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Objective: In this study, InDel markers were developed based on the high frequency Insertion/Deletion region of chloroplast genome of Ligusticum. Germplasm identification and phylogenetic development of Ligusticum chuanxiong and its common adulterants were studied with universal barcode. Methods: The 26 samples of L. chuanxiong and its common adulterants were amplified and sequenced by eight DNA universal barcodes: ycf1, matK, ITS2, rpoC1, rbcL, rpoB, trnK, and psbA-trnH. Genetic distance statistics, barcoding gap and phylogenetic tree analysis methods were used to study the phylogenetic relationship and phylogeny of L. chuanxiong. At the same time, the evolutionary tree was constructed to study molecular identification of L. chuanxiong and its common adulterants. Results: The results showed that rbcL conserved site was the highest (97.32%) with the highest GC content (44.9%). The rbcL+rpoB fragment had the smallest average intraspecific genetic distance (0.002 5). The psbA-trnH sequence fragment had the largest average interspecific genetic distance (0.429 2). The trnK and rbcL+rpoB sequence had the highest interspecific genetic distance. The overlap of the "barcoding gap" region of psbA-trnH was the least. The species of L. chuanxiong and other adulterated species were not accurately identified by the eight pairs of DNA barcodes. The cluster analysis of 24 InDel markers could accurately identify genuine L. chuanxiong and classify the species of L. chuanxiong and its adulterants into four categories, one of which was genuine L. chuanxiong collected from Sichuan. Conclusion: The ability of InDel markers to identify authentic L. chuanxiong and its common adulterants was higher than that of common barcode. According to the above studies, it is found that it is impossible to distinguish L. chuanxiong and its common adulterants by the traditional DNA barcodes because of the large difference in genetic components. The newly developed InDel molecular markers can effectively identify L. chuanxiong and its commonly used adulterants, and provide an effective method for the genuineness of L. chuanxiong at molecular level.
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Objective: Owing to fibrinogen dosage, this study is to develop a method for the bioassay of anti-gel bioactivity of Chuanxiong Rhizoma in vitro, and evaluate the antithrombotic effect of Chuanxiong Rhizoma and its related Chinese patent medicines. Methods: Chuanxiong Rhizoma powder was extracted in ethanol and water, respectively. The mixed extract was used as sample to quantify the level of fibrinogen when the stable fibrin gel was formed in vitro. The fibrinogen dosage was chosen as biomarker for anti-gel bioactivity. Sodium ferulate was chosen as reference. The anti-gel bioactivity of Chuanxiong Rhizoma was calculated according to the direct measurement belong to the bioassay statistical method in the Chinese Pharmacopoeia 2015 (Vol. 4). Moreover, the amounts of anti-gel bioactivity were quantified in nine Chuanxiong Rhizoma samples including Chuanxiong Rhizoma raw materials, decoction pieces, and related Chinese patent medicines to evaluate the applicability for this developed method. Results: Both sodium ferulate and the extract of Chuanxiong Rhizoma showed significant anti-gel bioactivity (P < 0.01). The RSD for the amounts of anti-gel bioactivity was 4.00% (n = 6) in six replicated tests with the confidence limit rate of 7.82% (n = 6). The amounts of anti-gel bioactivity were significant difference among the various types of Chuanxiong Rhizoma samples, i.e. 0.72-1.14 U/g for five Chuanxiong Rhizoma raw materials, 0.68 and 1.32 U/g for Chuanxiong Rhizoma decoction pieces and processed slice with yellow wine, 2.56 and 2.51 U/g for Tongmai Granules and Xuefu Zhuyu Tablets. Conclusion: The developed method can accurately quantify the level of anti-gel bioactivity in Chuanxiong Rhizoma raw materials, decoction pieces and related Chinese patent medicines to assess their antithrombotic effect.
