8-prenylnaringenin (8-PN) is a potent estrogen with high medicinal values. It also serves as an important precursor for many prenylated flavonoids. Microbial synthesis of 8-PN is mainly hindered by the low catalytic activity of prenyltransferases (PTS) and insufficient supply of precursors. In this work, a SfN8DT-1 from Sophora flavescens was used to improve the efficiency of (2S)-naringenin prenylation. The predicted structure of SfN8DT-1 showed that its main body is comprised of 9 α-helices and 8 loops, along with a long side chain formed by nearly 120 amino acids. SfN8DT-1 mutants with different side-chain truncated were tested in Saccharomyces cerevisiae. A mutant expressing the truncated enzyme at K62 site, designated as SfND8T-1-t62, produced the highest 8-PN titer. Molecular docking of SfN8DT-1-t62 with (2S)-naringenin and dimethylallyl diphosphate (DMAPP) showed that K185 was a potentially crucial residue. Alanine scanning within a range of 0.5 nm around these two substrates showed that the mutant K185A may decrease its affinity to substrates, which also indicated K185 was a potentially critical residue. Besides, the mutant K185W enhanced the affinity to ligands implied by the simulated saturation mutation, while the saturated mutation of K185 showed a great decrease in 8-PN production, indicating K185 is vital for the activity of SfN8DT-1. Subsequently, overexpressing the key genes of Mevalonate (MVA) pathway further improved the titer of 8-PN to 31.31 mg/L, which indicated that DMAPP supply is also a limiting factor for 8-PN synthesis. Finally, 44.92 mg/L of 8-PN was produced in a 5 L bioreactor after 120 h, which is the highest 8-PN titer reported to date.
Dimethylallyltranstransferase/metabolism* ; Flavonoids/metabolism* ; Molecular Docking Simulation ; Prenylation ; Saccharomyces cerevisiae/metabolism* ; Sophora/metabolism*

To investigate the effects of essential oil from three kinds of pungent herbs,namely Menthae Haplocalycis Herba,Atractylodis Rhizoma and Cnidii Fructus,on the transdermal absorption in vitro of alkaloids from Sophorae Flavescentis Radix. The modified vertical Franz diffusion cell was used to conduct a transdermal experiment in vitro with the isolated abdominal skin of the SD rats as the transdermal absorption barrier. The effects of such three kinds of pungent essential oil on percutaneous absorption of alkaloids from Sophorae Flavescentis Radix were investigated by determining the content of 6 alkaloids( oxymatrine,oxysophocarpine,N-methylcytisine,sophoridine,matrine,and sophocarpidine) in the transdermal acceptor with ultra-performance liquid chromatography-triple quadruple mass spectrometry( UPLC-TQ-MS) technique simultaneously. With enhancement ratio( ER) as the index,their effects on promoting penetration was as follows: 1% Atractylodis Rhizoma oil > 1% Cnidii Fructus oil > 3% Azone ≈ 3% Atractylodis Rhizoma oil > 5%Atractylodis Rhizoma oil > 3% Cnidii Fructus oil ≈ 5% Cnidii Fructus oil > 3% Menthae Haplocalycis Herba oil > 5% Menthae Haplocalycis Herba oil > 1% Menthae Haplocalycis Herba oil > Blank. The results showed that these three kinds of pungent essential oil could be used as enhancers for alkaloids of Sophorae Flavescentis Radix,providing scientific guidance for improving percutaneous absorption of alkaloids from Sophorae Flavescentis Radix.
Alkaloids ; metabolism ; Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; metabolism ; Oils, Volatile ; pharmacology ; Plant Roots ; chemistry ; Rats ; Rats, Sprague-Dawley ; Skin Absorption ; Sophora ; chemistry

