Effect of sulfur fumigation on quality and safety of Lilii Bulbus / 中国中药杂志

Lilii Bulbus is a commonly used Chinese herbal medicine with both medicinal and edible values, while the market products usually has the problem of sulfur fumigation. Therefore, the quality and safety of Lilii Bulbus products deserve attention. In this study, ultra-high performance liquid chromatography-time of flight-tandem mass spectrometry(UPLC-Q-TOF-MS/MS) was combined with principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) to analyze the differential components of Lilii Bulbus before and after sulfur fumigation. We identified ten markers generated after sulfur fumigation, summarized their mass fragmentation and transformation patterns, and verified the structures of phenylacrylic acid markers of sulfur fumigation. At the same time, the cytotoxicity of the aqueous extracts of Lilii Bulbus before and after sulfur fumigation was evaluated. The results showed that in the concentration range of 0-800 mg·L~(-1), the aqueous extract of Lilii Bulbus after sulfur fumigation had no significant effect on the viability of human liver LO2 cells, human renal proximal tubular HK-2 cells, and rat adrenal pheochromocytoma PC-12 cells. Moreover, the viability of the cells exposed to the aqueous extract of Lilii Bulbus before and after sulfur fumigation showed no significant difference. This study identified phenylacrylic acid and furostanol saponins as markers of sulfur-fumigated Lilii Bulbus for the first time, and made clear that proper sulfur fumigation of Lilii Bulbus would not produce cytotoxicity, providing a theoretical basis for the rapid identification and quality and safety control of sulfur-fumigated Lilii Bulbus.

Comparison of Effect of Ancient and Modern Processing Methods on Chemical Components of Lilii Bulbus Decoction Based on UHPLC-Q-Orbitrap HRMS / 中国实验方剂学杂志

ObjectiveTo compare the effects of different processing methods in ancient and modern times on the chemical components of Lilii Bulbus decoction， and to provide experimental support for the origin processing， decoction piece processing and clinical application of this herb. MethodUltra high performance liquid chromatography tandem quadrupole electrostatic field orbitrap high resolution mass spectrometry（UHPLC-Q-Orbitrap HRMS） was used for structural identification of the compounds using excimer ions， secondary MS and characteristic fragment ions， and referring to relevant literature and database information. Principal component analysis（PCA） and orthogonal partial least squares discriminant analysis（OPLS-DA） were used to screen the main differential components， the differential components were quantitatively studied by high performance liquid chromatography（HPLC）， in order to compare the types and contents of chemical components in the decoction of different processing products of Lilii Bulbus. ResultA total of 24 chemical components were identified from the decoction of different processed products of Lilii Bulbus， water extract and scalding liquid of fresh Lilii Bulbus， including 17 phenols， 5 saponins and 2 alkaloids. Compared with the fresh Lilii Bulbus decoction， the contents of regaloside A， p-coumaric acid， colchicine and other components in the decoction of dry Lilii Bulbus processed by scalding method decreased， the content of regaloside C in the decoction of dry Lilii Bulbus processed by steaming method decreased， and the contents of regaloside A and regaloside C in the decoction of fresh Lilii Bulbus processed by water immersion also decreased. Compared with the decoction of dry Lilii Bulbus processed by scalding method， the overall content of components in the fresh Lilii Bulbus decoction and the decoction of fresh Lilii Bulbus processed by water immersion was higher， the contents of components in the decoction of dry Lilii Bulbus processed by steaming method was higher， except for the slightly lower content of regaloside C. ConclusionDifferent processing processes have a certain effect on the types and contents of chemical components in Lilii Bulbus decoction. Scalding process is beneficial to the preservation of Lilii Bulbus， but can cause the loss of effective components. Compared with scalding method， steaming method can prevent browning of Lilii Bulbus and reduce the loss of its active ingredients. The processing method of removing foam after overnight immersion proposed by ZHANG Zhongjing may be more conducive to the treatment of Baihe disease， which can provide reference for the clinical rational application and mechanism research of different processed products of Lilii Bulbus.

