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OBJECTIVE To investigate the improvement effect and mechanism of calycosin （CA） on acute inflammatory injury secondary to intracerebral hemorrhage. METHODS Male C57BL/6 mice were injected with type Ⅶ collagenase into the basal ganglia to establish an intracerebral hemorrhage model， which were divided into sham-operation group（phosphate buffered saline instead of collagenase）， model group， and different CA dose groups（15，30，60，120 mg/kg）. Based on the modified neurological severity score （mNSS） to screen the intervention doses， the volume of intracerebral hemorrhage， brain water content， the expressions of ionized calcium-binding adaptor molecule 1 （Iba1） in brain tissue， Toll-like receptor 4 （TLR4） and its downstream inflammatory factors [tumor necrosis factor-α （TNF-α）， inducible nitric-oxide synthase （iNOS）， interleukin-1β （IL- 1β）] in brain tissue， and the apoptosis of cells in brain tissue were detected. Primary microglia were cultured in vitro， and the expressions of TLR4 and its downstream inflammatory factors were detected. Primary neurons and primary microglia were co- cultured in vitro， and the apoptosis of neurons was detected. RESULTS The doses of 30 mg/kg and 60 mg/kg were selected as intervention doses of CA for subsequent experiments. Compared with the sham-operation group， the mice in the model group had cerebral hemorrhage， the volume of cerebral hemorrhage and brain water content were significantly increased （P＜0.05）； the positive expression rate of Iba1 protein in brain tissue was significantly increased， and the relative expression levels of TLR4， TNF-α， IL-1β and iNOS protein in brain tissue were up-regulated significantly. The apoptosis rate also increased significantly （P＜0.05）. Compared with model group， the above indexes of the mice in the 30 and 60 mg/kg CA groups were significantly improved （P＜0.05）. CA significantlyreduced the relative expression levels of TLR4 and its downstream inflammatory factors in microglia， and reduced the apoptosis of neurons in the co-culture system of primary neurons and primary microglia （P＜0.05）. CONCLUSIONS CA can exert a protective effect on the brain， which may be related to relieving the secondary acute inflammatory injury after intracerebral hemorrhage by inhibiting TLR4-mediated inflammatory response.
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Objective:To establish a quality evaluation method for the simultaneous determination of Calycosin-7-O-β-D-Glucopyranoside and Lobetyolin in Danqi Xinmaikang boiled powders and pieces.Methods:Quantitative analysis of multi-components was performed to determine contents of Calycosin-7-O-β-D-Glucopyranoside and Lobetyolin with Calycosin-7-O-β-D-Glucopyranoside as the reference substance by single-maker (QAMS). The chromatogram conditions were established, with C18 column as solid phase, acetonitrile-water as flowing phase, 268 nm as detecting wavelength, 1.0 ml/min as flowing rate, 30 ℃ as column temperature, and 10 μl as injection volume.Results:The relative correction factor between Calycosin-7-O-β-D-Glucopyranoside and Lobetyolin was 1.14. There was no significant difference of measured values between the external standard method and QAMS ( P>0.05). With Calycosin-7-O-β-D-Glucopyranoside retention time of 1.00, the relative retention time of Lobetyolin was 1.51 and RSD was less than 5%. Conclusion:It is feasible and accurate to evaluate the quality of Danqi Xinmaikang boiled powders and pieces by QAMS.
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ObjectiveTo investigate the effect of calycosin-mediated glucoprotein130/Janus kinase/signal transducer and activator of transcription factor （GP130/JAK/STAT） signaling pathway on oxidative injury of astrocytes in spinal cord. MethodAstrocytes in rat spinal cord were isolated and identified by immunofluorescence detection of glial fibrillary acidic protein （GFAP）. The cells were respectively pre-treated with 5， 10， 20 μmol·L-1 calycosin for 12 h， and then 100 μmol·L-1 H2O2 （24 h） was added to induce oxidative injury. Cell counting kit-8 （CCK-8） assay was employed to detect cell proliferation and select the optimal concentration of calycosin. The following experimental groups were designed： control group， model group （100 μmol·L-1 H2O2）， calycosin group （20 μmol·L-1 calycosin）， calycosin + LY294002 group （20 μmol·L-1 calycosin + 10 μmol·L-1 LY294002）， and calycosin + Stattic group （20 μmol·L-1 calycosin + 3 μmol·L-1 Stattic）. CCK-8 assay and immunofluorescence method were used to detect the proliferation of cells and flow cytometry was applied to detect cell apoptosis and cycle. The protein expression of phosphorylated （p）-JAK2， p-STAT3， p-protein kinase B （Akt）， GP130， and interleukin-6 （IL-6） was detected by Western blotting. ResultCompared with the control group， the model group showed low proliferation activity and high apoptosis rate of cells （P<0.05）. Compared with the model group， calycosin （20 μmol·L-1） group displayed high proliferation activity and low apoptosis rate of cells （P<0.05）. Compared with calycosin （20 μmol·L-1） group， both phosphatidylinosirtol-3-kinases （PI3K） inhibitor LY294002 and STAT3 inhibitor Stattic significantly reduced the proliferation activity and increased the apoptosis rate of cells （P<0.05）. The protein expression of p-JAK2， p-STAT3， p-Akt， GP130， and IL-6 in the model group was higher than that in the control group （P<0.05）， and the expression of the above indicators was lower in each treatment group than in the model group （P<0.05）. ConclusionCalycosin can promote the proliferation and inhibit the apoptosis of astrocytes with oxidative injury by inhibiting the phosphorylation of PI3K/Akt pathway and JAK2/STAT3 pathway.