To investigate the effect of flavonoids from Sophora flavescens in aging mice induced by D-galactose and its mechanism. Totally 60 mice were randomly divided into six groups: the control group, the model group, the piracetam group (positive control group) and flavonoids from S. flavescens low, medium and high doses groups. Except for the control group, all of the rest groups were subcutaneously injected with D-galactose (160 mg x kg(-1)) for successively 30 days to establish the sub-acute senescent model. Meanwhile, flavonoids from S. flavescens low, medium and high doses groups were respectively administered with 150, 300 and 600 mg xkg-('1)of flavonoids from S. flavescens for 30 days. The learning and memory abilities of mice were determined by avoiding darkness ex-eriment and jumping stair experiment. The contents of malondialdehyde (MDA) tumor necrosis factor-aα NF-aα the activities of superoxide dismutase (SOD) monoamine oxidase-B (MAO-B) Na'(+)K'(+)-ATPase and Ca2(+ )-ATPase in the brain of mice were deter-ined respectively after the behavioral experiments. The activity of lactic dehydrogenase ( DH) in blood serum was also determined and analyzed by microscope after HE staining to observe the changes in hippocampal organizational structure. Compared with the model group, flavonoids from S. favescens medium and high doses groups showed significantly increases in the latency of avoiding darkness and jumping stair experiments; flavonoids from S. fllvescens low, medium and high doses groups and the piracetam group showed de-reases in the numbers of errors in avoiding darkness experiment; the flavonoids from S. flavescens high dose group and the piracetam group showed reduction- n the number of errors in jumping stair experiment (P <0 . 5 or P <0 . 1). Flavonoids from S. flavescens me-ium and high doses groups and the piracetam group showed improvements in the activities of SOD, Na'(+)K'(+)ATPase in the brain of mice and declines in the contents of MDA and TNF-aα the activity of MAO-B in the brain of mice, the activity of LDH in blood serum (P <0 . 5 or P <0 . 1). Flavonoids from S. flavescens low, medium and high doses groups and the piracetam group also showed im-rovement in the activity of Ca2(+ )ATPase, with statistical difference from the model group (P <0 . 5 or P <0 . 1). The pathological result showed decreases in the number of cells of hippocampal dentate gyrus area, sparse cell arrangement, incomplete cellular mor-hology, scarce cytoplasm, blurred boundary between nucleus and cytoplasm, nuclei anachromasis, irregular pyknosis and unconspicu-us nucleoli in the model group. Compared with the model group, flavonoids from S. flavescens low, medium and high doses groups and the piracetam group showed improvements in hippocampal organization tissues. Flavonoids from S. favescens can improve the learning and memory ability of senescent mice induced by D-galactose. Its mechanism may be correlated with the enhancement of anti-oxidative actions by lowering TNF-aαcontent, which results in the stability of cell membrane and the reduction in MAO-B activity.
Aging ; drug effects ; metabolism ; psychology ; Animals ; Brain ; drug effects ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Flavonoids ; administration & dosage ; Galactose ; adverse effects ; Hippocampus ; drug effects ; metabolism ; Humans ; Learning ; drug effects ; Male ; Malondialdehyde ; metabolism ; Mice ; Sophora ; chemistry ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

BACKGROUNDHelicobacter pylori (H. pylori) infection could lead to most gastroduodenal diseases and is even identified as a carcinogen of gastric cancer. Total alkaloids of sophora alopecuroides (TASA) is widely used in herbal remedies to treat various infectious diseases, including stomach-associated diseases. This study is aimed at evaluating the antimicrobial activity of TASA on H. pylori-infected BALB/c mice mouse gastritis.

METHODSTotally 120 BALB/c mice were orally inoculated with H. pylori Bacterial liquid to construct BALB/c mice H. pylori infection gastritis animal model, after the model was successfully created. We randomly assigned 100 infected mice into 10 treatment groups, the first group (normal saline); the second group (bismuth pectin); the third group (omeprazole); the fourth group (TASA 2 mg/d); the fifth group (TASA 4 mg/d); the sixth group (TASA 5 mg/d); the seventh group (TASA + bismuth pectin); the eighth group (TASA + omeprazole); the ninth group (bismuth pectin + clarithromycin + metronidazole); the tenth group (omeprazole + clarithromycin + metronidazole), 5 other non-infected mice as negative control. Mice were orally inoculated twice a day and 7 days continuously. Then the mice were killed 4 weeks after treatment, we used realtime PCR to detect 16sDNA of H. pylori to test both the colonization and the clearance mice of bacteria of each treatment. We applied hematoxylin and eosin (HE) staining and immunostaining of mice gastric mucosa to observe the general inflammation and related factors interleukin 8 (IL-8), cyclooxygenase 2 (COX-2), and nuclear factor-kappa B (NF-κB) expression change after treatments.

