Guided by anti-complementary activity, silica gel, Sephadex LH-20 and reversed-phase column chromatographies were used for fractionation and isolation of the ethyl acetate and n-butanol soluble fractions of Pogostemon cablin. Eighteen compounds were obtained, including 15 flavonoids: 5-hydroxy-3,7,3',4'-tetramethoxyflavone (1), 5-hydroxy-7,3',4'-trimethoxyflavanone (2), 5,4'-dihydroxy-3,7,3'-trimethoxyflavone (3), 5-hydroxy-3,7,4'-trimethoxyflavone (4), 5,4'-dihydroxy-7,3'-dimethoxyflavone (5), luteolin (6), quercetin-7,3', 4'-trimethyl ether (7), ermanine (8), 3,5,7- trihydroxy-3', 4'-dimethoxyflavone (9), quercetin (10), apigenin (11), kaempferol (12), 5-hydroxy-7,3',4'-trimethoxyflavone (13), kaempferol-7-O-beta-D-glucopyranoside (14) and kaempferol-3-O-beta-D-glucopyranoside-7-O-alpha-L-rhamnoside (15); one triterpenoid: oleanic acid (16); and 2 phenolic acids: vanillic acid (17) and benzylalcohol (18). The isolation of 5, 7, 8, 12-15 and 18 from the Pogostemon genus is reported for the first time. All isolates were evaluated for their in vitro anti-complementary activities on the classical pathway and alternative pathway. And the targets of the most potent constituent in complement activation cascade were identified using complement-depleted sera. Compounds 3, 7, 10, 12 and 16 exhibited anti-complementary activities toward the classical pathway and alternative pathway (CH50 0.072-1.08 g x L(-1), AP50 0.39-0.49 g x L(-1)), while 5 and 6 showed inhibitory effect on the classical pathway only. Mechanism study indicated that 7 interacted with C1q, C2, C5 and C9 components.
Complement Inactivating Agents ; chemistry ; isolation & purification ; pharmacology ; Lamiaceae ; chemistry

OBJECTIVETo study the anti-complementary phenolic acids from Lonicera japonica.

METHODThe anti-complementary activity-directed isolation was carried out with the hemolysis test as guide. All isolation was evaluated for their in vitro anti-complementary activities. The structures were identified by various spectroscopic data including ESI-MS, 1H-NMR, 13C-NMR data.

RESULTFourteen compounds were isolated from the EtOAc fraction of L. japonica extracts, including 8 phenolic acids: 5-O-caffeoylquinic acid (1), chlorogenic (2), 4-O-caffeoylquinic acid (3), 3,5-di-O-caffeoylquinic acid (4), 4,5-di-O-caffeoylquinic acid (5), 3,4-di-O-caffeoylquinic acid (6), caffeic acid (7) and methyl caffeate acid (8); 3 iridoids: secologanoside (9), sweroside (10) and secoxyloganin (11); and 3 flavonoids: luteolin (12), quercetin (13) and kaempferol (14). Compounds 1-9 and 11-14 showed anti-complementary activity in different extents and 3,5-di-O-caffeoylquinic acid (4) exhibited the most significant activity against the classical pathway.

CONCLUSIONCompound 14 is obtained from this plant for the first time, phenolic acids are the main anti-complementary constituents of L. japonica and 3,5-di-O-caffeoylquinic acid(4) is a potential complement inhibitor with strong activity, which worthy to be studied further in the future.

Complement Inactivating Agents ; chemistry ; isolation & purification ; pharmacology ; Hydroxybenzoates ; chemistry ; isolation & purification ; pharmacology ; Lonicera ; chemistry

OBJECTIVETo establish and optimize the preparation procedures of the anti-complementary polysaccharides from Houttuynia cordata.

METHODBased on the yield and anti-complementary activity in vitro, the conditions of extraction and alcohol precipitating process were optimized by orthogonal tests. The optimal condition of deproteinization was determined according to the results of protein removed and polysaccharide maintained. The best decoloring method was also optimized by orthogonal experimental design.

