1.Effects of Different Honey Sunburn Methods on Calycosin and Formononetin in Hedysari Radix
Jiangtao NIU ; Rui CAO ; Xinlei SI ; Tiantian BIAN ; Yuefeng LI
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(9):59-62
Objective To compare the effects of different methods of honey sunburn on the contents of calycosin and formononetin in Hedysari Radix; To provide the basis for the establishment of the optimum processing technology. Methods By frying (traditional method), baking, and microwave methods put Hedysari Radix under honey sunburn. Agilent HC-C18 column (4.6 mm × 250 mm, 5 μm) was used; the mobile phase was acetonitrile-0.01% phosphoric acid solution, gradient elution (0–12 min, 30%–33% acetonitrile; 12–13 min, 31%–40% acetonitrile; 13–25 min, 40% acetonitrile) with velocity of 1.0 mL/min; detection wavelength was 248 nm;the column temperature was 30 ℃; sample volume was 10 μL. Results There was statistical significance in the contents of calycosin and formononetin of different methods of honey sunburn for Hedysari Radix. Among them, the contents of calycosin and formononetin in Hedysari Radix processed by honey roast were the highest, 7.9116 and 49.6996 μg/g, respectively; the contents of calycosin and formononetin in Hedysari Radix processed by traditional method were the lowest, 4.7767 and 37.2910 μg/g, respectively; the contents of calycosin and formononetin in raw Hedysari Radix and Hedysari Radix by honey microwave method were the same, 5.0802, 42.7989 μg/g, and 3.9839, 42.3145 μg/g, respectively. Conclusion Different honey sunburn methods for the contents of calycosin and formononetin in Hedysari Radix have certain effects, and honey roast method is the optimum method.
2.Gene expression profile in human cervical epithelial cells infected with Chlamydia trachomatis
Ru JIA ; Chenli SI ; Mingyang LI ; Jia YANG ; Xinlei WU ; Shanli ZHU
Chinese Journal of Microbiology and Immunology 2023;43(2):93-101
Objective:To compare gene expression profiles in normal human cervical epithelial cells (HcerEpic) before and after Chlamydia trachomatis ( Ct) infection. Methods:HcerEpic cells that were pretreated with DEAE-D were infected with Ct serotype E standard strain and then cultured for 44 h. Uninfected HcerEpic cells were used as the control group. Total RNA was extracted from the cells in each group and reverse transcribed to construct a cDNA library. Differences in gene expression profiles between the two groups were analyzed by high-throughput sequencing and the representative genes were selected for verification by qPCR. Results:A total of 23 997 genes were detected, including 125 differentially expressed genes. Among the 125 genes, 119 were up-regulated and six were down-regulated. GO analysis showed that the differentially expressed genes were enriched in several biological processes including defense response to virus, typeⅠinterferon signaling pathway and cellular responses to typeⅠinterferons. KEGG enrichment analysis showed the differentially expressed genes were mainly enriched in the pathways related to virus infections, such as influenza A virus, herpes simplex virus, EB virus and HPV, and NOD-like receptor pathway.Conclusions:There were significant differences in transcriptome profiles of HcerEpic cells before and after Ct infection. The differentially expressed genes were mainly enriched in the interferon pathway, which was closely related to the antiviral processes in cells. qPCR verified the differentially expressed genes and the genes closely related to the interferon pathway, such as ISG15, IFIT2, OASL and UBE2L6.
3.Optimization of processing technology of honey barked Hedysari Radix using orthogonal design method
Yuefeng LI ; Jiangtao NIU ; Rui CAO ; Xinlei SI ; Tiantian BIAN ; Ke Xing YAN
Journal of Beijing University of Traditional Chinese Medicine 2017;40(2):166-171
Objective To optimize the processing technology of honey baked Hedysari Radix, so as to provide the experimental basis for the quality control of Hedysari Radix processed products and standardization of the processing.Methods L9 (34) orthogonal design method was used, and the mean content of calycosin and formononetin which were detected by HPLC with gradient elution were taken as the comprehensive evaluation indexes, the processing technology was optimized.Results The optimum processing technology of honey baked Hedysari Radix was thickness of 3 cm at the temperature of 70 ℃ for 2.5 h.Conclusion Hedysari Radix baking processed products produced by the optimum technology met with the requirement of Chinese Pharmacopoeia (2015 Edition), which was simple, feasible, and repeatable.The method would provide evidence for further research on optimum processing technology and for standardiztion of industrial production.
4.Datura metel L.inhibits the secretion of inflammatory factors and angiogenesis in keratinocytes in vitro
Boping ZHANG ; Xinlei SI ; Fenfang WU
Basic & Clinical Medicine 2024;44(6):816-820
Objective To investigate the potential role of the traditional herbal medicine Datura metel L.in the treatment of psoriasis using TNF-α-induced inflammation in keratinocytes as a model.Methods Keratinocyte cell line HaCaT was used to establish a psoriasis cell model by tumor necrosis factor-alpha(TNF-α)treatment.The ex-periment comprised three groups:a blank control group,TNF-α-induced psoriasis model group,and TNF-α+Datura metel L.intervention group.Level of IL-17 and CCL20 was measured ELISA,expression of nuclear factor kappa B(NF-κB)subunit p65 protein was measured by Western blot.Endothelial cell tube formation experiment was conducted using an in vitro angiogenesis analysis kit.Results Compared to the TNF-α-induced psoriasis model group,the Datura metel L.extract significantly reduced the levels of IL-17 and CCL20 in the cell culture su-pernatant of TNF-α-induced psoriasis model(P<0.001);Datura metel L.extract markedly decreased the NF-κB subunit p65 protein level in TNF-α-induced psoriasis model cells(P<0.01);Datura metel L.extract effectively inhibited the induction of endothelial cell tube formation by the cell culture supernatant of the psoria-sis model group.Conclusions The Datura metel L.extract down-regulates NF-κB signaling pathway mole-cules,reducing the production of IL-17 and CCL20 inflammatory factors in inflammatory keratinocytes and in-hibiting angiogenesis.