New di-spirocyclic labdane diterpenoids from the aerial parts of Leonurus japonicus / 中国天然药物

Phytochemical investigation on the ethanol extract of a well-known medicinal herb Leonurus japonicus, led to the separation of 18 labdane type diterpenoids (1-18). Through comprehensive spectroscopic analyses and quantum chemical calculations, these compounds were structurally characterized as six new interesting 5,5,5-di-spirocyclic ones (1-6), two new (7 and 8) and six known (13-18) interesting 6,5,5-di-spirocyclic ones, a new rare 14,15-dinor derivative (9), and three new ones incorporating a γ-lactone unit (10-12). An in vitro neuroprotective assay in RSC96 cells revealed that compounds 7 and 12 exhibited neuroprotective activity in a concentration-dependent way, comparable to the reference drug N-acetylcysteine.

New diterpenoids isolated from Leonurus japonicus and their acetylcholinesterase inhibitory activity / 中国天然药物

Three new labdane diterpenoids, leojaponicone A (1), isoleojaponicone A (2) and methylisoleojaponicone A (3), were isolated from the herb of Leonurus japonicus. The chemical structures of these secondary metabolites were elucidated on the basis of 1D and 2D NMR, including HMQC, and HMBC spectroscopic techniques. All the new compounds were tested in vitro for their acetylcholinesterase and α-glucosidase inhibitory activity. Compounds 1-3 exhibited low inhibitory effects on α-glucosidase with respect to acarbose and exhibited high inhibitory effects on acetylcholinesterase with respect to huperzine A.

Leojaponic acids A and B, two new homologous terpenoids, isolated from Leonurus japonicus / 中国天然药物

The present study aimed at isolation and purification of the bioactive terpenoids from the herb of Leonurus japonicus by chromatographic separations such as silica gel, sephadex LH-20 and C18 reversed phase silica gel, as well as preparative HPLC. As a result, leojaponic acids A (1, C17H24O4) and B (2, C18H26O4), two homologous terpenoids, together with (-)-loliolide (3), 1-(3-ethylphenyl) ethane-1, 2-diol (4) and dibutyl phthalate (5), were isolated from the EtOH extract of L. japonicus. All the chemical structures of the isolates were elucidated on the basis of 1D and 2D NMR analyses. Compounds 1 and 2 were new terpenoids, and Compounds 3 and 4 were isolated and identified for the first time from this plant. In addition, the α-glucosidase and tyrosinase inhibitory activity of the new compounds were evaluated.

Application of Bioactive Natural Materials-based Products on Five Women's Diseases

Women's health has been threatened by various diseases mainly including heart disease, breast cancer, osteoporosis, depression, and autoimmune disease. But development of medication for these diseases has been restricted by high development costs and low success rates. Herein the attempt to develop valid bioactive materials from a traditional natural material has been made. Resveratrol has been reported to be effective in treatment of breast cancer and heart disease. Goji berry has received attention as a natural based therapeutic material to treat a diabetes, cardiovascular disease, and osteoporosis. Leonurus family has been reported to be effective particularly in pregnant women due to high contents of vitamin as well as stimulation of uterine contraction. Annona family has effects such as anti-anxiety, anticonvulsant and recently it is proposed to be as a therapeutic material to cure depression based on its strong antidepressant effect. Shiraia bambusicola has been utilized to cure angiogenesis-related disease from ancient China and furthermore recently it was proved to be effective in rheumatoid arthritis. Getting an understanding of utilization of these traditional natural materials not only enhances the interest in development of therapeutic materials for preventing and treating various women's diseases, but also makes it possible to develop novel therapeutic materials.

