Study On Anti - Inflammatory And Analgesic Activity

Objective Radix Silenes viscidulae is the dry root of Silene viscidula Franch, which belong to caryophyllaceae Silene. It is used for shortness of breath and cough, cough, phlegm, damp and hot, stomach pain, dyspepsia, abdominal pain, sand urination, rheumatism bone pain, sore furuncle and swollen pain, trauma pain, etc. At Yunnan folk and Yi Medicine, it is a kind of folk medicine with the development and utilization prospects. But on the market, Silene viscidula was confused by its adulterant, not conducive to safety and rational drug use. This article using the method of DNA barcoding to identify Silene viscidula from its varieties or commen adulterants. At the same time, we studied the vivo anti-inflammatory and analgesic activity of n-butanol extract, and the vitro anti-inflammatory activity of n-butyl alcohol as well as monomer compounds, so as to provide the reference of identification, clinical application, and further research of Silene viscidula. Methods In this paper, we using DNA barcode methods to amplify theITS/ITS2 sequence of samples, got BLAST results on the genus, through compare with the inner or interspecific genetic distance of Silene viscidula and Psammosilene tunicoides etc adulterants, build N-J tree for further verify the exactness of the BLAST results; Silene viscidula materials by Et-OH extraction, extraction from n-butyl alcohol, collection of fractions from MCI, D101 macroporous resin, measure and calculate the content of total saponins. Further separate the monomer compounds, make the structure identification according to the relevant data; Give arthritis model rats different doses of n-butanol part drugs, measuring toes swelling and pain threshold value as well as IL1-β、TNF-α content in serum of rats in different time, for the preliminary anti-inflammatory activity mechanism reseach; Further give lipopolysaccharide stimulated, inflammation model cells Raw264.7 different concentrations of n-butyl alcohol parts and monomer compounds, by measuring the cell supernatant of IL1-β、TNF-α content for research the in vitro anti-inflammatory activity of the sample.Results The innerspecific genetic distance of Silene viscidula is less than the interspecific genetic distance, the system tree build successfully consistent with the BLAST results; Total saponins content of Silene viscidula ethanol extract, n-butanol part were 11.32%,18.90%,30%～95% Et-OH fraction of MCI, D101 macroporous resin were 22.05～31.52% and 20.59～32.05% respectively, and isolated a pentacyclic triterpene monomer compounds from the Silene viscidula; Animal experiments show that n-butanol part of high, medium and low dose group of rats foot swelling degrees (375.00±89.491、 359.63±124.24、372.00±138.752μL) compared with model group (504.75±91.802 μL) has significant difference (P<0.05), but the pain threshold of rats has no statistical difference in addition to the medium dose group. High, medium and low dose group of IL1-β content in serum of rats (79.65±15.65、 79.00±14.65、75.96±13.54 pg/mL) compared with model group (79.65±15.65、 79.00±14.65、75.96±13.54 pg/mL), TNF-α content of high, medium and low dose group (23.68±9.02,17.99±10.01、19.96±5.80 pg/mL) compared with model group (40.00±5.32 pg/mL) has very significant difference (P<0.01); Cell inflammation model IL1-β content, n-butanol part of high, medium and low dose group (82.25±7.59、47.19±4.77、56.48±15.10 pg/mL), the low dose group of monomer compound (83.90±12.13 pg/mL) show significant difference (P<0.05) with model group (117.29±35.5 pg/mL), but no statistical difference was found between the TNF-α content.Conclution DNA barcode method can accurately identify Silene viscidula from its adulterants; Silene viscidula n-butanol part has significant anti-inflammatory activity, and anti-inflammatory effect may be related to modulate inflammatory factor IL1-P, TNF-α secretion; Von Frey method show that n-butanol part has no obvious effect on pain threshold of fred’s complete adjuvant rats; n-butyl alcohol and monomer compounds may have play a role on regulating inflammatory cytokines of IL1-β in vitro, and have no influence with the release of TNF-α.