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Objective: To study the phthalides with vasodilating activity of alcohol extracts from stems and leaves of Ligusticum chuanxiong. Methods: Compounds were isolated by extensive chromatographic techniques including MCI, silica gel, Sephadex LH-20, ODS, and semi-preparative HPLC. The vasodilating effect of phthalide and dimer phthalide on thoracic aortic ring in vitro was investigated. Results: Ten phthalides were isolated and identified, and their structures were elucidated as Z-3-butylidenephthalide (1), senkyunolide-E (2), Z-senkyunolide-H (3), (3Z,7β)-3-butenyl-7-hydroxy-4,5,6,7-tetrahydrophthalide (4), neoligustilide (5); Z,Z’-3.3’a,7.7’a-diligustilide (6), 3Z,3Z’-6,8’,7,3’-diligustilide (7), Z-tokinolide A (8), (3’Z)-(3S,8R,3a’S,6’R)-4,5-dehydro-3.3’a,8.6’-diligustilide (9), and (3Z)-(3aR,6S,3’R,8’S)-3a.8’,6.3’-diligustilide (10). Compound 1, 6-9 significantly reduced the strain of thoracic aortic ring in KCl preconstricted rats. The diastolic rate of compounds 6-8 were 60%, 52% and 70% at the highest concentrations (12 μmol/L), respectively. The EC50 was 9.46, 11.86 and 8.73 μmol/L, respectively. Firstly isolated from the stems and leaves of L. chuanxiong. Compounds 6, 8 are firstly isolated from L. chuanxiong. The test compounds could dilate blood vessels, and the activity of dimer phthalide was better than that of phthalide.
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Objective: To analyze the rules of coronary heart disease syndrome differentiation based on the data mining technology. Methods: First of all, the famous medical records and prescriptions were collected and normalized to construct the database of coronary heart disease prescriptions. Then, IBM SPSS Statistics 20.0, a statistical software, was used to conduct statistics on the distribution of drug frequency, disease site syndrome frequency and disease syndrome frequency. Finally, the Apriori algorithm was applied to carry out data mining and analyze the underlying law of drug compatibility. Results: A total of 145 prescriptions were selected and analyzed, including 216 Chinese medicines, eight syndromes of disease location and 12 syndromes of disease nature. Forty-six herbs with higher frequency were found, and the top five TCM herbs were Salvia miltiorrhiza, Glycyrrhiza uralensis, Trichosanthes kirilowii, Pinellia ternata and Ligusticum chuanxiong. The main syndromes of disease location were heart, kidney, spleen and liver, while the main syndromes of disease nature were blood stasis, qi deficiency, phlegm, deficiency of yang, qi stagnation, and deficiency of yin. When the minimum support was 15% and the minimum confidence coefficient was 70%, the association rule analysis results showed that a total of 15 association rules for two-herb-combinations and 26 association rules for the trigeminy medications were obtained. The most frequency two-herb-combinations were Allium macrostemon→T. kirilowii", "Schisandra chinensis→Ophiopogon japonicus", "Curcumae Radix→S. miltiorrhiza. The most frequently used trigeminy medications were A. macrostemon + P. ternata→T. kirilowii, S. miltiorrhiza + A. macrostemon→T. kirilowii and A. macrostemon + G. uralensis→T. kirilowii. Conclusion: The syndrome differentiation and medication of TCM in the treatment of coronary heart disease mainly focus on invigorating the circulation of blood and resolving stasis, moving qi, nourishing and replenishing qi, clearing up phlegms. It is consistent with the main syndromes of the disease nature, such as blood stasis and qi deficiency, and the herbs mostly return to the heart meridian, which is consistent with the main syndromes of the disease location. The rules of syndrome differentiation and medication of based on the data mining technique have great value for clinical medication guidance and application to analyze the prescription for coronary heart disease.