Study on mechanism of combination of Platycodonis Radix and Lilii Bulbus with homology of medicine and food in treating pneumonia / 中国中药杂志

To investigate the potential molecular mechanism of the combination of Platycodonis Radix and Lilii Bulbus with the homology of medicine and food in the treatment of pneumonia by means of network pharmacology and in vitro verification experiment. Under the condition of bioavailability(OB)≥30% and drug-like(DL)≥0.18, the active components of Platycodonis Radix and Lilii Bulbus were screened in TCMSP database; the prediction targets of active components were searched from TCMSP, DrugBank and other databases, and the potential targets of pneumonia were obtained through GeneCards and OMIM database. The common targets were obtained by the intersection of drug and disease targets. The PPI network of common targets was constructed by STRING 11.0, and the core targets were obtained by topological analysis. Then the core targets received GO and KEGG analysis with use of WebGestalt and Metascape. The "component-target-pathway" network was constructed with the help of Cytoscape 3.7.1 software, and the component-target molecular docking verification was carried out with Discovery Studio 2016 software. Finally, the core targets and pathways were preliminarily verified in vitro. In this study, 12 active components were screened, 225 drug prediction targets and 420 potential diseases targets were obtained based on data mining method, and 14 core targets were obtained by topological analysis, including TNF, MMP9, AKT1, IL4 and IL2. The enrichment results of GO and KEGG showed that "Platycodonis Radix and Lilii Bulbus" drug pair may regulate inflammation, cell growth and metabolism by acting on 20 key signaling pathways such as TNF and IL-17, thereby exerting anti-pneumonia effects. The results of molecular docking showed that 12 active components had good binding ability with 14 core targets. In vitro experiment results showed that the core components of "Platycodonis Radix and Lilii Bulbus" drug pair could inhibit the expression of MMP9 and TNF-α by regulating TNF signal pathway. This study confirmed the scientificity and reliability of the prediction results of network pharmacology, and preliminarily revealed the potential molecular mechanism of the compatibility of Platycodonis Radix and Lilii Bulbus in the treatment of pneumonia. It provides a novel insight on systematically exploring the mechanism of the compatible use of Platycodonis Radix and Lilii Bulbus, and has a certain reference value for the research, development and application of new drugs.

Theory Analysis on Absorption of Exogenous Substances in Traditional Chinese Medicine / 中国实验方剂学杂志

The safety of traditional Chinese medicine is affected by many factors, and the influence of exogenous harmful substances has been concerned and become a hot spot in recent years, especially heavy metals, pesticide residues and some other harmful substances. In order to explore the effects of non-soil and non-pesticide treatment on residues of these two harmful substances, the heavy metals and agricultural residues of Lilii Bulbus were detected, and the correlation of the data was analyzed. In this experiment, heavy metals and pesticide residues of Scrophulariae Radix were detected, and correlation analysis was conducted for their data. The mechanism of transport phase was interpreted with statistical moment similarity tool of total fingerprint by supramolecular chemistry theory. A large number of experimental data in this paper showed that heavy metals and pesticide contents in Lilii Bulbus basaltifolia had a positive correlation, which was closely related to supramolecular phenomena. Moreover, the similarity of fingerprints between Lilii Bulbus and Scrophulariae Radix suggested that Lilii Bulbus and Scrophulariae Radixa had a high selectivity in absorption of agricultural residues, which proved that the absorption of pesticides in Lilii Bulbus and Scrophulariae Radix from different habitats had the function of supramolecular imprinting template. It was considered that medicinal plant was a giant complex supramolecule with various levels of " imprinted template" . Heavy metals and agricultural residues were also involved in plant growth, forming an " imprinted template" for the formation of supramolecules between agricultural residues and heavy metals. After heavy metals and agricultural residues formed supramolecules, their liposolubility and permeability changed in varying degrees, and their transport in medicinal plants was promoted. Finally, the heavy metal supramolecules of pesticides were absorbed, distributed, aggregated and accumulated in plants. The purpose of the study was to reveal the mechanism of heavy metal and pesticide supramolecule transport, provide a new direction for the treatment of heavy metals and pesticide residues, and ensure the safety of traditional Chinese medicine.