RESULTThe optimized preparation procedures were given as follows: extract the coarse powder 3 times with 50 times volume of water at 90 degrees C for 2 hours every time, combine the extracts and concentrate appropriately, equivalent to 0.12 g of H. cordata per milliliter. Add 4 times volume of 90% ethanol to the extract, allow to stand for 24 hours to precipitate totally, filter and the precipitate was successfully washed with anhydrous alcohol, acetone and anhydrous ether. Resolve the residue with water, add trichloroacetic acid (TCA) to a concentration of 20% to remove protein. Decoloration was at a concentration of 3% with activated carbon at pH 3.0, 50 degrees C for 50 min. The above procedures above were tested 3 times, resulting in the average yield of polysaccharides at 4.03% (RSD 0.96%), the average concentrations of polysaccharides and protein at 80.97% (RSD 1.5%) and 2.02% (RSD 2.3%), and average CH50 at 0.079 g x L-(-1) (RSD 3.6%).

CONCLUSIONThe established and optimized procedures are repeatable and reliable to prepare the anti-complementary polysaccharides with high quality and activity from H. cordata.

Complement Inactivating Agents ; isolation & purification ; pharmacology ; Houttuynia ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology

The polysaccharide B3-PS2 was extracted and purified from Herba Scutellariae Barbatae through chromatography of DEAE-cellulose and Sephacryl S-300 column. Average molecular weight of B3-PS2 was about 1,100 kD. It was composed of Glc, Gal and Ara in the ratio of 2.7:2.7:1.0, along with trace of Man, Rha, Fuc and Xyl. B3-PS2 inhibited complement activation on the classic pathways with CH50 value of (0.23 +/- 0.03) mg mL(-1). The targets of B3-PS2 upon the complement system were C1r, C1s, C3 and C4. These results suggested that anti-complementary activity of B3-PS2 was closed to its positive control heparin. It strongly suggested that the polysaccharide B3-PS2 from Herba Scutellariae Barbatae could be a potential candidate in treating those complement-associated diseases.
Complement Activation ; drug effects ; Complement Inactivating Agents ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; Polysaccharides ; chemistry ; isolation & purification ; pharmacology ; Scutellaria ; chemistry

OBJECTIVETo investigate the effects of complement inhibiting component of Ephedra sinica on immunological inflammation following acute spinal cord injury (SCI) in rats.

METHODSThe complement inhibiting component of Ephedra sinica was isolated by multiple precipitation steps and thin layer chromatography, and then the activity was analyzed. Fifty healthy SD rats were selected and randomly divided into the control group and the experimental group, 25 in each group. Induction of SCI was performed following a modified Allen's weight-drop method. The complement inhibiting component from Ephedra sinica (15 mg/kg) dissolving in 5 mL normal saline was immediately administered by gastrogavage after SCI, once daily. Equal volume of normal saline was administered to rats in the control group by gastrogavage. Hematoxylin and eosin (H&E) staining and C3 immunohistochemical staining were performed in SCI tissue at 12 h, day 1, 3, 7, and 14 after SCI. C3 positive expressions and myeloperoxidase (MPO) activity were assessed. Intercellular adhesion molecule-1 (ICAM-1) mRNA expression level was evaluated by Real-time PCR technique.

RESULTSC3 positive expression, MPO activity, and ICAM-1 mRNA level were significantly weaker in the Ephedra sinica group than in the control group at all time points (12 h, day 1, day 3, day 7, and day 14 after SCI) (P < 0.01, P < 0.05).

CONCLUSIONSThere existed complement system activation following acute SCI. The complement inhibiting component of Ephedra sinica significantly reduced immunological inflammation after SCI, and played an important role in secondary SCI.