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OBJECTIVE: To investigate and apply the bioaccessibility of Cd in traditional Chinese medicines (TCMs) to human health risk assessment, and provide reference for the formulation of limit standards. METHODS: The contents of Cd in Coptis chinensis Franch., Ligusticum chuanxiong Hort., Lindera aggregate (Sims) Kosterm., and Curcuma kwangsiensis S.G. Lee et C.F. Liang were determined by inductively coupled plasma mass spectrometry (ICP-MS). The bioaccessibility of Cd in TCMs was investigated by in vitro physiologically based extraction test(PBET) digestion method. And the target hazard quotient (THQ) was applied to assess the health risks of Cd contents. RESULTS: The bioaccessibility of Cd for Coptis chinensis Franch., Ligusticum chuanxiong Hort., Lindera aggregate (Sims) Kosterm., and Curcuma kwangsiensis S.G. Lee et C.F. Liang was 49.0%, 40.0%, 44.9% and 65.7%, respectively. The results of risk assessment showed that before extraction, THQ values of Cd for Coptis chinensis Franch., Ligusticum chuanxiong Hort., Lindera aggregate (Sims) Kosterm., and Curcuma kwangsiensis S.G. Lee et C.F. Liang were 1.895, 1.045, 1.446 and 1.653, respectively; after extraction, THQ values of Cd for the four types of TCMs were 0.929, 0.418, 0.649 and 1.077, respectively. CONCLUSION: Cd in the four types of TCMs cannot be completely absorbed in the artificial gastrointestinal phase, and the risks of Cd are reduced after extraction. Except THQ for Curcuma kwangsiensis S.G. Lee et C.F. Liang, the THQ values are below the value of 1, indicating very low health risks for the other three types of TCMs after extraction. In this study, the bioaccessibility of heavy metals in TCMs is applied to the health risk assessment model in order to provide a scientific basis for objectively evaluating the health risks of heavy metals in TCMs and formulating a scientific and reasonable heavy metal limit standards.
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Objective To study the regulating effect of freeze-dried substances of Salvia miltiorrhiza and Ligusticum chuanxiong Decoction on lipid metabolism abnormality in focal cerebral ischemia rats. Methods The focal cerebral ischemia rat model was established by monofilament method. The rats were randomly divided into normal group, sham operation group, model group, and drug treatment group. Plasma was collected after the last dosage and UPLC-Q/TOF-MS was used to study the plasma lipidomics. The lipidomics data were analyzed by orthogonal partial least square discriminant analysis (OPLS-DA), and the lipid metabolites changes were examined before and after the intervention of S. miltiorrhiza and L. chuanxiong. Results The focal cerebral ischemia rat model was successfully repeated. The S. miltiorrhiza and L. chuanxiong Decoction freeze-dried substances obviously reversed the abnormal lipid metabolism profile in the focal cerebral ischemia rat model. The plasma lipid biomarkers of ischemia injury rat were PS (21:0/0:0), PG (12:0/17:0), C16 sphinganine, phytosphingosine, PE [18:1 (9Z)/0:0], PC (16:1/2:0), PC (0:0/18:0), PC (16:1/0:0), PC (16:0/0:0), PC (18:2/0:0), PC (18:1/0:0), PC (18:0/0:0), and PC (20:5/0:0), respectively. Conclusion S. miltiorrhiza and L. chuanxiong Decoction freeze-dried substances have protective effects on cerebral ischemia injury, which may be related to the regulation of abnormal lipid metabolism, especially for phosphatidylcholines (PCs).
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Objective: To observe the protective effect of Salvia miltiorrhizae and Ligusticum chuanxiong effective constituents: danshensu, protocatechuic aldehyde, ligustrazine, and ferulic acid combination on primary cultured hippocampal neurons injured by oxygen glucose deprivation, and find out an optimized combination. Methods Primary cultured rats hippocampal neurons waschosen as research objects by adopting immunohistochemistry of the neuron-specific enolase IgG to authenticate, then the OGD model of the hippocampal neurons injured by oxygen glucose deprivation was established. The non-cytotoxic dose range of danshensu, protocatechuic aldehyde, ligustrazine, ferulic acid, and nimodipine was studied by MTT method. The compatibility of components was arranged by L9 (34) orthogonal design. Primary cultured rats hippocampal neurons was divided into 12 groups: control group, model group, Nimodipine positive control group, and orthogonal design 1-9 group. The activity of LDH was measured by colorimetry, the activity of SOD was tested by WST-1 and the levels of MDA were examined by TBA. The levels of TNF-α, IL-1β, and IL-6 in cell culture supernate were examined by ELISA, the apoptosis of hippocampal neurons was detected by Fluorochrome Hoechst33258 staining and