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AIM:To observe the therapeutic effect of stachydrine hydrochloride on experimental acute cerebral infarction in rats and to explore the underlying mechanisms .METHODS:SD rats ( n=75) were randomly divided into 5 groups:sham group, cerebral infarction model group , and stachydrine hydrochloride (10 mg/kg, 20 mg/kg and 40 mg/kg) treatment groups .After the establishment of cerebral infarction model , the rats were given stachydrine hydrochloride at dose of 10 mg/kg, 20 mg/kg or 40 mg/kg by gavage daily for 14 d.The impairment of neurological function in each group was scored .The cerebral infarction volume and brain water content were measured .Moreover , the protein levels of β-cate-nin, cyclin D1, glycogen synthase kinase 3β(GSK-3β) and p-GSK-3βin the brain tissues were detected by Western blot . RESULTS:Compared with cerebral infarction group , the score of neurological function impairment , cerebral infarction volume and brain water content were significantly decreased in stachydrine hydrochloride treatment groups .In addition , the protein levels of β-catenin, cyclin D1 and p-GSK-3βwere markedly increased after stachydrine hydrochloride treatment . CONCLUSION:Stachydrine hydrochloride protects against experimental acute cerebral infarction through activation of Wnt/β-catenin signaling pathway .
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AIM:To observe the therapeutic effect of stachydrine hydrochloride on experimental acute cerebral infarction in rats and to explore the underlying mechanisms .METHODS:SD rats ( n=75) were randomly divided into 5 groups:sham group, cerebral infarction model group , and stachydrine hydrochloride (10 mg/kg, 20 mg/kg and 40 mg/kg) treatment groups .After the establishment of cerebral infarction model , the rats were given stachydrine hydrochloride at dose of 10 mg/kg, 20 mg/kg or 40 mg/kg by gavage daily for 14 d.The impairment of neurological function in each group was scored .The cerebral infarction volume and brain water content were measured .Moreover , the protein levels of β-cate-nin, cyclin D1, glycogen synthase kinase 3β(GSK-3β) and p-GSK-3βin the brain tissues were detected by Western blot . RESULTS:Compared with cerebral infarction group , the score of neurological function impairment , cerebral infarction volume and brain water content were significantly decreased in stachydrine hydrochloride treatment groups .In addition , the protein levels of β-catenin, cyclin D1 and p-GSK-3βwere markedly increased after stachydrine hydrochloride treatment . CONCLUSION:Stachydrine hydrochloride protects against experimental acute cerebral infarction through activation of Wnt/β-catenin signaling pathway .
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To build an evaluation standard for quality of Leonuri Herba standard decoction. 13 batches of Leonuri Herba standard decoction with different quality were prepared. The contents of leonurine hydrochloride and stachydrine hydrochloride were determined; then the transfer rate and the extract rate were calculated and pH value was measured; and HPLC fingerprint method was established for analysis. The results of 13 batches of samples revealed that the transfer rate of leonurine hydrochloride and stachydrine hydrochloride was 30.0%-53.4% and 67.0%-82.6%, respectively; the extract rate was 12.1%-18.3% and the pH value was 5.87-6.22. Moreover, 12 common chromatographic peaks were determined based on fingerprint by using Similarity Evaluation System for Chromatographic fingerprint of Traditional Chinese Medicine (2012A). The similarity of 13 batches of samples was analyzed and compared, and the results showed that the similarity was higher than 0.9. In this study, the preparation method for Leonuri Herba decoction was standard, with high similarity in fingerprint, showing high precision, stability and repeatability in fingerprint analysis. Thus, this study can provide a reference for the quality control of Leonuri Herba dispensing granules.
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To optimize the concentrate process of alkaloid from Leonurus japonicus by nanofiltration-ultrafiltration coupling technology with response surface methodology. The experiment showed that after ultrafiltration pre-treatment, the total protein removal rate was 94.38% in aqueous extract from L. japonicus, and the nanofiltration technology had obvious advantages over the conventional concentrate process. The optimal concentrate conditions were as follows：molecular weight cut-off 450, pH 3.07, concentration of stachydrine hydrochloride 80.15 mg•L⁻¹, and concentration of the total alkaloid 285.73 mg•L⁻¹. The cut-off rate was 93.37% and 95.85% respectively for stachydrine hydrochloride and the total alkaloid under the optimum conditions, with a relative error of 0.79% and 1.16% respectively. The combination of Box-Behnken design and response surface analysis can well optimize the concentrate process of L. japonicus by nanofiltration, and the results provide the basis for nanofiltration concentrate for heat-sensitive traditional Chinese medicine.
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Objective To study the difference ofLeonurus japonicus germplasm resources and provide good materials for breeding.Methods Totally 20 wildLeonurus japonicus germplasm resources from different places of China were collected. Experiment was conducted in homogeneous garden in Quzhou City of Zhejiang Province. Phenological phase, cold endurance and habitat adaptability were observed; the plant height, fresh weight and dry weight in early flowering were measured; the contents of stachydrine hydrochloride and leonurine hydrochloride were determined in early flowering and out-of-season cultivation.Results Considering the habitat adaptability, dry plant weight in early flowering, the contents of stachydrine hydrochloride and leonurine hydrochloride separately in early flowering and out-of-season cultivation, it was believed that the germplasms from Linbao County, Sheqi County in Henan Province and Guidong County in Hunan Province were better, in which Linbao germplasm was the best: the dry plant weight was 30.6 g, the content of stachydrine hydrochloride and leonurine hydrochloride were 1.31% and 0.19% respectively in early flowering, and were 3.44% and 0.37% respectively under anti-season cultivation, and it can be well adapted in Zhejiang Province.Conclusion The germplasm ofLeonurus japonicas from Lingbao can be the best materials for breeding.
