Hepatoprotective effect of lactic acid bacteria, inhibitors of β-glucuronidase production against intestinal microflora

The hepatoprotective activity of lactic acid bacteria (Lactobacillus brevis HY7401, Lactobacillus acidophilus CSG and Bifidobacterium longum HY8001), which inhibited beta-glucuronidase productivity of intestinal microflora, on t-BHP- or CCl4-induced hepatotoxicity of mice were evaluated. These oral administration of lactic acid bacteria lowered beta-glucuronidase production of intestinal microflora as well as Escherichia coli HGU-3. When lactic acid bacteria at a dose of 0.5 or 2 g (wet weight)/kg was orally administered on CCl4-induced liver injury in mice, these bacteria significantly inhibited the increase of plasma alanine transferase and aspartate transferase activities by 17-57% and 57-66% of the CCl4 control group, respectively. These lactic acid bacteria also showed the potent hepatoprotective effect against t-BHP-induced liver injury in mice. The inhibitory effects of these lactic acid bacteria were more potent than that of dimethyl diphenyl bicarboxylate (DDB), which have been used as a commercial hepatoprotective agent. Among these lactic acid bacteria, L. acidophilus CSG exhibited the most potent hepatoprotective effect. Based on these findings, we insist that an inhibitor of beta-glucuronidase production in intestine, such as lactic acid bacteria, may be hepatoprotective.     

Inhibitory effects ofBifidobacterium Spp. isolated from a healthy korean on harmful enzymes of human intestinal microflora

Five hundreds of bifidobacteria were isolated from a healthy Korean and the inhibitory effects of these isloated bacteria on harmful enzymes of human intestinal microflora were examined by cocultivation of the isolated bifidobacteria withE. coli or total human intestinal microflora. In comparison with the results ofE. coli or intestinal microflora cultivation,Bifidobacterium breve K-110,B. breve K-111 andB. infantis K-525 effectively inhibited harmful enzymes (β-glucuronidase and tryptophanase) of f.coli and lowered the pH of the culture media. Also they inhibited the harmful enzymes (β-glucosidase, β-glucuronidase, tryptophanase and urease) and ammonia production of intestinal microflora, and lowered pH of the culture media by increasing lactic acid bacteria of intestinal microflora. When these isolated bifidobacteria were administered on mice, fecal harmful enzymes were also inhibited. Among tested bifidobacteria,B. breve K-110 had the highest inhibitory effect of fecal harmful enzymes.     

Resistance mechanism ofAcinetobacter spp. strains resistant to DW-116, a new quinolone

DW-116 is a new fluoroquinolone antimicrobial agent with a broad spectrum. In order to elucidate the resistance mechanism to DW-116 inAcinetobacter spp. bacteria, total chromosomal DNA was isolated from 10 strains ofAcinetobacter spp. resistant to DW-116. Quinolone resistance determinant region (QRDR) of DNA gyrase gene was amplified by PCR. The 345 bp nucleotide fragment yielded was inserted into pKF 3 which was used as the vector. Comparisons of the DNA sequences of 8 strains with that of the wild type strain revealed a Ser-83 to Leu mutation in mutants and all ten strains contained one silent mutation(T→G) in QRDR. FromAcinetobacter MB4-8 strain, DNA gyrase was isolated and purified, through novobiocin-sepharose, heparin-sepharose affinity column chromatography. The enzyme was composed of two subunits and the molecular mass of subunits A and B were 75.6 and 51.9 kDa, respectively. The supercoiling activity of the reconstituted DNA gyrase composed of subunit A fromAcinetobacter MB4-8 and subunit B fromE. coli was not inhibited by 128 μg/ml of ciprofloxacin. It might be said that one of the resistance mechanisms to DW-116 inAcinetobacter MB4-8 was subunit A alteration of DNA gyrase.     