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ObjectiveTo investigate the effect of calycosin-mediated glucoprotein130/Janus kinase/signal transducer and activator of transcription factor （GP130/JAK/STAT） signaling pathway on oxidative injury of astrocytes in spinal cord. MethodAstrocytes in rat spinal cord were isolated and identified by immunofluorescence detection of glial fibrillary acidic protein （GFAP）. The cells were respectively pre-treated with 5， 10， 20 μmol·L-1 calycosin for 12 h， and then 100 μmol·L-1 H2O2 （24 h） was added to induce oxidative injury. Cell counting kit-8 （CCK-8） assay was employed to detect cell proliferation and select the optimal concentration of calycosin. The following experimental groups were designed： control group， model group （100 μmol·L-1 H2O2）， calycosin group （20 μmol·L-1 calycosin）， calycosin + LY294002 group （20 μmol·L-1 calycosin + 10 μmol·L-1 LY294002）， and calycosin + Stattic group （20 μmol·L-1 calycosin + 3 μmol·L-1 Stattic）. CCK-8 assay and immunofluorescence method were used to detect the proliferation of cells and flow cytometry was applied to detect cell apoptosis and cycle. The protein expression of phosphorylated （p）-JAK2， p-STAT3， p-protein kinase B （Akt）， GP130， and interleukin-6 （IL-6） was detected by Western blotting. ResultCompared with the control group， the model group showed low proliferation activity and high apoptosis rate of cells （P<0.05）. Compared with the model group， calycosin （20 μmol·L-1） group displayed high proliferation activity and low apoptosis rate of cells （P<0.05）. Compared with calycosin （20 μmol·L-1） group， both phosphatidylinosirtol-3-kinases （PI3K） inhibitor LY294002 and STAT3 inhibitor Stattic significantly reduced the proliferation activity and increased the apoptosis rate of cells （P<0.05）. The protein expression of p-JAK2， p-STAT3， p-Akt， GP130， and IL-6 in the model group was higher than that in the control group （P<0.05）， and the expression of the above indicators was lower in each treatment group than in the model group （P<0.05）. ConclusionCalycosin can promote the proliferation and inhibit the apoptosis of astrocytes with oxidative injury by inhibiting the phosphorylation of PI3K/Akt pathway and JAK2/STAT3 pathway.
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Aim To study the effects of calycosin on proliferation and migration of human triple-negative breast cancer MDA-MB-231 cells and the underlying mechanisms. Methods MDA-MB-231 cells were intervened by calycosin,and the proliferation ability was detected by CCK-8 method. The apoptosis and cycle of MDA-MB-231 cells were detected by flow cytometry. The effect of calycosin on the migration of MDA-MB-231 cells was observed using cell scratch. The mRNA expression of EMT related genes was detected by RT-PCR. The effects of calycosin on the expression of key proteins of Hippo pathway and EMT related proteins were detected by Western blot. Results Calycosin could significantly inhibit the proliferation,migration and apoptosis of MDA-MB-231 cells,and markedly inhibit the expression of Hippo signaling pathway and EMT related protein. Conclusion Calycosin may induce MDA-MB-231 cell apoptosis,block cell cycle and inhibit cell migration by inhibiting Hippo signaling pathway and EMT pathway.
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Astragali Radix, a medicinal herb for invigorating Qi, has anti-aging, anti-tumor, immunoregulatory, blood sugar-and lipid-lowering, anti-fibrosis, anti-radiation and other pharmacological effects. This article reviewed the studies about the chemical components and pharmacological effects of Astragali Radix. According to the theory of quality markers(Q-markers) of Chinese medicinal materials, we predicted the Q-markers of Astragali Radix from traditional efficacy, chemical component validity, measurability, plant phylogeny, and pharmacokinetis. The results showed that total polysaccharides, flavonoids(e.g., calycosin-7-O-β-D-glucoside, formononetin, calycosin, quercetin, and ononin), and saponins(e.g., astragalosides Ⅱ, Ⅲ, and Ⅳ) can be taken as the main Q-markers. This review lays a foundation for regulating the quality research and standard establishment of Astragali Radix, and benefits the control and quality supervision of the production process of Astragali Radix and its related products.