the cell early apoptosis rate was detected with flow cytometry. The results of orthogonal test was analyzed by using range analysis method. Results The orthogonal compatibility of reagents played significant roles against the hypoxia damage of hippocampal neurons, improved the cellular morphology obviously, reduced the activity of LDH, increased the activity of SOD and decreased the content of MDA significantly, inhibited the release of TNF-α, and reduced the content of IL-1β and IL-6, reduced the apoptosis of cells apparently. The effect of active ingredients of S. miltiorrhiza and L. Chuanxiong on LDH activity was danshensu > ligustrazine > protocatechuic aldehyde > ferulic acid, and the best combination was danshensu (120 μg/mL), protocatechuic aldehyde (120 μg/mL), ligustrazine (80 μg/mL), and ferulic acid (20 μg/mL). The effect on SOD activity was ferulic acid > ligustrazine > danshensu > protocatechuic aldehyde. The best combination was danshensu (120 μg/mL), protocatechuic aldehyde (120 μg/mL), ligustrazine (80 μg/mL), and ferulic acid (40 μg/mL). The order of influence on MDA content was danshensu > protocatechuic aldehyde > ferulic acid > ligustrazine, and the best combination was danshensu (60 μg/mL), protocatechuic aldehyde (60 μg/mL), ligustrazine (80 μg/mL), and ferulic acid (20 μg/mL). The effect on TNF-α content were: ligustrazine > protocatechuic aldehyde > danshensu > ferulic acid, and the best combination was danshensu (60 μg/mL), protocatechuic aldehyde (60 μg/mL), ligustrazine (40 μg/mL), and ferulic acid (10 μg/mL). The order of influence on the content of IL-1β was ligustrazine > ferulic acid > danshensu > protocatechuic aldehyde, and the best combination was danshensu (30 μg/mL), protocatechuic aldehyde (30 μg/mL), ligustrazine (80 μg/mL), and ferulic acid (20 μg/mL). The effect on the content of IL-6 was protocatechuic aldehyde > ligustrazine > ferulic acid > danshensu, and the best combination was danshensu (120 μg/mL), protocatechuic aldehyde (120 μg/mL), ligustrazine (80 μg/mL), and ferulic acid (10 μg/mL). The effect on the early apoptosis rate was ferulic acid > protocatechuic aldehyde > ligustrazine > danshensu, and the best combination was danshensu (60 μg/mL), protocatechuic aldehyde (30 μg/mL), ligustrazine (20 μg/mL), and ferulic acid (40 μg/mL). Conclusion The protective mechanism of the effective constituents of S. miltiorrhizae and L. chuanxiong were relied on reducing the oxidative damage, reducing inflammation damage, and antagonizing cell apoptosis. According to the experimental results, we need to change the prescription ratio and guide clinical medication for different clinical courses.
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Objective: To explore the effect of volatile oil of Ligusticum chuanxiong on vascular cognitive impairment (VCI) induced by lipopolysaccharide (LPS) and its related mechanism. Methods Male C57BL/C mice were randomly divided into sham-operated group, model group, memantine (10 mg/kg) group, and volatile oil of L. chuanxiong (30, 15, and 7.5 mg/kg) groups, drugs were given by ig administration once a day for 14 d. On day 8, LPS was injected into the lateral ventricle in mice. On day 11 and day 12, the cognitive ability were checked by the Y maze and step down test. On day 15 the mice were decapitated and the brain tissue was taken to determine the levels of monoamine oxidase (MAO), acetylcholinesterase (AchE), and acetylcholine (Ach); At the same time, the effect of the drug on superoxide dismutase (SOD) in mitochondria of brain in SD rats and the activity of Ec AchE, and BV-2 cells of microgliomas injured by LPS were also investigated in vitro. Results:LPS significantly increased the levels of MAO and AchE in the brain in mice, while decreased the levels of dopamine (DA), 5-hydroxytryptamine (5-HT), and norepinephrine (NE). The volatile oil of L. chuanxiong 30 or 15 mg/kg decreased the level of MAO and AchE in the brain of LPS mice. The volatile oil of L. Chuanxiong could significantly increase the times of spontaneous alternation reaction in Y maze in VCI mice, and the middle dose could significantly decrease the latency of VCI mice in step down test. In vitro, the volatile oil of L. chuanxiong 1 mg/mL had a weak inhibitory effect on AchE of electric eel. The volatile oil of L. chuanxiong 0.5 μg/mL could significantly increase the activity of SOD in brain mitochondria of rats, and decrease the levels of tumor necrosis factor-α (TNF-α) and nitric oxide (NO) increased significantly in BV-2 cells induced by LPS. Conclusion: The volatile oil of L. chuanxiong can significantly improve the cognitive ability of VCI induced by LPS in mice, and its mechanism may be related to inhibit the inflammatory response in the brain and alleviate the injury of neurons.