Agrobacterium-mediated transformation of LJAMP2 gene into 'Red Sun' kiwifruit and its molecular identification / 生物工程学报

Bacterial canker caused by Pseudomonas syringae pv. Actinidiae is one of the most important diseases of kiwifruit (Actinidia chinensis) and leads to considerable yield losses. In order to obtain transgenic plants with resistance for 'Red Sun' kiwifruit to canker disease, a non-specific lipid transfer protein-like antimicrobial protein gene (LJAMP2) from motherwort (Leonurus japonicus) was introduced into 'Red Sun' kiwifruit through Agrobacterium-mediated transformation. After two days of co-cultivation with A. tumefaciens strain LBA4404 harboring 35S:LJAMP2, the transformed explants were transferred to the selection medium containing 25 mg/L kanamycin+3.0 mg/L BA+1.0 mg/L NAA. The regeneration efficiency of kanamycin-resistant shoots reached to 85%. All (100%) of kanamycin-resistant shoots rooted on half-strength MS medium supplemented with 0.8 mg/L IBA and a total of 40 regenerated plantlets were obtained. PCR and histochemical GUS activity analysis show that 23 of 40 lines (57.50%) were positive, suggesting that the LJAMP2 gene was integrated into the genome of 'Red Sun' kiwifruit. Taken together, we established an efficient genetic transformation method for 'Red Sun' kiwifruit using A. tumefaciens and the transformation frequency reached 5.11%. This protocol will be useful for the genetic breeding of 'Red Sun' kiwifruit for improvement of disease resistance.

Coumarins from Leonurus japonicus and their anti-platelet aggregative activity / 中国中药杂志

Chemical constituents of Leonurus japonicus were isolated and purified by a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, MCI, and Rp C18. Structures of the isolates were determined by spectroscopic analysis as 10 coumarins: bergapten (1), xanthotoxin (2), isopimpinellin (3), isogosferal (4), imperatorin (5), meransin hydrate(6), isomeranzin(7), murrayone(8) , auraptenol(9), and osthol(10). In addition to compound 9, the others were isolated from the genus Leonurus for the first time. In the in vitro assay, compounds 4 and 8 significantly inhibited the abnormal increase of platelet aggregation induced by ADP.

Regulatory effect of leonurus extracts on hyperuricemia in rats / 中国中药杂志

In this study, SD rats were orally administrated with oteracil potassium (300 mg . kg-1 . d-1 ) to prepare the hyperuricemia model, and divided into normal, model, Allopurinol, LE high dosage, middle dosage and low dose (200, 100, 50 mg . kg-1 . d-1) groups. The rats were orally administrated with test drugs 1 hour later after being orally administrated with Oteracil potassium. After 7 days, serum uric acid, serum creatinine, uric acid and expression of relevant transporters in kidney were tested to study the regulatory effect of leonurus extracts on serum uric acid, renal function and relevant transporters in kidney of rats with hyperuricemia. Compared with the model group, the leonurus extract group could significantly down-regulate serum uric acid and creatinine levels of rats with hyperuricemia, and increase the urine uric acid level. Meanwhile, leonurus extracts could notably down-regulate the mRNA expressions of urate transporter 1 (URAT1) and glucose transporter 9 (GLUT9), up-regulate the mRNA expressions of organic cation transportanter (OCT) and Carnitine transporter (OCTN) and promote the excretion of uric acid of kidney.

A new C-28 nortriterpenoid from fruits of Leonurus japonicus / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents from the fruits of Leonurus japonicus.</p><p><b>METHOD</b>The compounds were extracted by 70% methanol, then isolated and purified by column chromatography on silica gel, RP-18, Sephedax LH-20 and HPLC. Their structures were determined on the basis of spectroscopic data and physicochemical property.</p><p><b>RESULT</b>Two 28-noroleanane-derived spirocyclic triterpenoids including a new acylated nortriterpenoid were isolated and identified as (17R)-19(18 --> 17)-abeo-2alpha,18beta,23-trihydroxy-3beta-E-feruloyoxy-28-norolean-12-ene (1) and phlomistetraol B (2).</p><p><b>CONCLUSION</b>Compound 1 is a new compound,named as leonujaponin A. Both compounds 1-2 are isolated for the first time from Leonurus plants.</p>

Metodologia para germinação de sementes de Leonurus sibiricus L / Methodology for Leonurus sibiricus L. seed germination

