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The active ingredients of Ficus hirta and Hypericum perforatum were collected from Traditional Chinese Medicine Database and Analysis Platform(TCMSP) and related papers. The potential targets of these two medicinal herbs were searched from HERB database, and those associated with microvascular angina were screened out from GeneCards, Online Mendelian Inheritance in Man(OMIM), Therapeutic Target Database(TTD), and HERB. Cytoscape was used to construct a protein-protein interaction(PPI) network of the common targets shared by the two herbs and microvascular angina based on the data of String platform. Metascape was employed to identify the involved biological processes and pathways enriched with the common targets. Cytoscape was used to draw the "active ingredient-target-pathway" network. AutoDock Vina was used to dock the core ingredients with the key targets. A total of 19 potential active ingredients and 71 potential targets were identified to be associated with microvascular angina. Bioinformatics analysis showed that phosphatidylinositol-3-kinase/protein kinase B(PI3 K-AKT), interleukin-17(IL17), hypoxia-inducible factor 1(HIF-1) and other signaling pathways were related to the treatment of microvascular angina by F. hirta and H. perforatum. Molecular docking results showed that β-sitosterol, luteolin and other ingredients had strong affinity with multiple targets including mitogen-associated protein kinase 1(MAPK1), epidermal growth factor receptor(EGFR) and so on. These findings indicated that F. hirta and H. perforatum may regulate PI3 K-AKT, IL17, HIF-1 and other signaling pathways by acting on multiple targets to alleviate oxidative stress, inhibit inflammatory response, regulate angiogenesis, and improve vascular endothelium and other functions. This study provides reference for in vitro and in vivo studies of the treatment of microvascular angina.
Subject(s)
Humans , Drugs, Chinese Herbal/pharmacology , Ficus , Hypericum , Medicine, Chinese Traditional , Microvascular Angina , Molecular Docking Simulation , Network PharmacologyABSTRACT
Objective: To control the quality of Fici Hirtae Radix by establishing quality standards. Methods: The origin mistakes of Fici Hirtae Radix in Chinese Pharmacopoeia (1977 edition) were modified by searching and mining plant morphology characters, distribution of origin, historical evolution, clinical efficacy and modern research, medicinal standards in different regions on Ficus hirta Vahl and Ficus simplicissima. Morphological identification, microscopical characteristics based on transverse section and powder, and thin layer chromatography were adopted to identify commercial medicinal materials of Fici Hirtae Radix from different sources. The content of psoralen and bergapten was determined by HPLC. Moisture content, total ash, acid-insoluble ash, and ethanol-soluble extractives were determined by methods described in appendices of Chinese Pharmacopoeia (2010 edition). Results: The origin of Fici Hirtae Radix was not F. simplicissima Lour., but F. hirta Vahl. This study collected 20 Fici Hirtae Radix commercial medicines mixed 40% non-medicinal parts samples. TLC data, morphological characteristics and microscopic characteristics, both showed obvious distinction. The concentration of psoralen and bergapten showed the content of psoralen range from 0.00848% to 0.150%, and the content of bergapten range from 0.00587% to 0.026 3%, and the contents of two index components of Fici Hirtae Radix were significantly higher than the non-medicinal parts samples. The moisture content ranged from 4.73% to 8.27%, total ash ranged for 2.58% to 4.37%, acid insoluble ash ranged for 0.054% to 0.710%, ethanol-soluble extracts ranged for 6.23% to 8.34%. Conclusion: This paper revised the origin error of Fici Hirtae Radix of Chinese Pharmacopoeia (1977 edition) that its specification medicinal part should be the roots of F. hirta, and established quality standards of Fici Hirtae Radix, and these methods could comprehensively and effectively control the quality of commercial medicinal materials of Fici Hirtae Radix. This study could provide evidence for rational, safe, and effective clinical application of Fici Hirtae Radix.