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Objective: To optimize the integration technology on field processing and processing crude drugs of Ligusticum chuanxiong (LC) and provide the scientific evidence for the integration of LC. Methods: Single factor test and orthogonal method was used to investigate the three significant influencing factors, which were water content, drying temperature, and slice thickness. Based on the appearance character and active component (ferulic acid, volatile oil, and extract) of LC, the integrated process was selected by comprehensive scoring method. To investigate the stability of the process, three batches of integrated slices were processed according to the optimized integrated process; And three batches of corresponding traditional slices were processed according to the method stipulated in Chinese Pharmacopoeia (2015 edition), and the quality of the two kinds of slices was compared. The rat migraine model was induced by nitroglycerin. The effects of two different processed slices on migraine were compared by recording the times of head flick and head scratching, the level of NO in serum and the activity of NOS in serum of rats. Results: The optimum integrated process was as follows: drying fresh LC until water content was 28%, cutting 2 mm thick slices, drying at 50 ℃ in drying box (about 6 h). The integrated process had good stability and repeatability, and the quality of integrated slices was close to that of traditional slices. Compared with the model group, two processed slices could significantly reduce the number of head flick and head scratch (P < 0.01, 0.001), and decreased the activity of NOS (P < 0.01), and the level of NO decreased significantly (P < 0.01). The pharmacodynamics of the two process pieces was similar. Conclusion: The integrated processing and traditional processing of Chuanxiong Rhizoma decoction has good similarity in character, effective component content and treatment of migraine. Moreover, the integrated processing has standardized the production of Chuanxiong Rhizoma decoction slices from the source. Under the premise of ensuring the quality of Chinese herbal slices, the production cycle is shortened and the production cost is saved. It is a kind of traditional Chinese medicine slices production mode which is worth popularizing.
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Objective: To study the chemical constituents of metabolites of Alternaria sp. IS275, an endophytic fungus isolated from Ligusticum chuanxiong. Methods: The extraction fraction of ethyl acetate from endophytic fungi fermentation broth of L. chuanxiong were isolated and purified by silica gel, Sephadex LH-20, reversed phase medium pressure chromatography, preparative TLC, and semi-preparative RP-HPLC. The structure of the separated chemical constituent was elucidated by modern spectroscopy technology. Results: A new lactone compound (+)-4,11-dihydroxy-2-dodecene-5-lactone was isolated from the ethyl acetate fraction of the metabolite of Alternaria sp. IS275, an endophytic fungus of L. chuanxiong. Conclusion: Compound 1 is a new lactone compound, named ligualterlide A.
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Objective: To investigate the anti-tumor effect of Ligusticum chuanxiong volatile oil in combination with temozolomide (TMZ) in the treatment of glioma, and illuminate its possible mechanism. Methods: After C6 glioma animal model was successfully finished, changes of body weight, survival status, tumor volume, and the tumor inhibition rate of rats were detected in the TMZ group and the compatibility group of the volatile oil of L. chuanxiong and TMZ. The concentration of TMZ in the internal and external fluids of U87-MG cells was measured by HPLC after the administration of TMZ and the combination of volatile oil of L. chuanxiong and TMZ, and the differences of the content were analyzed statistically. Western blotting was used to study the effects of TMZ monotherapy, L. chuanxiong volatile oil and TMZ on the expression of P-gp protein of U87-MG cells.Results: Compared with TMZ group, the tumor volume of rats in the compatibility group was significantly decreased (P < 0.05), which was positively correlated with the amount of volatile oil. At the same time, the weight gain of the rats in the compatibility group was increased and the survival status was better. Volatile oil of L. chuanxiong can promote the entry of TMZ into U87-MG cells, and with the increase concentration of volatile oil, the promoting effect was stronger. Volatile oil at 6.25 × 10-3 μL/mL can significantly increase the accumulation of TMZ in the internal fluid (P < 0.05). Western blotting analysis showed that P-gp expressions of cells treated with combination treatment with TMZ (20 μg/mL) and volatile oil [(3.125 × 10-3), (6.25 × 10-3) μL/mL] were markedly down-regulated (P < 0.01), when compared with TMZ group. Conclusion: The volatile oil of L. chuanxiong can promote TMZ entry into glioma cells and then enhance TMZ-induced anticancer efficiency in vivo by inhibiting P-gp expression.
