二萜生物碱），而C20-二萜生物碱含量增加。蛋白质组学分析表明，YDS大鼠中细胞色素P450和醛氧化酶3（AOX3）表达下调，而组织蛋白酶G（CTSG）表达上调。ULRA治疗主要下调α-辅肌动蛋白、快肌肌钙蛋白、肌酸激酶和肌球蛋白的表达。YF治疗主要上调线粒体核糖体蛋白和线粒体内膜蛋白的表达。ULRA和YF对YDS均有良好的治疗作用；这两种制剂的主要成分差异在于C19-二萜生物碱。ULRA主要作用于肌肉收缩相关蛋白，与炎症和心肌损伤密切相关。YF主要作用于线粒体蛋白，与三磷酸腺苷能量代谢密切相关。