O objetivo do trabalho foi investigar a influência de diferentes temperaturas, o comportamento fotoblástico e a absorção de água de sementes de Leonurus sibiricus L. Essa espécie medicinal é originária da Índia, distribuída pela Ásia, África e América, utilizada no tratamento de reumatismo, problemas dermatológicos e respiratórios. Para tanto, as sementes foram submetidas a temperaturas entre 5 a 40ºC, com intervalos de 5ºC, e alternadas de 20/30, 20/25 e 25/30ºC, com 5 repetições de 50 sementes cada, em condições de luz e escuro. No estudo da absorção de água as sementes foram colocadas para germinar na temperatura de 20ºC e na presença da luz e pesadas para avaliar o ganho de água durante todo o processo germinativo, até a protrusão da radícula. Pelos resultados verificou-se que os maiores porcentuais de germinação e índice de velocidade de germinação ocorreram na temperatura constante de 20ºC, e nas temperaturas alternadas 20/25, 25/30, 25/30ºC sob luz. Houve germinação na temperatura mínima de 10ºC e na máxima 40ºC. No ensaio de absorção de água verificou-se que as sementes iniciam a protrusão da radícula com 65 horas de exposição e seguem padrão trifásico na curva de absorção. O modelo estatístico ajustado para a espécie foi y = 1,869 (1 - 0,414 exp ( -0,201t) + exp [-2,397 + 0,037 (t - 65)], com R²= 0,9998.

The aim of this study was to investigate the influence of different temperatures on the photoblastic response and water uptake of Leonurus sibiricus L. seeds. This medicinal species is from India and has been distributed over Asia, Africa and America, where it is used in the treatment of rheumatic, dermatological and respiratory disorders. Thus, seeds were subjected to temperatures between 5 and 40ºC, at 5ºC intervals, and 20/30, 20/25 and 25/30ºC alternate temperatures, with five replicates of 50 seeds each, under light and dark conditions. In the study of water uptake, seeds were allowed to germinate at 20ºC under light and weighed throughout the germinative process until radicle protrusion. The highest germination percentage and speed index were detected at 20ºC constant temperature and at 20/25, 25/30 and 25/30ºC alternate temperatures under light. There was germination at 10ºC (minimum temperature) and at 40ºC (maximum temperature). The study on water uptake showed that seeds began radicle protrusion at 65h exposure and follow the triphasic pattern in the uptake curve. The statistical model fit for the species was y = 1.869 (1 - 0.414 exp (-0.201 t) + exp [-2.397 + 0.037 (t - 65)], R² = 0.9998.

Advances in studies of renal adverse reactions of Leonurus heterophyllus / 中国中药杂志

Motherwort (Herb of Leonurus heterophyllus) was a traditional Chinese medicine used for the treatment of various kinds of gynaecological diseases, which was considered as non-toxic medicine since ancient times. However, adverse effects such as kidney damage, uterus damage, allergy and diarrhea were frequently reported recently. This paper reviews the possible target site, toxic dosage, chemical substance and other related factors of these kidney damage caused by motherwort from both the clinic and animal experiment view.

Genetic relationship analysis of different provenances of Leonurus japonicus by ISSR marker / 中国中药杂志

<p><b>OBJECTIVE</b>To study the genetic polymorphism and genetic relationship of different provenances of Leonurus japonicus.</p><p><b>METHOD</b>Genetic relationship in 19 provenances, which came from nationwide origins were measured by using polymerase chain reaction (PCR) with ISSR marker.</p><p><b>RESULT</b>Twenty ISSR primers were selected from 100 ISSR primers and used for ISSR amplification. A total of 164 bands were generated, of which 117 bands were polymorphic bands (the percentage of polymorphic band, PPB = 71.34%). The similarity coefficients were calculated with NTsys 2.10e software and the dendrogram were constructed with UPGMA. Nineteen samples were divided into 3 types using the cluster analysis according to their genetic similarity coefficient.</p><p><b>CONCLUSION</b>Results from the cluster analysis were correlated significantly with the morphological characteristics and geographical location of 19 samples. The data indicate that ISSR technique is useful to determine genetic diversity and genetic relationship among Leonurus provenances, providing a scientific basis for genetic breeding, differentiation and new cultivar selection.</p>