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In recent years, Ligusticum chuanxiong Hort. is facing a serious international market challenge due to excessive levels of heavy metal cadmium. Reducing the cadmium content of L. chuanxiong has become an urgent problem to be solved. This paper first discusses the current research progress on high cadmium of L. chuanxiong including the sources of cadmium and the common measures to control the cadmium content of L. chuanxiong. It is pointed out that the existing control measures mainly focus on reducing the content of active cadmium in soil to reduce the toxicity of cadmium to L. chuanxiong. However, it has been proved that L. chuanxiong has an enrichment effect on soil cadmium, and the current research lacks to explore the reduction of active absorption of cadmium by L. chuanxiong. Based on this, this paper is the first time to analyze the feasibility of exogenous silicon to reduce the active absorption of cadmium by L. chuanxiong from a plant perspective, in order to provide a reference for exploring ways to effectively reduce the cadmium content of L. chuanxiong.
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Three new thionic compounds, ()-2-(2-carboxyl-2-hydroxyethylthio)-ferulic acid (), ()-2-methoxy-4-(3-(methylsulfonyl)prop-1-en-1-yl)phenol (), and thiosenkyunolide C (), together with two new aromatic glycosides ( and ) were isolated from the rhizome of Hort. Two known compounds ( and ) were also obtained. Their structures were elucidated based on extensive spectroscopic data (UV, IR, 1D and 2D NMR, and HR-ESI-MS). Furthermore the absolute configurations were established by comparison of their calculated and experimental circular dichroism spectra and by a dimolybdenum tetraacetate [Mo(AcO)]-induced circular dichroism procedure. All compounds were evaluated against lipopolysaccharide (LPS)-induced NO production in BV2 cells, and compounds and showed strong inhibitory activities with IC values of 2.03 and 3.09 µmol/L, respectively (positive control curcumin, IC = 6.17 µmol/L). In addition, compound showed weak proteintyrosine phosphatase-1B (PTP1B) inhibitory activity.
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Objective To investigate the chemical constituents of the rhizomes of Ligusticum chuanxiong and discuss the significance of first discovery of ginsenosides from the plant. Methods The compounds were isolated and repeatedly purified by column chromatographies such as macroperous resin, Sephadex LH-20, silica gel, and preparative TLC as well as semi-preparative RP-HPLC. Their structures were elucidated by physicochemical properties, NMR, and MS spectral analyses. Results Three ginsenoside compounds were isolated from the n-butanol extracts of rhizomes of L. chuanxiong, and their structures were identified as (20S)-ginsenoside Rh1 (1), (20R)-ginsenoside Rh1 (2), and (20R)-ginsenoside Rg3 (3). Conclusion Ginsenosides are isolated from the genus Ligusticum (Umbelliferae) for the first time, it is of great significance for clarifying pharmacodynamic material basis of the rhizomes of L. chuanxiong. These results also provide the reference data for further verifying the relevance of plant evolution and traditional efficacy between L. chuanxiong and Panax ginseng.
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Objective: To construct the three dimensional models of L. chuanxiong caffeic acid-3-O-methyltransferase (LCCOMT) and verify the model using site-directed mutagenesis technology. Methods: The three-dimensional model was constructed by homology modeling using the crystal structure of COMT from Medicago sativa as a temple. Caffeic acid was docked into the optimized model of LCCOMT to predict the active site. The predicted site was mutated using site-directed mutagenesis technology. Then, the activity of mutant enzyme was detected. Results: The molecular docking, which showed there were hydrogen bonds between His268 and 3-OH of caffeic acid, was successful. Two mutant enzymes, H268N and H268Q, lost 94.85% and 95.28% of their activity respectively. Conclusion: The His268 is confirmed as one of the key residues of LCCOMT. It may play a role as a base in the deprotonation reaction of the 3-OH of caffeic acid.