Animals ; Complement Activation ; drug effects ; immunology ; Complement Inactivating Agents ; pharmacology ; Ephedra sinica ; chemistry ; Inflammation ; immunology ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; immunology ; metabolism ; pathology

AIM: To isolate and characterize the anti-complementary polysaccharide from the root of Bupleurum chinense. METHODS: Bioactivity-guided fractionation and purification was used to obtain the anti-complementary polysaccharide from the hot-water extract of the root of Bupleurum chinense. The polysaccharide was characterized by various chemical and spectral analyses. The anti-complementary activities were evaluated by hemolytic assay in vitro. The action targets were identified in the system with individual complement-depleted sera. RESULTS: A homogeneous polysaccharide BC-PS2 was isolated as an anti-complementary agent. It was identified as a branched polysaccharide with an average molecular weight about 2 000 KDa, composed of Glc, Ara, Gal, and Man in the ratio 3.5 : 2.4 : 2.0 : 1.0, respectively, along with a trace of Rha and Xyl, and only 1.11% of protein. The main linkages of the residues of BC-PS2 include terminal, 1, 6-linked, 1, 3-linked and 1, 3, 6-linked Glcp, terminal and 1, 5-linked Araf, terminal, 1, 4-linked, 1, 6-linked and 1, 4, 6-linked Galp, terminal, and, 1, 4-linked and 1, 4, 6-linked Manp. The bioassay experiments revealed that BC-PS2 inhibited complement activation on both the classical and alternative pathways, with CH50 and AP50 of (0.222 ± 0.013) and (0.356 ± 0.032) mg·mL(-1), respectively. Preliminary mechanism studies indicated that BC-PS2 interacted with C1q, C2, and C9 components. CONCLUSION The results demonstrated that BC-PS2 is an anti-complementary polysaccharide, and should be important constituent of the root of Bupleurum chinense for its application in the treatment of diseases associated with the excessive activation of complement system.
Adult ; Bupleurum ; chemistry ; Carbohydrate Sequence ; Complement Activation ; drug effects ; Complement Inactivating Agents ; chemistry ; isolation & purification ; pharmacology ; Hemolysis ; drug effects ; Humans ; Male ; Molecular Weight ; Plant Extracts ; chemistry ; isolation & purification ; pharmacology ; Plant Roots ; chemistry ; Polysaccharides ; chemistry ; isolation & purification ; pharmacology

To study the chemical constituents of Rabdosia japonica var. glaucocalyx and their anti-complementary activity on the basis of preliminary studies. Target isolation guided by anti-complementary activity test, compounds in the chloroform and n-butanol fractions were isolated and purified by silica gel and Sephadex LH-20 column chromatographies, and preparative HPLC. The structures were identified by various spectroscopic data including ESI-MS, 1H-NMR and 13C-NMR data. The compounds were evaluated for anti-complementary activity in vitro. Eleven compounds were isolated from the chloroform and n-butanol soluble fractions and identified as stigmasterol (1), stigmas-9 (11) -en-3-ol (2), glaucocalyxin D (3), kamebakaurin (4), maslinic acid (5), corosolic acid (6), minheryins I (7), diosmetin (8), caffeic acid ethylene ester (9), caffeic acid (10) and vitexin (11). Isoquercetrin, rutin, quercetin, 3-methylquercetin, luteolin, 7-methylluteolin, and apigenin which were isolated from the preliminary studies together with compounds 9 and 10 showed inhibition of the complement system by the classical pathway. Compounds 2, 4, 6-9 and 11 were obtained from this plant for the first time. Caffeic acid (10) showed the strongest activity in vitro with a CH50 value of 0.041 g x L(-1).
Animals ; Antioxidants ; pharmacology ; Caffeic Acids ; pharmacology ; Chromatography ; Chromatography, High Pressure Liquid ; Complement Hemolytic Activity Assay ; methods ; Complement Inactivating Agents ; chemistry ; pharmacology ; Cricetinae ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Erythrocytes ; drug effects ; Esters ; Ethylenes ; pharmacology ; Female ; Isodon ; chemistry ; Magnetic Resonance Spectroscopy ; Male ; Plant Components, Aerial ; chemistry ; Plant Growth Regulators ; pharmacology ; Sheep ; Spectrometry, Mass, Electrospray Ionization