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To establish the quantitative method of stachydrine hydrochloride and leonurine hydrochloride in the preparations of Leonuri Herba. The contents of stachydrine hydrochloride and leonurine hydrochloride in the preparations of Leonuri Herba were determined by HPLC-MS. The chromatographic column was Waters XBridge Amide(4.6 mm×250 mm,5 μm). The mobile phase was acetonitrile-0.1% formic acid in gradient mode,at the flow rate of 1.0 mL• min⁻¹,with the split ratio of 1∶4. MS conditions for the ESI ion source,positive ion mode,selective ion scan(SIM) of stachydrine hydrochloride(m/z 144.0) and leonurine hydrochloride(m/z 312.0) was measured. The linear ranges of stachydrine hydrochloride was 0.562 8-281.4 μg•L-1（r=0.999 8）. The linear ranges of leonurine hydrochloride was 0.521 2-260.6 μg•L-1（r=0.999 8）. The method is accurate,simple,and reliable,and can be used to determine the contents of stachydrine hydrochloride and leonurine hydrochloride in the preparations of Leonuri Herba.
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Objective To control the quality of Herba Leonuri by studying the content of stachydrine hydrochloride in different parts of it. Method A HPLC method was developed for the content detetmination of stachydrine hydrochloride in flower, stem, leafage from Herba Leonuri. Spherisorb SCX colum was used with mobile phase of 20 mmol/L sodium dihydrogen phosphate (containing 0.04% triethylamine and 0.15% phosphoric acid). The colum temperature was 25 ℃, the detective wavelength was 192 nm. Results Stachydrine hydrochloride content was the highest in leafage and the lowest in stem. Conclusion To ensure the quality of Herba Leonuri, it is significant to choose medical material with more leafage.
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OBJECTIVE:To establish TLC scanner method for the determination of stachydrine hydrochloride in xue?niaokang granule.METHODS:Silica gel G thin layer plate was adopted in the determination with n-butanol-hydrochloric acid-ethyl acetate(8∶3∶1)used as developer and thin potassium heptaiodobismuthate test solution used as color-developing agent,the detection wavelength was515nm,the reference wavelength was700nm and the slit size was6.00mm?0.45mm.RESULTS:Good linear correlation with the peak area score of spots achieved when the concentration range of stachydrine hydrochloride was within5.06?g～25.32?g(r=0.9978),the average recovery was96.67%(RSD=2.01%).CONCLUSION:The method was accurate,simple,reliable,sensitive and reproducible,and it can be used for the determination of xueniaokang granule and its quality control.
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Objective To establish a HPLC method for the separation and determination of stachydrine hydrochloride in fresh medicinal material of Herba Leonuri.Methods The NH2 column(250 mm? 4.6 mm,5 ? m) was adopted with acetonitrile-water(80 ∶ 20) as mobile phase at a flow rate of 1.0 mL/min.The detection wavelength was at 192 nm.The column temperature was at 22 ℃.The solution for content determination was obtained after extracted with ethanol and then filtered and purified through an activated charcoal-aluminium oxide column with ethanol as eluant.Results Stachydrine hydrochloride in fresh Herba Leonuri can be separated and determined on NH2 column.A good linear range was within 2.29～ 11.46 ? g.The average recovery for stachydrine hydrochloride was 98.26 % and RSD was 1.52 %(n=5).Conclusion The method is steady and with good repeatability,and can be used to determine the content of stachydrine hydrochloride in fresh Herba Leonuri.
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Objective To establish the quality standard for Chanhou Zhuyu Capsules.Methods Leonurus Japonicus and Ligusticum chuanxiong in Chanhou Zhuyu Capsules were identified by TLC,and the content of stachydrine hydrochloride was determined by TLC Scanning.Results The quality identification by TLC was specific.Stachydrine hydrochloride had a good linearity in the range of 4.0～12.0 ?g,r=0.999 7,and the average recovery was 99.87 %,RSD=1.67 %.Conclusion The methods of identification and content determination are simple,reliable and reproducible.They can be used effectively for the quality control of Chanhou Zhuyu Capsules.
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Objective: To establish the quality standard for Guangxiao Liniao Capsules (Herba Verbenae, Herba Leonuri, Rhizoma Alismatis, Radix Linderae, etc.). Methods: Herba Verbenae, Herba Leonuri, Rhizoma Alismatis, Radix Linderae were identified by TLC, and the content of stachydrine hydrochloride was determined by TLC scanning. Results: Herba Verbenae, Herba Leonuri, Rhizoma Alismatis, Radix Linderae could be identified by TLC. Stachydrine hydrochloride showed a good linear relationship at a range of 4?g～20?g, r = 0.9975. The average recovery was 97.9%, and RSD was 1.20%. Conclusion: The methods are accurate and can be used for the quality control of Guangxiao Liniao Capsules
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Objective:To investigate the quality of Herba Leonuri Preparations sold in market. Methods: The alkaloid content in preparations was determined according to the determination method in China Pharmacopoeia (1995). Results: There were significant differences among Herba Leonuri Preparations from different pharmaceutical factories, place of origin and batches. Conclusions: It is suggested that the determination item of stachydrine hydrochloride be added.