RESULTSFirstly, we ensured that after 6-week intragastric administration, the bacteria colonization reached an exceed peak which is far higher than positive threshold (P < 0.001); secondly, after treatments, it is revealed that TASA combined with omeprazole or bismuth pectin showed promising antimicrobial activity against H. pylori as well as conventional triple therapy (P < 0.001); thirdly, HE staining showed that the inflammation on mice gastric mucosal membrane were also relieved obviously in TASA combined treatments and conventional triple therapy compared with normal saline treated mice, moreover, from immunohistochemistry results, H. pylori-induced IL-8, COX-2, and NF-κB were consistently suppressed in seventh, eighth, ninth, and tenth group to a certain extent.

CONCLUSIONThese results open the possibility of taking TASA as an anti-inflammatory agent for H. pylori gastritis.

Alkaloids ; pharmacology ; therapeutic use ; Animals ; Anti-Inflammatory Agents ; pharmacology ; therapeutic use ; Cyclooxygenase 2 ; metabolism ; Female ; Helicobacter Infections ; drug therapy ; Helicobacter pylori ; drug effects ; metabolism ; Immunohistochemistry ; Interleukin-8 ; metabolism ; Mice ; Mice, Inbred BALB C ; NF-kappa B ; metabolism ; Omeprazole ; therapeutic use ; Real-Time Polymerase Chain Reaction ; Sophora ; chemistry

The research aimed at investigating the physicochemical properties, stability and skin penetration in vitro of total alkaloids of Sophora flavescens nanoemulsion. Prepare total alkaloids of S. flavescens nanoemulsion and detect the determination of matrine and oxymatrine in the nanoemulsion using HPLC method. Transmission electron microscopy and laser particle size analyzer were utilized to detect the shape and size of the nanoemulsion respectively. And also the stability of nanoemulsion was studied under the conditions of low temperature (4 degrees C), normal temperature (25 degrees C) and high temperature (60 degrees C). Franz diffusion cell was used to research the transdermal absorption of nanoemulsion in vitro. The results found that the nanoemulsion we prepared presented appearance of rounded, uniform; its average diameter was (15.55 +/- 2.24) nm, and particle size distribution value was 0. 161; the appearance, diameter and percentage determination of total alkaloids of S. flavescens had no variations after 15 d under 4, 25, 60 degrees C respectively. The steady-state permeation rate was 4.564 1 microg x cm(-2) x h(-1), 24 h cumulative amount of penetration was 110.7 microg x cm(-2), which was 1.86 fold of 24 h cumulative amount of aqueous solution (59.41 microg x cm(-2)). All the results demonstrated total alkaloids of S. flavescens nanoemulsion had good permeability, and could provide a new preparation for its clinical application.
Alkaloids ; chemistry ; metabolism ; Animals ; Chemical Phenomena ; Drug Carriers ; chemistry ; Emulsions ; Male ; Nanostructures ; chemistry ; Rats ; Rats, Sprague-Dawley ; Skin Absorption ; Sophora ; chemistry

OBJECTIVETo investigate the anti-inflammatory efficacy accompanied by side effects of water extract and alcohol extract of Sophorae Tonkinensis Radix et Rhizome (STRR), their molecular mechanism, and interpret the relationship of "toxicity-effect" of toxic medicine.

METHODThe ear swelling by croton oil and granuloma by agar test models were used, water extract and alcohol extract of STRR of different dosages were administrated ig to mice to observe the assident toxicity, at the same time the activities of ALT, AST and the content of SOD, MDA,PEG2, NO, NOS, Cr, BUN, GSH, TG and Gn in serum were tested.

RESULTBoth water extract and alcohol extract of STRR have a strong inhibitory effect on ear swelling by croton oil and granuloma by agar. The activities of ALT, AST in serum were higher than that of normal group. SOD, MDA, PEG2, NO, NOS, GSH, TG and Gn had obvious changes.

CONCLUSIONBoth water extract and alcohol extract of STRR had an anti-inflammatory effect on acute and chronic inflammation. At the same time, side effects and liver toxicity. The anti-inflammatory effect of STRR in probable relation to the reduced inflammatory mediators release. Oxidative damnification might be one of the liver injury mechanism.