Anti-cancer Mechanism of Polygonati Rhizoma-Lilii Bulbus Based on Network Pharmacology / 中国实验方剂学杂志

Objective::To screen out active ingredients of Polygonati Rhizoma-Lilii Bulbus, and predict the targets and signaling pathways, in order to explore the potential mechanism in treatment of cancer by using network pharmacology. Method::All of active ingredients and targets of Polygonati Rhizoma-Lilii Bulbus were screened out through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). Disease targets for cancer were collected through databases of gene-disease associations (DisGeNET) and Online Mendelian Inheritance in Man (OMIM). Then the Omicshare platform was used to match the active ingredients and the targets for treating cancer. And the " drug-active ingredients-disease targets" network was established using Cytoscape 3.7.0 software. The functional protein association networks (String) database was used to construct the protein interaction network of drug pair targets for treating cancer. Finally, the Functional Annotation Bioinformatics Microarray Analysis (DAVID) database was used to analyze the biological functions and metabolic pathways of key targets. Result::A total of 19 active ingredients were screened out, 234 targets were predicted, 6 active ingredients were identified to be related to cancer. The anti-cancer effect was mainly correlated with the regulation of target proteins in treating cancer, such as Akt serine/threonine kinase 1 (Akt1), Jun proto-oncogene, AP-1 transcription factor subunit (JUN), vascular endothelial growth factor A (VEGFA), matrix metallopeptidase-9 (MMP-9), Caspase-3, Fos proto-oncogene, AP-1 transcription factor subunit (FOS), proteoglycans in cancer, estrogen signaling pathway, human immunodeficiency virus 1 infection, hypoxia inducible factor-1 (HIF-1) signaling pathway, tumor necrosis factor(TNF) signaling pathway, microRNAs in cancer and other pathways. Conclusion::The anti-cancer effect of Polygonati Rhizoma-Lilii Bulbus reflects multi-component, multi-target, multi-pathway characteristics of TCM, and provides a scientific basis for explaining the mechanism and material basis of anti-cancer treatment.

Herbal textual research on triditional Chinese medicine "Baihe"( Lilii Bulbus) / 中国中药杂志

To clarify the species and preparation of " Baihe"( Lilii Bulbus),a traditional Chinese medicine,we investigated the relative ancient Chinese literature on this medicine. The study concluded that Lilium brownii var. viridulum is the authentic lily for medical use. In the Ben Cao Yan Yi and some medical books in the Ming and Qing Dynasties,L. lancifolium( Juandan) was also mistakenly used as genuine lily,but most doctors believe that this variety should not be used for medicinal purposes; L. pumilum( Shandan)began to be used as a medicine from Ri Hua Zi Ben Cao,but mainly for surgery,the effect is also different from L. brownii var. viridulum. We suggested Shandan be used as the species for another medicine as " Hong Bai He( red lily) " due to its red flower. All above three species recorded in the Chinese Pharmacopoeia are not corresponded to the condition of ancient doctors' uses. Therefore,as for developing of traditional classical formula,L. brownii var. viridulum should be chosen and used as Baihe. The birth places for Baihe include Gansu,Hubei,Anhui and Shandong province. The drug preparations of Baihe include crude medicine,roasting and steaming,which preparation should be chosen depends on the formula which contains Baihe.

Post Harvest Investigation on Processing,Packaging,and Storage of Lilii Bulbus / 中国实验方剂学杂志