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Objective To optimize the extraction technology of ferulic acid (FA) in Ligusticum chuanxiong Hort. and tetrahydropalmatine in Corydalis yanhusuo from Kexintong dropping pill. Methods The extraction technology of L. chuanxiong was screened by the orthogonal test. The extration was refined with the high aperture jaffaite. The technology parameter for extracting C. yanhusuo by the classic method of acidic dissolution and alkaline precipitation was optimized by the orthogonal test. Results The more effective way to extract FA from L. chuanxiong was to extract with 12 times amount of 75% alcohol under reflux for 3.5 h. The chuanxiong extract of pH 3 at a flow rate of 5 ml/min on DM-130 macroporous resin column, was eluted with 80% ethanol. The eluent was collected and dried under vacuum. The more effective way to extract tetrahydropalmatine in C. yanhusuo was extracted with 22 times amount of acidic water (pH 4)for 3 h. Conclusion The optimum extraction is reasonable and practicable with high extraction ratio.
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Objective: To investigate the chemical constituents from Guanxinning Injection. Methods: The chemical constituents were repeatedly isolated by column chromatographic techniques including silica gel, ODS, Sephadex LH-20, and macroporous resin, and were purified by preparation HPLC chromatography and recrystallization, and their structures were identified by physicochemical properties and spectroscopic data. Results: Nineteen compounds were isolated and identified as protocatechuic aldehyde (1), vanillin (2), ferulic acid (3), 2-furoic acid (4), senkyunolide I (5), caffeic acid (6), salvianolic acid A (7), isosalvianolic acid A (8), salvianolic acid B (9), isosalvianolic acid C (10), rosmarinic acid (11), 1, 3-dicaffeoylquinic acid (12), senkyunolide H (13), 4-hydroxybenzoic acid (14), m-hydroxybenzoic acid (15), o-hydroxybenzoic acid (16), 4-hydroxycinnamic acid (17), methyl rosmarinate (18), and salvianolic acid N methyl ester (19). Conclusion: Nineteen compounds are isolated from the Guanxinning Injection for the first time, and all HPLC analyses and literature data show that compounds 2, 3, 5, 13-16 are originated from Ligusticum chuanxiong, compounds 7-11, and 18-19 from Salvia miltiorrhiza, and compounds 1, 4, 6, 12, and 17 from L. chuanxiong and S. miltiorrhiza.
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Objective: To optimize the extraction technology of ferulic acid (FA) in Ligusticum chuanxiong Hort. and tetrahydropalmatine in Corydalis yanhusuo from Kexintong dropping pill. Methods: The extraction technology of L. chuanxiong was screened by the orthogonal test. The extration was refined with the high aperture jaffaite. The technology parameter for extracting C.yanhusuo by the classic method of acidic dissolution and alkaline precipitation was optimized by the orthogonal test. Results: The more effective way to extract FA from L. chuanxiong was to extract with 12 times amount of 75% alcohol under reflux for 3.5 h. The chuanxiong extract of pH 3 at a flow rate of 5 ml/min on DM-130 macroporous resin column, was eluted with 80% ethanol. The eluent was collected and dried under vacuum. The more effective way to extract tetrahydropalmatine in C. yanhusuo was extracted with 22 times amount of acidic water (pH 4) for 3 h. Conclusion: The optimum extraction is reasonable and practicable with high extraction ratio.
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Objective:To establish a method for determining the concent of ferulic acid in compound Yangjiao soft capsules with HPLC.Method:To determine with HPLC using C18 column(250mm?4.6mm,5?m),methanol-1% acetic acid solution(37:63) as mobile phase.The flow rate was 1.0 ml/min and the detecting wavelength was 320nm.Results: Ferulic acid had good linear relation within the range from 6 to 60?g/min,r =0.9995.The average recovery rate was 100.3%,RSD=1.68%(n=9).Conclusion:The method is with good resolution,accurate and reproducible,which can be used for determining the content of ferulic acid in compound Yangjiao soft capsules.
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Object To provide a scientific basis for the identification and rational clinical use of the two herbal drugs. Methods Experiments were carried out by comparing their properties, microscopic characteristics, UV spectra, TLC and GC chromatograms. Results There are remarkable differences between the two crude drugs in these respects, and a convenient and effective method for their differentiation and identification has been established. Conclusion Ligusticum sinense Hort. and Ligusticum sinense Oliv. cv. chaxiong Mass should not be used one instead of the other.