Determination of leonurine in leonurus granule and extractum by UPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To develop an UPLC method for the determination of leonurine in traditional Chinese medicines.</p><p><b>METHOD</b>An Acquity UPLC BEH C18 column (2.1 mm x 100 mm) with 1.7 microm particle size was used. The mobile phase was composed of methanol and ammonium formate buffer (pH 4.0) in gradient mode. The flow rate was 0.3 mL x min(-1) and the chromatograpic run time was 18 min for one sample.</p><p><b>RESULT</b>The results showed that there was significant difference in the content of leonurine in the leonurus products from different pharmaceutical companies. The leonurine content in those products is in the range of 45.6-193 microg x g(-1).</p><p><b>CONCLUSION</b>The method is simple, reproducible and reliable. It can be used to control the quality of related drugs.</p>

Chemical constituents and antitumor activity on leukemia K562 cell of Leonurus heterophyllus / 中国中药杂志

The chemical constituents of Leonurus heterophyllus were separated and purified by repeated column chromatography on silica gel, HPD 100, Sephadex LH-20, and PHPLC. Each compound was characterized by spectroscopic and physical data. Eight compounds have been purified and identified to be quercetin 3-O-robinobioside (1), rutin (2), isoquerci trin (3), hyperoside (4), quercetin (5), apigenin (6), genkwanin (7), and benzoic acid (8). Among them, compounds 2, 5-7 were isolated from L. heterophyllus for the first time; Compounds 1, 3, 4, 8 were obtained for the first time from the genus Leonurus. The in vitro activities against leukemia K562 Cells of pure components were evaluated by testing their IC50. Compounds 1-6, 8 exhibited in-vitro inhibitory activities against leukemia K562 cells in different extent.

Enriching of total flavonoids from Herba Leonuri with polyamide and macroporous resin / 中国中药杂志

<p><b>OBJECTIVE</b>To study the enriching method of total flavonoid from Herba Leonuri with polyamide and macroporous resin.</p><p><b>METHOD</b>Seven enriching and purifying methods were compared with the yield and purity as indexes. The method of enriching with polyamide and macroporous resin was confirmed and the process of purifying was determined by orthogonal design.</p><p><b>RESULT</b>D101 resin is packed by wet method, the ratio of diameter to height is 1:7. After mixed with the extract liquids, the weight of wet resin increased to 3 times of the dry resin. Evaporated the wet resin to dryness, mixed well with a little of 95% ethanol and dry polyamide powder, evaporated them to dryness again. Elute with deionized water until the effluent being colourless, then loaded it on the macroporous adsorptive resin, elute with 50% ethanol, the volume of effluents was collected to 7 times of the column volume. The purity of total flavonoids reached to 23%, while the diversion rate from raw Herba Leonuri was to 69%.</p><p><b>CONCLUSION</b>The process is simple and convenient, and the regeneration of resin is easy, which has a good application foreground.</p>

Experimental studies on compound Duzhong Jiangya prescription by semi-bionic extraction / 中国中药杂志

<p><b>OBJECTIVE</b>Optimize the technic condition of Duzhong Jiangya prescription with Semi-bionic Extraction.</p><p><b>METHOD</b>Using homogeneous design, under the same materials granularity, decoction temperature, consumption of water, filtration, concentration, and taking aucubin, hydrochloric acid stachydrine, maloicacid, baicalin, ethanol extract, dry extract as the indexes, the results were comprehensive considered to optimize the semi-bionic extraction condtions.</p><p><b>RESULT</b>The optimized SBE extraction conditions are the following: pH values of the water for the thrice extraction is 5.845 3, 7.496 1, 8.011 7, and the total extraction time is 3.418 0 h.</p><p><b>CONCLUSION</b>Considering the fact of manufacture, the SBE extraction conditions are the following: pH values of the water for the thrice extraction is 6.0, 7.5, 8.0, and the thrice extraction time is 1.5, 1.0, 1.0 h.</p>

