Research on the fingerprint spectrum of salted cypress and prediction analysis of quality markers
Phelllodendron chinense Schneid, a plant in the Rutaceae family, is commonly known as the dried bark of Phelllodendron chinense Schneid. After peeling the bark, remove the rough skin and dry it in the sun. Huangbai has a bitter taste and a cold nature. It has the functions of clearing heat, drying dampness, purging fire, removing steam, detoxifying and treating sores; Salt yellow cypress is a processed product made by roasting yellow cypress with salt water, which can nourish yin and reduce fire. It is used for Yin deficiency and excessive fire, as well as sweating and bone steaming. The main chemical components of Huangbai include alkaloids, limonoids, phenolic acids, and flavonoids. Research has found that there are changes and differences in the chemical composition, content, and pharmacological effects of different processed products of Phellodendron amurense. Salt processed Phellodendron amurense has a better inhibitory effect on Staphylococcus aureus and Corynebacterium diphtheriae than Phellodendron amurense silk. In addition, the metabolic inhibition function of salt processed Phellodendron amurense is superior to that of Phellodendron amurense, which is considered to be the material basis of its nourishing yin effect. Therefore, the nourishing yin effect of Phellodendron amurense is enhanced after salt processing. The 2020 edition of the Pharmacopoeia of the People’s Republic of China (referred to as the “Pharmacopoeia of the People’s Republic of China”) uses HPLC to determine the content of berberine and berberine in salted cypress, and stipulates that their content should not be less than 3.0% and 0.34% respectively. This study takes salted cypress as the research object and establishes 22 batches of salted cypress HPLC fingerprint spectra. Based on principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA) is used to preliminarily screen the differential components of different batches of salted cypress. Combined with fingerprint spectrum data analysis and network pharmacology analysis methods, chemical, biological information, and mathematical statistics are combined. Screening the active ingredients of Platycodon grandiflorus from different angles, and confirming the quality markers of Platycodon grandiflorus, providing a reference for quality control of Platycodon grandiflorus.

 

The chemical components of Huangbai mainly include alkaloids, flavonoids, sterols, and volatile oils, among which alkaloids are the main effective chemical components of Huangbai. After research, it was found that the content of berberine and berberine, the main active ingredients of Huangbai, increased after salt processing. Salt processing is beneficial for the dissolution of its alkaloid components. This study identified four chemical components, namely berberine, berberine, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid, by establishing fingerprint spectra. Through network pharmacology analysis, it was found that berberine and berberine compounds may be effective active ingredients of salt cypress, and potential 8 core targets (PRKCA (degree=10), MAPK14 (degree=6), ADRB (degree=5), AR (degree=4), ROCK1 (degree=4), ROCK2 (degree=4), RAC1 (degree=4), JUN (degree=4)) were screened out. These targets may act on the pathways in cancer, cAMP signaling pathway, cGMP PKG signaling pathway), and cGMP PKG signaling pathway. The TRP channel plays an important role in the regulation of inflammatory mediators of TRP channels, which may be related to the medicinal effects of salt yellow cypress in nourishing yin and reducing fire. Therefore, it is preliminarily predicted that berberine and berberine are potential quality marker components of salt yellow cypress. Modern research results have shown that berberine has pharmacological activities such as antibacterial and anti-inflammatory effects. It can produce anti-inflammatory effects by reducing the expression levels of phosphorized ERK/JNK proteins. It can also inhibit the deubiquitination activity of COP9 signal transducer 5 (CSN5) in non-small cell lung cancer, reduce the expression of programmed cell death ligand 1 (PD-L1), and promote anti-tumor immunity. In addition, berberine also has hypoglycemic, antihypertensive, intestinal tension and amplitude enhancement effects, as well as penetration effects on the prostate. Huangbai alkaloid can inhibit the expression of proteins related to the inflammatory pathway caused by ROS, suppress the IKK/NF – κ B pathway, and significantly reduce the expression of COX-2, a downstream protein of NF – κ B. It has good antioxidant and anti-inflammatory effects both in vivo and in vitro. Quinic acid derivatives exhibit significant activity in anti infection, anti cardiovascular disease, and hypoglycemic effects. Abioylquinic acid compounds are effective phenolic antioxidants with stronger antioxidant capacity than caffeic acid, para hydroxybenzoic acid, ferulic acid, syringic acid, etc. They also have the ability to eliminate free radicals, inhibit mutations, and have anti-tumor effects in the body.
The chemical components of traditional Chinese medicine are complex and diverse, and the quality of traditional Chinese medicine is the basis for its clinical safety and effectiveness. By detecting all the chemical components contained in traditional Chinese medicine through high-performance liquid chromatography, and then using network pharmacology methods, the active compounds contained in it can be quickly obtained. The active ingredients can be efficiently studied, and irrelevant or toxic ingredients can be excluded. The activity of each component on multiple targets can be studied, thereby effectively saving time and cost, and improving the efficiency of screening active ingredients in traditional Chinese medicine. In order to clarify the quality markers of salted cypress, this study used HPLC to study 22 batches of salted cypress from different origins, established fingerprint spectra of salted cypress, confirmed 9 common peaks, and evaluated the similarity of fingerprint spectra of different batches of salted cypress. The similarity was found to be between 0.958 and 0.999, indicating that the chemical composition similarity of the 22 batches of salted cypress slices was high and the quality was relatively stable and uniform. According to the results of principal component analysis, the comprehensive ranking of principal components in Sichuan Province’s decoction pieces is higher than that of other provinces, indicating the scientific nature of Sichuan Province as a traditional authentic production area for Huangbai; OPLS-DA analysis found that berberine and berberine are important differential components of different salt treated Phellodendron amurense slices. Combined with network pharmacology, berberine and berberine are the main active ingredients that exert the efficacy of salt treated Phellodendron amurense. From this, it can be confirmed that berberine and berberine are potential quality markers for salt yellow cypress.

This study used HPLC combined with chemical stoichiometry and network pharmacology to predict berberine and berberine as the active ingredients and quality markers of Platycodon grandiflorus from the perspective of effectiveness, providing reference for the quality control of Platycodon grandiflorus.