Alanine Transaminase ; blood ; metabolism ; Animals ; Anti-Inflammatory Agents ; administration & dosage ; adverse effects ; isolation & purification ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; isolation & purification ; Female ; Humans ; Inflammation ; drug therapy ; Kidney ; drug effects ; metabolism ; Liver ; drug effects ; enzymology ; metabolism ; Male ; Mice ; Rhizome ; chemistry ; Sophora ; chemistry

OBJECTIVETo investigate the protective effects of oxysophoridine (OSR) on primary cultured hippocampus neurons subjected to anoxia injury in neonatal rats and its mechanism.

METHODThe model of anoxia injury of hippocampus neurons in neonatal rats were primarily cultured in vitro by physical oxygen deficiency using glucose-free culture fluid. The survival rate of neurons, the leaking rate of lactate dehydrogenase (LDH), the intracellular contents of malondialdehyde (MDA) and nitric oxide (NO), the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and nitric oxide synthase (NOS) were measured. The intracellular free calcium concentration ([Ca2+]i) in hippocampus neurons were detected with Ca(2+)-sensitive dual wavelength fluorescence spectrophotometer.

RESULTNeuron death occurred in the anoxia injury model group with increase of LDH leaking rate, the contents of NO, MDA, intracellular [Ca2+] and the elevated activity of NOS while decreased activities of SOD and GSH-PX. The hippocampus neurons subjected to anoxia injury were alleviated in OSR (0.625, 5, 10 microg x L(-1)) group.

CONCLUSIONOSR has significant protective effects on hippocampus neurons subjected to anoxic injury. The mechanism of its protective effect may relate to its reduction of calcium overload and against oxidation injury.

Alkaloids ; administration & dosage ; Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Glutathione Peroxidase ; metabolism ; Hippocampus ; cytology ; drug effects ; enzymology ; metabolism ; Humans ; Hypoxia ; drug therapy ; enzymology ; metabolism ; prevention & control ; Malondialdehyde ; metabolism ; Neurons ; cytology ; drug effects ; enzymology ; metabolism ; Nitric Oxide Synthase ; metabolism ; Protective Agents ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Sophora ; chemistry ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the effect of total alkaloids of Sophora alopecuroides (TASA) on dextran sulfate sodium (DSS)-induced colitis in mice.

METHODSChronic experimental colitis was induced by administration of 4 cycles of 4% DSS. Fifty mice were randomly distributed into 4 groups (normal, DSS, DSS/high-dose TASA, and DSS/low-dose TASA groups) by a random number table with body weight stratification. Mice in the normal group (n=11) and DSS-induced colitis control group (n=15) received control treatment of 20 mL/kg distilled water; DSS plus TASA high- and low-dose groups (n=12 each) were treated with TASA solution (20 mL/kg) at the doses of 60 mg/kg and 30 mg/kg, respectively. The severity of colitis was assessed on the basis of clinical signs, colon length, and histology scores. Moreover, secretory immunoglobulin A (sIgA) and haptoglobin (HP) were analyzed by enzyme linked immunosorbent assay; intercellular adhesion molecule 1 (ICAM-1) and macrophage-migration inhibitory factor (MIF) gene expressions were analyzed by quantitative reverse transcriptase realtime polymerase chain reaction (qRT-PCR) using SYBA green I; and nuclear factor κ B (NF-κ B) expression and activation and p65 interaction with the promoter of ICAM-1 gene were assessed by Western blotting and chromatin immunoprecipitation assay.

RESULTSTASA administration significantly attenuated the damage and substantially reduced HP elevation and maintained the level of cecum sIgA. TASA inhibited the ICAM-1 gene expression and had no effect on MIF gene expression. Also, TASA was able to reduce phospho-I κ B α (p-I κ B α) protein expression; however, it had no effect on the activation of I κ B kinase α (IKK α) and inhibitor of NF-κ B α (I κ B α). Moreover, TASA inhibited the p65 recruitment to the ICAM-1 gene promoter.

CONCLUSIONSTASA had a protective effect on DSS-induced colitis. Such effect may be associated with its inhibition of NF-κ B activation and blockade of NF-κ B-regulated transcription activation of proinflammatory mediator gene.