Triterpenoic acids of Prunella vulgaris var. lilacina and their cytotoxic activities In Vitro

The column chromatographic separation of the MeOH extract from the aerial parts of Prunella vulgaris var. lilacina Nakai led to the isolation of fifteen triterpenoic acids (2–6, 9–13, 16–20), four flavonoids (14, 21–23), four phenolics (7, 8, 15, 24), and a diterpene (1). Their structures were determined by spectroscopic methods to be trans-phytol (1), oleanic acid (2) ursolic acid (3), 2α,3α,19α-trihydroxyurs-12en-28oic acid (4), 2α,3α-dihydroxyurs-12en-28oic acid (5), maslinic acid (6), caffeic acid (7), phydroxy cinnamic acid (8), 2α,3α,19α,23-tetrahydroxyurs-12en-28oic acid (9), 2α,3α,23-trihydroxyurs-12en-28oic acid (10), 2α,3β-dihydroxyurs-12en-28oic acid (11), 2α,3β,24-trihydroxyolea-12en-28oic acid (12), (12R, 13S)-2α,3α,24,trihydroxy-12,13-cyclo-taraxer-14-en-28oic acid (13), quercertin 3-O-β-D-glucopyranoside (14), rosmarinic acid (15), 2α,3α,24-trihydroxyurs-12,20(30)-dien-28oic acid (16), 2α,3α,24-trihydroxyolea-12en-28oic acid (17), 2α,3β,19α,24-tetrahydroxyurs-12en-28oic acid 28-O-Dglucopyranoside (18), 2α,3α,19α,24-tetrahydroxyurs-12en-28oic acid 28-O-D-glucopyranoside (19), prunvuloside A (20), kaempferol 3-O-α-L-rhamnopyranosyl(1→6)-β-D-glucopranoside (21), kaempferol 3-O-β-D-glucopyranoside (22), quercertin 3-O-α-L-rhamnopyranosyl(1→6)-β-D-glucopyranoside (23), and 2-hydroxy-3-(3’,4’-dihydroxyphenly)propanoic acid (24). Compounds 1, 8–12, 17, 21, 23, and 24 were isolated from this plant source for the first time. The isolated compounds were evaluated for their cytotoxicity against A549, SK-OV-3, SK-MEL-2, and HCT15 cells in vitro using the sulforhodamin B bioassay (SRB) method. Compound 3 exhibited moderate cytotoxic activity against A549, SK-OV-3, SK-MEL-2, and HCT15 cells, with ED₅₀ values of 3.71, 3.65, 13.62, and 5.44 μM, respectively.     

西洋参茎叶总皂苷氧化裂解产物中的新侧链环合型达玛烷型三萜

为了研究西洋参茎叶总皂苷氧化碱解产物的组成，对加拿大产西洋参茎叶总皂苷进行氧化碱解后，其产物通过硅胶柱色谱、Sephadex LH-20柱色谱、重结晶等方法进行分离纯化得到2个化合物。根据化合物的理化性质和光谱数据，鉴定其结构为：(12R,20S,24R)-20,24;12,24-diepoxy-24-deisopropyl-dammarane-3β-ol (1)和(20S,24R)-20,24-epoxydammarane-3β,12β,25-triol (2)。化合物1，2均为首次从西洋参茎叶总皂苷碱解产物中分离得到侧链环合的达玛烷型三萜，其中化合物1为新化合物。     

Aromatic compounds and triterpenoidal saponins fromClematis koreana var.umbrosa

From the methanolic extract of aerial parts ofClematis koreana var.umbrosa, one new triterpenoidal saponin, 3-O-β-D-xylopyranosyl(1–3)-α-L-arabinopyranosyl oleanolic acid 28-O-α-L-rhamnopyranosyl(1–4)-β-D-glucopyranosyl(1–6)-β-D-glucopyranosyl ester, along with five known aromatic compounds and two known triterpenoidal saponins were isolated.     

Constituents of the leaves and roots of Ligularia stenocephala MATSUM. et KOIDZ.

Constituents of the leaves and roots of Ligularia stenocephala MATSUM. et KOIDZ. (Compositae) were investigated. Three compounds, (2E,6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyl-2,6,10,14,18,22-tetracosahexaen-1-ol, (3E,6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyl-3,6,10,14,18,22-tetracosahexaen-2-ol and neophytadiene, were isolated from the leaves of L. stenocephala. Six compounds, 5-acetyl-6-hydroxy-2α-isopropenyl-3β-methoxy-2,3H-benzofuran, 3β-acetoxy-6-acetyl-5-hydroxy-2α-isopropenyl-2,3H-benzofuran, p-hydroxybenzaldehyde, vanillin, 4-hydroxyacetophenone and 4-hydroxy-3-methoxyacetophenone, were isolated from the roots of L. stenocephala. The structures were determined on the basis of spectral data.     

