The present study explored the physiological mechanism of the effects of different pH treatments on the growth, physiological characteristics, and stachydrine biosynthesis of Leonurus japonicus to provide references for the cultivation and quality control of L. japonicus. Under hydroponic conditions, different pH treatments(pH 5,6,7,8) were set up. The growth, physiology, and the content of stachydrine and total alkaloids of L. japonicus, as well as the content of key intermediate products in stachydrine biosynthesis pathway(i.e., pyruvic acid, α-ketoglutaric acid, glutamic acid, and ornithine) were monitored to explore the physiological mechanism of the effects of pH on the growth and active components of L. japonicus. The results showed that L. japonicus. could grow normally in the pH 5-8 solution. The pH treatment of neutral acidity was more conducive to the accumulation of photosynthetic pigments and the increase in soluble protein in leaves of L. japonicus. to promote its growth and yield. However, since stachydrine is a nitrogen-containing pyrrolidine alkaloid, its synthesis involves the two key rate-limiting steps of nitrogen addition: reductive ammoniation reaction and Schiff base formation reaction. High pH treatments promote the synthesis and accumulation of substrates and products of the above two reactions, indicating that the alkaline environment can promote the nitrogen addition reaction, thereby promoting the biosynthesis and accumulation of stachydrine.
Leonurus/chemistry* ; Hydroponics ; Alkaloids ; Nitrogen ; Hydrogen-Ion Concentration

OBJECTIVETo study the chemical constituents from the fruits of Leonurus japonicus.

METHODThe 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.

RESULTTwo 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).

CONCLUSIONCompound 1 is a new compound,named as leonujaponin A. Both compounds 1-2 are isolated for the first time from Leonurus plants.

Chromatography, High Pressure Liquid ; Fruit ; chemistry ; Leonurus ; chemistry ; Triterpenes ; chemistry ; isolation & purification

OBJECTIVETo study the enriching method of total flavonoid from Herba Leonuri with polyamide and macroporous resin.

METHODSeven 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.

RESULTD101 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%.

CONCLUSIONThe process is simple and convenient, and the regeneration of resin is easy, which has a good application foreground.

Absorption ; Flavonoids ; isolation & purification ; Leonurus ; chemistry ; Nylons ; chemistry ; Resins, Plant ; chemistry

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.
Magnetic Resonance Spectroscopy ; Leonurus/chemistry* ; Plants, Medicinal ; Diterpenes/chemistry* ; Plant Components, Aerial ; Molecular Structure

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.
Animals ; Drugs, Chinese Herbal ; adverse effects ; Humans ; Kidney ; drug effects ; pathology ; Leonurus ; chemistry

OBJECTIVETo develop an UPLC method for the determination of leonurine in traditional Chinese medicines.

METHODAn 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.

RESULTThe 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).

CONCLUSIONThe method is simple, reproducible and reliable. It can be used to control the quality of related drugs.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Gallic Acid ; analogs & derivatives ; analysis ; Leonurus ; chemistry

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.
Blood Platelets ; drug effects ; Coumarins ; chemistry ; pharmacology ; Leonurus ; chemistry ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; chemistry ; pharmacology

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.
Enzyme Inhibitors ; chemistry ; isolation & purification ; Fruit ; chemistry ; Glucosidases ; analysis ; antagonists & inhibitors ; Leonurus ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Extracts ; chemistry ; isolation & purification ; Terpenes ; chemistry ; isolation & purification

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.
Antineoplastic Agents ; chemistry ; isolation & purification ; pharmacology ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Humans ; K562 Cells ; Leonurus ; chemistry

OBJECTIVETo 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.

METHODSThe 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.

RESULTSThe 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.

CONCLUSIONThe 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.

DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; chemistry ; genetics ; Leonurus ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Sequence Analysis, DNA ; Species Specificity