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ObjectiveTo investigate the therapeutic effects and difference in the effects of Arisaematis Rhizoma （AR） before and after processing （i.e.， Arisaematis Rhizoma Preparatum， ARP） with Zingiberis Rhizoma Recens-Alumen on allergic asthma in rats and to provide a basis for the theory of processing improving the efficacy. MethodA rat model of allergic asthma was established in 70 SD rats by intraperitoneal injection of ovalbumin （OVA）-aluminum hydroxide. The rats were administrated with the aqueous extracts of AR （1.2， 0.3 g∙kg-1） and ARP （1.2， 0.3 g∙kg-1） aqueous extracts by gavage， and montelukast sodium （0.001 g∙kg-1） was used as the positive drug. The T helper cell type 1/type 2 （Th1/Th2） ratio in the serum and bronchoalveolar lavage fluid （BALF） and percentages of inflammatory cells in BALF were determined. Polymerase chain reaction （PCR） was employed to determine the mRNA level of mucin 5AC （MUC5AC） in the lung tissue. The pathological changes in the lung tissue were observed by hematoxylin-eosin （HE） staining and PAS staining. Immunohistochemical assay was employed to measure the expression of c-Jun amino-terminal kinase （JNK）， extracellular signal regulated protein kinase （ERK）， and p38 mitogen-activated protein kinase （p38 MAPK） in rat lung tissue. Western blot was employed to determine the protein levels of ERK， p-ERK， JNK， p-JNK， p38， p-p38 in the lung tissue. The effects of AR and ARP were compared based on overall desirability. ResultCompared with the blank group， the levels of interleukin-12 （IL-12） and γ interferon （IFN-γ） in serum and BALF of rats in the model group were significantly lower （P<0.05， P<0.01）， and the levels of interleukin-4 （IL-4）， interleukin-5 （IL-5） and interleukin-13 （IL-13） were significantly higher （P<0.05， P<0.01）. Compared with the model group， the serum and BALF contents of IL-12 and IFN-γ in rats in the montelukast sodium group， high-dose AR group and high-dose ARP group were significantly higher （P<0.05， P<0.01）， and the contents of IL-4， IL-5 and IL-13 were significantly lower （P<0.05， P<0.01）， and the serum contents of IFN-γ in rats in the low-dose AR group and low-dose ARP group were in BALF was significantly higher （P<0.05） and IL-4 and IL-13 were significantly lower （P<0.05， P<0.01）， the percentages of macrophages， lymphocytes， neutrophils， and eosinophils were reduced in BALF， and the expression of JNK/ERK/p38 MAPK signaling pathway and MUC5AC protein was inhibited in lung tissues. Overall assessment of the normalized analysis revealed that the ARP group was slightly more potent than the AR group after administration of the same dose. ConclusionAR and ARP can effectively treat allergic asthma by inhibiting JNK/ERK/p38 MAPK signaling pathway， and the effect is better after concoction， which can provide data support for its "concoction efficiency".
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ObjectiveIn recent years， with the sharp decline of wild resources in Arisaematis Rhizoma and Pinelliae Rhizoma and the immaturity of medicinal cultivation technology， their adulterants have appeared frequently in the market， and the main identifying characteristics have mostly disappeared in the circulation of medicinal materials. Therefore， there is an urgent need to establish a molecular identification method that can quickly and effectively identify the specificity of Arisaematis Rhizoma and Pinelliae Rhizoma. MethodAfter comparison of the rbcL sequences of Arisaematis Rhizoma，Pinelliae Rhizoma， and their adulterants， the specific enzyme cleavage sites Hae Ⅲ and Dra Ⅰ of Arisaematis Rhizoma and Pinelliae Rhizoma， respectively， were selected and identified by polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP）. The main system conditions of PCR-RFLP reaction were established and optimized， and their durability and the ability to detect genuine， adulterants， and mixed counterfeits were investigated. ResultThe PCR-RFLP identification method of Arisaematis Rhizoma and Pinelliae Rhizoma was established. After specific primer amplification， Arisaematis Rhizoma and Pinelliae Rhizoma could be digested by Hae Ⅲ and Dra Ⅰ-restricted endonucleases respectively， at annealing temperature of 54 ℃， the number of cycles of 35， and the amount of DNA template of 3-30 ng， producing two fragments or small cut fragments with a single band between 100-250 bp， whereas the mixed counterfeits were not cleaved and both showed a band at 250 bp. The method is highly accurate in identifying adulterants and mixed counterfeits of Arisaematis Rhizoma or Pinelliae Rhizoma. ConclusionThe PCR-RFLP method developed in this study allows for the rapid identification of Arisaematis Rhizoma and Pinelliae Rhizoma.