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Objective To study the effects of drug-containing serum of Ficus Hirta on oxidative damage of spleen lymphocyte due to aging in aged mice; To discuss its mechanism of action.Methods Forty aged mice were randomly divided into control group and high-, medium- and low-dose Ficus Hirta groups. Control group was given 0.9% sodium chloride solution for gavage, while high-, medium- and low-dose Ficus Hirta groups were given 6.6, 4.4, and 2.2 g/kg aqueous extract of Ficus Hirta for gavage. The spleen index was observed for optimum dose in aged mice. The optimum time and dilution of drug-containing serum of Ficus Hirta were confirmed by MTT method in lymphocyte proliferation test. The positive rate of senescent cells, the activity of T-SOD and the contents of MDA and ROS were determined in cellular antioxidant experiment after treated by optimal drug-containing serum for 48 h. Results Compared with the control group, the spleen index was significantly improved in high-, medium- and low-dose Ficus Hirta groups (P<0.05,P<0.01). 20% drug-containing serum of Ficus Hirta cultivated for 48 h had the best effects on lymphocyte proliferation in aged mice. 20% drug-containing serum of Ficus Hirta could significantly decrease the positive rate of senescent cells (P<0.01), improve T-SOD activity and decrease the contents of MDA and ROS (P<0.05,P<0.01).Conclusion The drug-containing serum of Ficus Hirta can improve the proliferative activity of spleen lymphocyte in aged mice and the mechanism of action may be involved in decreasing the positive rate of senescent cells and increasing antioxidant ability of lymphocyte.
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Objective: To solve the shortage problem of large-scale planting the seedlings in the roots of Ficus hirta by rapid propagation in vitro tissue culture. Methods: The MS and 1/2 MS media were used as basic media, The multi factor combination of plant growth regulator (6-BA, NAA, 2, 4-D, IAA, KT, and IBA) on the seedling subculture and root culture was studied by orthogonal design. Results: The best medium for the adventitious bud induction was MS + 6-BA 1.0 mg/L + NAA 0.3 mg/L, 72 adventitious buds were obtained by differentiation with 20 d induction of explants; The best medium for cluster inducing and subculture was MS + 1.0 mg/L 6-BA + 0.3 mg/L IAA + 0.3 mg/L KT; The best rooting medium was 1/2 MS + 1.0 mg/L IBA + 0.3 mg/L NAA, and the rooting rate was 100%. Being suitably Transplanted to peat-perlite (1:1) matrix, the 30 d survival rate was 93%. Conclusion: The tissue culture for the roots of F. hirta could be used to produce test-tube seedlings for large scale planting.
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Objective: To clarify the origin of Ficus hirta and to control the quality of the crude drug. Methods: The reference on F. hirta, the microscopic features of powder and tissue of the crude drug, plant morphology, and phytochemical TLC were observed and studied. Results: The result of study provides the using history and experiment research of F. hirta and characteristics of the crude drug, which could be used for the identification of the crude drug and Chinese patent medicine. Conclusion: The convenient and effective method for identification of crude drug and patent drug has been established, which provides the basis for quality control.
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Objective To establish a HPLC method for the determination of psoralen and bergapten in Ficus hirta Vahl.Methods A Merck-lichrospher C18(4 mm?250 mm,5 ?m)column was adopted.The mobile phase was acetonitrile-water(35∶65) with the flow rate of 1.0 mL/min.The column temperature was 35 ℃,and the detection wavelength was set at 222 nm.Results Psoralen's linearity was obtained in the range of 0.12 ?g～1.20 ?g(r=0.999 7),and bergapten's linearity in the range of 0.03 ?g～0.30 ?g(r=0.999 5).Psoralen's average recovery was 100.9 %with RSD 2.9 %,and that of bergapten was 99.6 %with RSD 1.9 %.Conclusion This is the first report of simultaneous determination of psoralen and bergapten in Ficus hirta Vahl.by HPLC,and the results showed the method is accurate,reproducible,and can serve as quality evaluation method for Ficus hirta Vahl.