Chemical composition and biological activity of total bufadienolides from the Central-Asian <Emphasis Type="Italic">Bufo viridis</Emphasis> toad venom

The chemical composition of bufadienolides isolated from the venom of Bufo viridis green toad occurring in Central Asia was determined and their biological properties were studied. Six individual bufadienolides were isolated by reverse-phase chromatography on a Lichrosorb RP-8 (10 μ) column in amounts sufficient for qualitative analysis. Two of these were previously identified as arenobufagin and gamabufotalin by NMR spectroscopy and x-ray diffraction methods. The chemical structures of four other bufadienolides are now established by NMR spectroscopy and HPLC. These compounds have been identified as telocinobufagin (3β,5β,14β-trihydroxybufa-20,22-dienolide), marinobufagin (3β, 5β-hydroxy-14,15β-epoxybufa-20,22-dienolide), bufarenogin (3β,12β,14β-hydroxybufa-20,22-dienolide), and bufalin (3β,14β-hydroxybufa-20,22-dienolide). __________ Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. 41, No. 11, pp. 33–37, November, 2007.     

Ginsenoside Rs3, a genuine dammarane-glycoside from Korean red ginseng

A genuine dammarane-glycoside, named as ginsenoside Rs₃, was isolated from the MeOH extracts of Korean red ginseng (Panax ginseng C.A. Meyer) through repeated silica gel column chromatographies and its chemical structure was determined as (20S)-protopanaxadiol 3-O-[6″-O-acetyl-β-D-glucopyranosyl (1→2)-β-D-glucopyranoside on the basis of several spectral and physical evidences including HMBC and FAB-MS.     

Immunosuppressive effects of new cyclolanostane triterpene diglycosides from the aerial part of <Emphasis Type="Italic">Cimicifuga foetida</Emphasis>

Two new cyclolanostane diglycosides, cimifoetiside A (1) and cimifoetiside B (2), were isolated from an 80% ethanolic extract of the aerial part of Cimicifuga foetida L. (Ranuculaceae). Using spectral data and chemical analysis, the structures of 1 and 2 were identified as (23R, 24S) cimicigenol 3-O-β-D-glucopyranosyl-(1″→3′)-β-D-xylopyranoside and (23R, 24S) cimicigenol 3-O-β-D-glucopyranosyl-(1″→2′)-β-D-xylopyranoside, respectively. The in vitro immunosuppressive effects of the two new compounds 1 and 2, as well as four other known cyclolanostane saponins 3–6 on T cells were evaluated. All the agents tested effectively inhibited the proliferation of murine splenocytes induced by Concanavalin A (ConA), with IC₅₀ values ranging from 12.7 nM to 33.3 nM.     

Isolation and identification of chondroitin sulfates from the mud snail

Chondroitin sulfates were isolated from the mud snail. For the quantitative analysis of enzymatic digestion products of isolated chondroitin sulfates, strong anion exchange-high performance liquid chromatography (SAX-HPLC) was performed. By the action of chondroitinase ABC, three unsaturated disaccharides 2-acetamide-2-deoxy-3-O-(β-D-gluco-4-enepyranosyluronic acid)-D-galactose (ΔDi-OS), 2-acetamide-2-deoxy-3-O-(β-D-gluco-4-enepyranosyluronic acid)-6-O-sulfo-D-galactose (ΔDi-6S) and 2-acetamide-2-deoxy-3-O-(β-D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose (ΔDi-4S) were produced from the mud snail chondroitin sulfates. The analysis showed that relative proportion of ΔDi-OS/ΔDi-6S/ΔDi-4S was 58.7/3.1/38.2. The immunomodulating activity of chondroitin sulfate was examined by cell proliferation assay and these results suggest that it might be a immunosuppressant.     

