A total of 84 species of the plant genus Silene (Caryophyllaceae) are indigenous to Central Asia [1]. We studied previously ecdysteroids from the aerial part of three species of Silene [2][3][4]. Lipids extracted by CHCl 3 during isolation of the ecdysteroids were studied in order to utilize more of the raw material.Ecdysteroids were isolated by soaking the ground air-dried aerial part of the plants in MeOH at room temperature for 1 d. Then, the extract was filtered. The MeOH was distilled in a rotary evaporator. The solid was dissolved in water and treated with CHCl 3 . The ecdysteroids remained in the aqueous layer. The CHCl 3 part of the extract was not used.We isolated the lipids from it using column chromatography over silica gel and solvent system hexane:Et 2 O (1:1). This isolated neutral lipids (NL). Their composition was determined using TLC and hexane:Et 2 O (4:1 and 1:1). The principal components of the NL were triglycerides, free fatty acids, and free sterols. Fatty acids were isolated from NL after their alkaline hydrolysis and were analyzed by GC as the methyl esters on a Chrom 5 instrument with a flame-ionization detector using a metallic column (2.5 m) packed with Chromaton N-AW with Reoplex-400 (15%). Table 1 presents the fatty-acid composition.It can be seen that the qualitative fatty-acid compositions of NL from the aerial parts of the aforementioned species of Silene were identical and consisted of 15 acids. Their quantitative compositions differed. This was evident in the content of the principal acids, the saturated one of which was palmitic acid (16:0); the unsaturated ones, two essential fatty acids Z-6 (18:2) and Z-3 (18:3). Palmitic acid dominated the NL in plants 2 and 3; the sum of linoleic and linolenic acids, in S. viridiflora.Next the antibacterial activity of these CHCl 3 extracts was determined by the disk diffusion test [5]. For this, they were dissolved in DMSO. Microorganisms were grown in agar dishes at 30°C overnight in Mueller-Hinton medium (Oxoid). The suspension (100 PL) contained 10 8 CFU/mL. Sterile filter disks (5-mm diameter) were soaked with extract solution (20 PL, 5 mg/mL) and placed on the surface of the innoculated Petri dishes. The dishes were incubated at 37°C for 24 h. The antibacterial activity was estimated by measuring the inhibition zone formed around the disks. We used five disks in each Petri dish. Each test was performed in triplicate.The test cultures of microorganisms were the following: Klebsiella pneumoniae 40602; Micrococcus luteus obtained from the Microbiology Faculty, Manchester University, GB; Pseudomonas aeruginosa NCTC6749; Enterococcus faecalis NCTC775; Proteus rettgeri NCIMB9570 obtained from the National Collection of Great Britain (NCTC); bacterial strains P. agglomerans T26, B. cereus T80, and Staphylococcus saprophyticus T415 obtained from the Biology Faculty, National University of Uzbekistan.Antimicrobial activity was found by testing against several bacterial cultures. The CHCl 3 extract of S. brachuica inhibited growth of three Gram-...
We have previously reported that 2-deoxyecdysone, 2-deoxy-20-hydroxyecdysone, polipodin B, 20-hydroxyecdysone, 26-hydroxypolipodin B, integristerone A, 2,22-diacetate-and 3,22-diacetate-20,26-dihydroxyecdysone, sileneoside A, and sileneoside D were isolated from the aerial part of Silene viridiflora L. (Caryophyllaceae) introduced into the experimental plot of the ICPS AS RU [1,2]. In continuation of research on the chemical composition of this plant, we isolated fractions containing phytoecdysteroid 1 by chromatography of the BuOH extract over silica gel using CHCl 3 :CH 3 OH (9:1). Fractions were rechromatographed using CHCl 3 :CH 3 OH (15:1) to afford 1. The PMR and mass spectra allowed 1 to be identified as taxisterone, C 27 H 44 O 6 . Mass spectrum (CI, NH 3 , m/z): 465 [M + H] + ,
Investigations of the plants from the genus Silene showed that they are sources of ecdysteroids [1] and that many of them contain significant quantities of biologically active compounds.We studied lipids occurring in the CHCl 3 extract of S. guntensis B. Fedtsch. during extraction from it of ecdysteroids in order to utilize the plant more completely and safely and to establish the effect of the lipid components on the effectiveness of the antibacterial and cytotoxic activity.Neutral lipids were isolated by separating the extract over a column of silica gel into hydrocarbons (3.34%), esters of fatty acids (FAE) and alcohols (7.83), triacylglycerides (TAG) (2.48), free fatty acids (FFA) (3.12), aliphatic and polyprene alcohols (5.0), and compounds eluted by Et 2 O (19.58), CHCl 3 (3.38), and MeOH (55.26).It can be seen that FAE, FFA, and TAG made up 13.43% of the total. Fatty acids were analyzed as the methyl esters using GLC (Table 1) [2]. Table 1 shows that the FAE and FFA were more enriched in saturated acids than TAG and that the TAG have more unsaturated acids. This is typical for these classes of neutral lipids. The principal saturated acid was palmitic (16:0). Moreover, the FAE fraction included a significant quantity of 24:0 and 26:0 acids (21.3% total) whereas the FFA had 10.3% of the 26:0 acid. The dominant unsaturated acids in these lipid classes were oleic (18:1), linoleic (18:2), and linolenic (18:3).Preliminary screening of the CHCl 3 part of the extract from the aerial part of S. guntensis found that it exhibited antibacterial and cytotoxic activities.Cell Cultivation. Antitumor activity of the tested samples was evaluated using HeLa cells that were cultivated at 37°C under an atmosphere with 90% humidity and 5% CO 2 in 96-well plates (2 u 10 4 cells/well) in DMEM culture medium (pH 7.4) containing inactivated fetal calf serum (10%), penicillin/streptomycin (100 U/mL), sodium pyruvate (1 mM), and non-essential amino acids (10 mM). After this, the CHCl 3 extract at a concentration of 0.97-500 Pg/mL was added to the HeLa cells, which were placed in an incubator for 24 h at 37°C. The control was HeLa cells (2 u 10 4 cells/well) without the extract. The results were calculated in percent of the control. All experiments were conducted in triplicate.MTT-test. Cytotoxic activity was evaluated using the MTT test [3]. Medium in each well was replaced 4 h before the end of cultivation by fresh medium (DMEM, 100 PL) containing MTT (0.5 mg/mL). The plates were placed in the incubator for 3 h. The crystals of formazane that were present at the end of the experiment were dissolved in DMSO. Absorption was measured in an apparatus for reading the plates. Cytotoxic activity of the extracts and compounds was determined from the percent reduction in the controls of the color at 595 nm.The preliminary in vitro investigations showed that the CHCl 3 extract of S. guntensis suppressed the growth of HeLa cells. The inhibiting concentration of the CHCl 3 extract according to the MTT tests was a dose of 26.58 Pg/mL. At that ...
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