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Objective: To establish and analyze the HPLC characteristic chromatogram for water extracts of Arisaematis Rhizoma (AR) and its processed products, Arisaematis Rhizoma Preparatum (ARP) and Arisaema Cum Bile (ACB). The research provided reliable method and scientific basis for their quality control. Methods: The separation was performed on an Agilent C18 (200 mm × 4.6 mm, 5 μm) column with gradient elution of 0.2% acetic acid water and 0.2% acetic acid acetonitrile. The similarity was analyzed with software “Similarity Evaluation System for Chromatographic Fingerprint of TCMs (Version 2012)”. The cluster analysis was performed by SPSS 23.0. The principal component analysis was performed by SIMCA 14.1. Results: HPLC characteristic chromatogram for water extracts of AR, ARP, and ACB were established. There were 19, 20 and 13 common peaks in AR, ARP and ACB, respectively. A total of eight characteristic peaks were identified as F1 (xanthine), F3 (uracil), F4 (hypoxanthine), F5 (uridine), F6 (guanosine), F7 (adenosine), F14 (schaftoside), and F16 (isoschaftoside), respectively. The intragroup similarities of AR, ARP, and ACB were all above 0.8 and each was clustered into one type, and the intergroup similarities among AR and its processed products were all below 0.4. The main components of AR were F16 (isoschaftoside), F14 (schaftoside), F17and F15. The main components of ARP were F16 (isoschaftoside) and F1 (xanthine). The main components of ACB were F3 (uracil), F2, F5 (uridine) and F1 (xanthine). Conclusion: The method can effectively identify AR, ARP and ACB, and provide a scientific basis for their quality control.
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Objective To find delayed luminescence parameters that could characterize the cold and hot properties of Arisaematis Rhizoma Preparatum and Arisaema Cum Bile. Methods Delayed luminescence of Arisaematis Rhizoma Preparatum with addition of Scenedesmus sp. within 26 d after decoction was measured in unequal interval, with aim to verify the stability of the natural delayed luminescence average strength and the linear fitting slope value (k) of excitation delayed luminescence. The delayed luminescence of six batches of Arisaematis Rhizoma Preparatum and Arisaema Cum Bile was measured using biological indicator method, and the content of β-sitosterol in Arisaematis Rhizoma Preparatum and β-sitosterol, bilirubin, and cholic acid of Arisaema Cum Bile was determined using high performance liquid chromatograph (HPLC) to analyze the correlation of k value and the above components content of Arisaematis Rhizoma Preparatum and Arisaema Cum Bile. Results K value of excitation delayed luminescence within 14 d after decoction was steadier than natural delayed luminescence average strength, and k values of six batches of Arisaematis Rhizoma Preparatum were all higher than that of Arisaema Cum Bile. A significant negative correlation between β-sitosterol contents and k values of six batches of Arisaematis Rhizoma Preparatum and Arisaema Cum Bile was found, and no significant negative correlation between bilirubin and cholic acid contents and k values of Arisaema Cum Bile was found. Conclusion K value of excitation delayed luminescence could indicate the differences of medicinal properties of Arisaematis Rhizoma Preparatum and Arisaema Cum Bile, which provides a new method for the study of medicinal properties of Chinese materia medica.