Chemical and biologically active constituents of <Emphasis Type="Italic">Pteris multifida</Emphasis>

A new compound, 4-caffeoyl quinic acid 5-O-methyl ether (2), together with 12 known compounds—identified as (2R,3R)-pterosin L 3-O-β-d-glucopyrannoside (3), β-sitosterol β-d-glucopyranoside (4), apigenin 7-Ο-β-d-glucopyranoside (5), luteolin 7-Ο-β-d-glucopyranoside (6), sucrose (7), caffeic acid (8), pterosin C 3-Ο-β-d-glucopyranoside (9), pteroside C (10), 4,5-dicaffeoyl quinic acid (11), pteroside A (12), wallichoside (13) and (2S)-5,7,3′,5′-tetrahydroxyflavanone (14)—were isolated from Pteris multifida. The structure of the new compound was determined by means of physical, chemical and spectroscopic evidence. Compounds 5 and 6 were the main constituents of the plant, with yields of 0.19% and 0.16%, respectively. The cytotoxic activities of 2, 3, and 9–13 were evaluated against a human cell line (KB cells). Among the isolated compounds, pterosin C 3-Ο-β-d-glucopyrannoside (9) and 4,5-dicaffeoylquinic acid (11) showed a significant selective cytotoxicity (IC₅₀ 2.35 and 5.38, respectively), while moderate activity was observed for compound 2 (IC₅₀ 12.3). The chemosystematics of Pteris species is also discussed.     

Two new glucuronide saponins,Achyranthosides G and H,from <Emphasis Type="Italic">Achyranthes fauriei</Emphasis> root

Two oleanolic acid saponins named achyranthosides G (1) and H (2) were newly isolated from Achyranthes fauriei root as methyl esters in addition to methyl esters of achyranthosides A - F and five oleanolic acid glucuronides (chikusetsusaponins IVa, V, 28-deglucosyl chikusetsusaponin V, pseudoginsenoside RT₁, and oleanolic acid 3-O-β-d-glucuronopyranoside) as well as oleanolic acid 28-O-β-d-glucopyranoside, β-ecdysterone, and polypodine B. Their structures were characterized as follows on the basis of the chemical and spectroscopic evidences.     

Purification and characterization of GTP cyclohydrolase I fromStreptomyces tubercidicus, a producer of tubercidin

GTP cyclohydrolase I catalyzing the first reaction in the biosynthesis of pterin moiety of folic acid in bacteria, was purified fromStreptomyces tubercidicus by at least 203-fold with a yield of 32% to apparent homogeneity, using ammonium sulfate fractionation, DEAE-cellulose, Sepharose CL-6B, and hydroxylapatite column chromatography. The molecular weight of the native enzyme was estimated to be 230,000 daltons by gel permeation chromatography. The purified enzyme gave a single band on sodium dodesyl sulfate-polyacrylamide gel electrophoresis and its molecular weight was apparently 58,000 daltons. These results indicate that the enzyme consists of four subunits with the same molecular weight. TheK _m andV _max values for GTP of the purified enzyme were determined to be 80 μM and 90 nmol/min (mg protein), respectively. The optimum pH and temperature for the enzyme reaction were pH 7. 5∼8.5 and 40∼42°C, respectively. Coenzyme or metal ion was not required for the enzyme activity. The enzyme activity was inhibited by most divalent cations, while it was slightly activated by potassium ion. In case of nucleotides, CTP, GMP, GDP, and UTP inhibited enzyme activity, among which GDP exhibited the strongest inhibitory effect.     

Chlorogenic acid, an antioxidant principle from the aerial parts ofArtemisia iwayomogi that acts on 1,1-diphenyl-2-picrylhydrazyl radical

The antioxidant activity ofArtemisia iwayomogi was determined by measuring the radical scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical The methanol extract ofA. iwayomogi showed strong antioxidant activity, and thus fractionated with several solvents. The antioxidant activity potential of the individual fraction was in the order of ethyl acetate>n-butanol>water>chloroform>n-hexane fraction. The ethyl acetate andn-butanol soluble fractions exhibiting strong antioxidant activity were further purified by repeated silica gel and Sephadex LH-20 column chromatography. Antioxidant chlorogenic acid was isolated as one of the active principles from then-butanol fraction, together with the inactive components, 1-octacosanol, scopoletin, scopolin, apigenin 7,4′-di-O-methylether luteolin 6,3′-di-O-methylether (jaceosidin), apigenin 7-methylether (genkwanin), 2,4-dihydroxy-6-methoxyacetophenone 4-O-β-D-glucopyranoside and quebrachitol. The antioxidant activity of chlorogenic acid was comparable to that of L-ascorbic acid, which is a well known antioxidant.     