Alkaloids ; pharmacology ; therapeutic use ; Animals ; Cecum ; drug effects ; metabolism ; pathology ; Colitis ; chemically induced ; drug therapy ; pathology ; prevention & control ; Colon ; pathology ; ultrastructure ; Dextran Sulfate ; Down-Regulation ; drug effects ; Female ; Haptoglobins ; metabolism ; I-kappa B Proteins ; metabolism ; Immunoglobulin A, Secretory ; metabolism ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Intestinal Mucosa ; drug effects ; pathology ; ultrastructure ; Macrophage Migration-Inhibitory Factors ; genetics ; metabolism ; Mice ; Mice, Inbred C57BL ; NF-KappaB Inhibitor alpha ; Phosphorylation ; drug effects ; Phytotherapy ; Promoter Regions, Genetic ; genetics ; Protective Agents ; pharmacology ; therapeutic use ; Protein Binding ; drug effects ; Signal Transduction ; drug effects ; Sophora ; chemistry ; Transcription Factor RelA ; metabolism

OBJECTIVETo study the hepatotoxicity mechanism of rats that induced by Sophorae Tonkinensis Radix et Rhizoma.

METHODThe SD rats were randomly divided into the control and Sophorae Tonkinensis Radix et Rhizoma (RRST) group, given distilled water and RRST 10 g x kg(-1) separately by orally for 7 days, and RRST 20 g x kg(-1) for other days until the 26th day. Blood was drawn from abdominal aorta after the rats were anesthetized by 25% urethane, and then centrifugally processed to get the serum for detection of ALT and AST. The liver tissues of control and experimental group were taken to prepare liver homogenate with cold NS, and centrifugally processed to get the supernatant. The activities of SOD and GSH, the content of gamma-GT and MDA were detected according to the methods of kit. The tumor necosis factor(TNF-alpha) was detected by ABC-ELISA. The expression of ICAM-1 was detected by immunohistochemistry.

RESULTAfter the rats were given RRST by oral for 26 days, the ALT and AST activities in serum increased, the content of GSH and the ratio of SOD and MDA decreased, the content of gamma-GT and TNF-alpha, the masculine expression of ICAM-1 increased.

CONCLUSIONAfter the rats were given RRST, the liver can be damaged obviously, and the mechanism of hepatotoxicity perhaps related to free radical and inflammatory factor.

Administration, Oral ; Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Drugs, Chinese Herbal ; chemistry ; toxicity ; Female ; Free Radicals ; metabolism ; Glutathione ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Liver ; drug effects ; enzymology ; metabolism ; Male ; Malondialdehyde ; metabolism ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rhizome ; chemistry ; Sophora ; chemistry ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; gamma-Glutamyltransferase ; metabolism

OBJECTIVETo study the modulatory effect of Sophora flaveacens and co-administration with Veratrum nigrum on the activity and mRNA expression of cytochrome P450 isoenzymes in rat liver.

METHODRat liver microsomal cytochrome P450, b5, aminopyrine N-demethylase (APND), p-nitrophenol-hydroxylase (pNPH) activities were quantitated by UV chromatography. The mRNA expression level of five CYP isoenzymes CYP1A1, CYP2B1/2, CYP2C11, CYP2E1 and CYP3A1 were detected by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTS. flaveacen and its combination with V. nigrum can dramatically reduce P450 and b5 protein content. Both single and combined use inhibited the activities of aminopyrine N-demethylase. At the mRNA level, the expression of CYP2C11 markedly induced exposure to V. nigrum, while decreased after combination with S. flaveacens. S. flaveacens can induce CYP2B1/2 gene expression, while the Sophora- Veratrum group showed weak inhibition. Both single and co-administrated group have some inhibitory effect on CYP3A1 gene expression, while CYP1A gene expression almost has no change in each group.

CONCLUSIONThese results indicated that compatibility of Sophora and Veratrum in a prescription also have herb-herb interactions based on P450. The possible mechanisms of its incompatible effects need to be further analysised via integrating with metabolic studies.

Animals ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Female ; Gene Expression Regulation, Enzymologic ; drug effects ; Liver ; drug effects ; enzymology ; Male ; Plant Extracts ; administration & dosage ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Sophora ; chemistry ; Veratrum ; chemistry