Objective: To investigate the current harvest,processing,packaging and storage of Lilii Bulbus due to the great difference in quality of commercially available products,in order to realize the factors affecting the quality after harvest and provide reference for the standardized production. Method: A series of factors affecting Lilii Bulbus quality were analyzed comprehensively on the basis of both field survey in main producing areas and medicinal material markets and literature retrieval. Result: The current situation and problems in harvest,initial processing,drying technologies,packaging,and storage were summarized. Conclusion: The harvest link of Lilii Bulbus involves growing years,harvesting time,and collecting methods. The initial processing link involves stripping,cleaning,sulfur fumigation,and many cooking and drying methods. The hot-air,vacuum-freeze,far-infrared,and microwave methods are applied in drying Lilii Bulbus. The main metamorphic phenomena are brown staining,moth biting,and decay in the packaging and storage links. Lilii Bulbus are packed mainly in bulk,and a few of them are packed in plastic bags or vacuum bags for fresh bulbus. Because the operation process is not standard,the cooking and drying methods are the key factors affecting the quality of Lilli Bulbus. It is necessary to establish a scientific and unified standard operating procedure (SOP) for controlling the whole process of Lilii Bulbus in multiple links.

Study on effects of total saponins from Lilii Bulbus on proliferation, apoptosis, invasion and metastasis of lung cancer cells and its preliminary mechanism / 中国中药杂志

The potential role of total saponins extracted from Lilium lancifolium bulbs (TSLL) on the proliferation, apoptosis, migration and invasion of human lung cancer A549 cells and its possible mechanism were discussed. Effect of TSLL on proliferation of A549 cells were detected by CCK-8, clone formation assay and EdU staining. Effect of TSLL on apoptosis morphology of A549 cells was observed by fluorescence microscope using Annexin V/PI double staining and Hoechst 33342 staining. Effect of TSLL on cell migration and invasion was detected by Transwell migration test and Transwell invasion test, respectively. Western blot was used to detect TSLL on the expression change of intracellular associated proteins. Results showed that TSLL intervention in A549 cells within 24, 48 or 72 h significantly inhibited cell growth, and its IC₅₀values were about 229, 173 and 71 mg·L⁻¹, respectively. TSLL significantly reduced the clone formation rate of A549 cells and decreased the DNA synthesis rate of A549 cells in a concentration dependent manner. TSLL induced A549 cells apoptosis and reduced the migratory behavior of A549 cells. TSLL decreased invasion of A549 cells to the artificial basement membrane. The expression level of intracellular PCNA and the ratio of Bcl-2/Bax protein were down-regulated and procaspase 3 was activated. In addition, TSLL had no obvious effect on epithelial mesenchymal transition (EMT) related marker proteins E-cadherin and vimentin expression. The above results indicated that TSLL possess inhibitory effects against proliferation, migration and invasion of lung cancer A549 cells and apoptosis-induced effect. The anti-proliferation effect of TSLL is very likely by inhibiting intracellular DNA synthesis through reducing the expression of PCNA in lung cancer cells. And the apoptosis induction of TSLL on lung cancer cells is associated with the regulation of Bcl-2 and Bax proteins expression. Nevertheless, there is no incontestable correlation between anti-invasion and metastasis effects of TSLL and EMT in lung cancer cells.

Textual research on origin and genuine of Lilii Bulbus / 中国中药杂志

Lilii Bulbus, which comes from many medicinal plants，is a frequently-used traditional Chinese medicine，the records in previous herbal literatures of it's origin and quality were inconstant. To trace back it's sources, we conducted a systematical study on it's origin and quality by textual research and investigation in this paper,The result showed that the origins of Lilii Bulbus are mainly source from white-flowers, red-yellow-flowers and red-flowers of Lilium， L. brownii var. viridulum, which were believed authentic or good quality in all previous herbal literatures,and L. pumilum and L. concolor which belong to white-flowers,and L. lancifolium which belong to red-yellow-flowers were believed low-quality and unfit for medicinal uses, or they were listed below and often have different effect with L. brownii var. viridulum. Among them, only L. concolor does not belong to Lilii Bulbus according to Chinese Pharmacopeia (2015 edition), The mainstream varieties of Lilii Bulbus became L. lancifolium now according to our practical investigation, which were very different from previous herbal literatures. Although chemical and pharmacological studies provided a reference for L. lancifolium，we should respect the actual records of the previous herbal literatures，the research of material foundation of efficacy should be more sufficient，and provide science evidence for clinical application of different species of Lilii Bulbus.