Preliminary phytochemical investigation of <Emphasis Type="Italic">Lychnis flos-cuculi</Emphasis> herbs

Five polyphenolic compounds were identified from the diethyl ether and ethyl acetate extracts of the herb of ragged robin, Lychnis flos-cuculi L. (Caryophyllaceae). The identified and isolated compounds were established as a caffeic acid, luteolin, apigenin, apigenin 8-C-β-d-glucopyranoside (vitexin) and luteolin 8-C-β-d-glucopyranoside (orientin) by means of extensive spectroscopic studies, including UV and NMR, and confirmed by their chromatographic behavior—TLC and HPLC analyses. This is the first report of occurrence of these compounds in L. flos-cuculi.     

Flavonol glycosides from the aerial parts of<Emphasis Type="Italic">Aceriphyllum rossii</Emphasis> and their antioxidant activities

The methanol extract obtained from the aerial parts ofAceriphyllum rossii (Saxifragaceae) was fractionated into ethyl acetate (EtOAc),n-BuOH and H₂O layers through solvent fractionation. Repeated silica gel column chromatography of EtOAc andn-BuOH layers afforded six flavonol glycosides. They were identified as kaempferol 3-O-β-D-glucopyranoside (astragalin,1), quercetin 3-O-β-D-glucopyranoside (isoquercitrin,2), kaempferol 3-O-α-L-rhamnopyranosyl (1→6)-β-D-glucopyranoside (3), quercetin 3-O-α-L-rhamnopyranosyl (1→6)-β-D-glucopyrano-side (rutin,4), kaempferol 3-O-[α-L-rhamnopyranosyl (1→4)-α-L-rhamnopyranosyl (1→6)-β-D-glucopyranoside] (5) and quercetin 3-O-[α-L-rhamnopyranosyl (1→4)-α-L-rhamnopyranosyl (1→6)-β-D-glucopyranoside] (6) on the basis of several spectral data. The antioxidant activity of the six compounds was investigated using two free radicals such as the ABTS free radical and superoxide anion radical. Compound1 exhibited the highest antioxidant activity in the ABTS2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging method. 100 mg/L of compound1 was equivalent to 72.1±1.4 mg/L of vitamin C, and those of compounds3 and5 were equivalent to 62.7±0.5 mg/L and 54.3±1.3 mg/L of vitamin C, respectively. And in the superoxide anion radical scavenging method, compound5 exhibited the highest activity with an IC₅₀ value of 17.6 ± 0.3 μM. In addition, some physical and spectral data of the flavonoids were confirmed.     

岗松中的一个新黄酮醇苷类化合物

为了研究岗松的化学成分，采用硅胶、聚酰胺柱层析色谱和重结晶的方法对化合物进行分离纯化，通过理化性质和波谱技术鉴定化合物结构。分离并鉴定了8个化合物：6，8-二甲基山柰酚-3-O-α-L-鼠李糖苷(1)、槲皮素(2)、槲皮素-3-O-α-L-鼠李糖苷(3)、杨梅素(4)、杨梅素-3-O-α-L-鼠李糖苷(5)、没食子酸(6)、熊果酸(7)和1,3-二羟基-2-(2′-甲基丙酰基)-5-甲氧基-6-甲基苯(8)。其中，化合物1为新黄酮醇苷类化合物，化合物2～7为首次从该植物中分离得到，化合物8为首次从植物中分离得到。     

北沙参中的新8-O-4′型异木脂素

为了研究北沙参的化学成分，通过大孔树脂吸附柱色谱、 Sephadex LH-20及反相ODS柱色谱进行化合物的分离， 利用多种波谱技术鉴定化合物结构。从醇提物的乙酸乙酯溶部分， 分离得到1个新化合物， 命名为可来灵素D(glehlinoside D，1)， 以及5个已知化合物： 裂异落叶松脂醇(2)、 阿魏酸(3)、 咖啡酸(4)、 香草酸(5)和丁香苷(6)。化合物1为一新的8-O-4′型异木脂素类化合物。