Effect of Leonurus Heterophyllus Sweet on tissue factor transcription and expression in human umbilical vein endothelial cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of Leonurus Heterophyllus Sweet, (LHS) on tissue factor (TF) transcription and expression induced by thrombin in human umbilical vein endothelial cells (HUVECs).</p><p><b>METHODS</b>HUVECs were incubated with different concentrations of LHS and the TF mRNA expression was detected by reverse transcript-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>LHS treatment of HUVECs at different concentrations and for different times resulted in significant differences in TF expression (Plt;0.01). The transcription of TF in LHS-treated cells was significantly different from that of the blank control group (Plt;0.01).</p><p><b>CONCLUSION</b>LHS can decrease the expression of TF and intervene with TF transcription in HUVECs in vitro.</p>

Nuclear ribosomal DNA internal transcribed spacer 1 sequences of 4 Leonurus species / 南方医科大学学报

<p><b>OBJECTIVE</b>To analyze the nuclear ribosome DNA (nrDNA) internal transcribed spacer (ITS) sequences of 4 Leonurus species, and the possibility of using them for molecular authentication of the crude drugs from the genus.</p><p><b>METHODS</b>The nrDNA ITS sequence (including ITS1, 5.8S rDNA, ITS2, and partial 18S rDNA and 26S rDNA) of L. japonicus and its 3 adulterant species were amplified and sequenced, and CLUSTRAL X and MEGA software was employed for analysis.</p><p><b>RESULTS</b>The variation of ITS1 and ITS2 between L. japonicus and its adulterant species ranged between 7.2% and 18.8% and between 14.2% and 27%, respectively. The phylogenic tree derived from the dendrograms based on the ITS sequence data contained some discrepancy from the traditional classification.</p><p><b>CONCLUSION</b>The nrDNA ITS sequences can be used potentially as efficient markers for identification of L. japonicus and its adulterants, and further study is needed for studying the phylogeny of Leonurus.</p>

Experimental study of estrogenic activities of five kinds of Chinese herbal medicines / 中国中药杂志

<p><b>OBJECTIVE</b>To evaluate the estrogenic activity of several kinds of Chinese herbal medicines.</p><p><b>METHOD</b>Use zoopery and reporter gene technique to study the estrogenic activity of five Chinese herbal medicines. Zoopery: weanling female Kunming mice weight 9-12 g were administrated botanical extracts of Selaginella tamariscina, Pinus Massoniana, Corallodiscus flabellate, Dryopteris sublaeta and Leonurus heterophyllus, the positive control group with Nilestriol tablets and control group with water, respectiely. On the eighth day, the animals were sacrificed and the uteri were separated solely and weighed. Reporter gene technique: Induce the expression of reporter gene controlled by ERE and measure the activity of luciferase on cell's clear supernatant.</p><p><b>RESULT</b>The botanical extracts of S. tamariscina can increase weights of mice (P < 0.01); In the expression of reporter gene controlled by ERE, Either ERalpha or ERbeta's has estrogenic activity (P < 0.01). Follow in the zoopery we find the water part and the n-butanol part of S. tamariscina are the two active parts.</p><p><b>CONCLUSION</b>S. tamariscina and it's water part and n-butanol part have estrogenic activities, effect on ERbeta is greater than ERalpha.</p>

Probing into objectivity of adverse drug reaction of traditional Chinese medicine / 中国中药杂志

The adverse drug reaction (ADR) of traditional Chinese medicine (TCM) has been concerned nowadays. Some bans were published to limit their use abroad. There wasn't a feasible criterion to reply to the ADR of TCM yet in China. The ADR of 6 kinds of typical TCM which state attached importance to was chosen to discuss. Based on the retrospect investigation, the objectivity of ADR was explored in this study.