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Astragalus Plant , Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/pharmacology , Flavonoids , Plant RootsABSTRACT
The effects of water regulation on the biosynthesis of calycosin-7-O-β-D-glucoside in 2-year-old Astragalus membranaceus var. mongholicus were studied,and the mechanism was explained from the aspects of key enzyme gene expression and antioxidant enzyme system. The content of calycosin-7-O-β-D-glucoside was determined by HPLC,and the expression levels of six key enzyme genes( PAL,4 CL,CHS,CHI,IFS,13'H) in the synthesis pathway were analyzed by q RT-PCR. The activities of protective enzymes and contents of osmoregulation substances and malondialdehyde were also determined. In the water deficit group,the maximum concentration of calycosin-7-O-β-D-glucoside was 0. 49 mg·g-1 on the 24 th day of treatment. In the whole water regulation,the water deficit group outweighed the water adequate group in osmoregulation substance and MDA contents. The activities of A. membranaceus var.mongholicus antioxidant enzymes SOD,POD,and CAT increased during the initial period of water regulation,but decreased with time.The expression of PAL,CHS,and 13'H in the water deficit group was at a low level,and the 4 CL had active expression,slightly lower than that in the water adequate group. The expression of CHI and IFS elevated rapidly when water deficit occurred. Correlation analysis showed that the content of calycosin-7-O-β-D-glucoside was positively correlated with CHI expression( P<0. 01) and IFS expression( P<0. 05). Therefore,water regulation can change the accumulation pattern of calycosin-7-O-β-D-glucoside,and water deficit may be an effective way to increase its content. CHI and IFS are the key genes in response to water deficit.
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Astragalus propinquus/genetics , Biosynthetic Pathways , Glucosides , Isoflavones , WaterABSTRACT
Astragali Radix is one of the most commonly used medicinal materials. In recent years, its cultivated varieties and a variety of adulterants have flooded the market, which makes its quality uneven, and the development of quality control methods has become a research hotspot. Therefore, figuring out the quality markers of Astragali Radix is of great significance for its comprehensive evaluation. In this study, the fingerprints of 15 batches of Astragali Radix were established by HPLC, and the main components causing intergroup differences were screened out by PLS-DA. On the basis of literature review and network pharmacology analysis, the targets and pathways of active ingredients were obtained from SwissTargetPrediction, PubChem Compound and other databases, and then the "component-target-pathway" network was constructed with Cytoscape 3.7.1 for the prediction of potential quality markers. Twenty-eight common peaks were identified in the established fingerprint, and three differential components were selected as potential quality markers for Astragali Radix, which were astragaloside Ⅳ, calycosin-7-O-β-D-glucoside and ononin. The proposed method based on HPLC fingerprint of Astragali Radix is convenient and feasible, facilitating the improvement in its quality control.
Subject(s)
Astragalus Plant , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Plant Roots , Quality ControlABSTRACT
Objective: Osteoporosis has become the biggest cause of non-fatal health issue. Currently, the limitations of traditional anti-osteoporosis drugs such as long-term ill-effects and drug resistance, have raised concerns toward complementary and alternative therapies, particularly herbal medicines and their natural active compounds. Thus, this study aimed to provide an integrative analysis of active chemicals, drug targets and interacting pathways of the herbs for osteoporosis treatment. Methods: Here, we introduced a systematic pharmacology model, combining the absorption, distribution, metabolism, and excretion (ADME) screening model, drug targeting and network pharmacology, to probe into the therapeutic mechanisms of herbs in osteoporosis. Results: We obtained 86 natural compounds with favorable pharmacokinetic profiles and their 58 targets from seven osteoporosis-related herbs. Network analysis revealed that they probably synergistically work through multiple mechanisms, such as suppressing inflammatory response, maintaining bone metabolism or improving organism immunity, to benefit patients with osteoporosis. Furthermore, experimental results showed that all the five compounds (calycosin, asperosaponin VI, hederagenin, betulinic acid and luteolin) enhanced osteoblast proliferation and differentiation in vitro, which corroborated the validity of this system pharmacology approach. Notably, gentisin and aureusidin among the identified compounds were first predicted to be associated with osteoporosis. Conclusion: Herbs and their natural compounds, being characterized as the classical combination therapies, might be engaged in multiple mechanisms to coordinately improve the osteoporosis symptoms. This work may contribute to offer novel strategies and clues for the therapy and drug discovery of osteoporosis and other complex diseases.
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Objective To investigate the effect of calycosin on cerebral ischemia/reperfusion injury and its mechanism. Methods Forty SPF male SD rats were randomly divided into sham group, model group, calycosin group (20 mg/kg), nimodipine group (0.7 mg/kg, positive control group). The occlusion model of middle cerebral artery in rats was established by modified thread occlusion method, and the environment of cerebral ischemia-reperfusion injury was simulated in vivo. Zea longa score was used to detect the neurological deficit of rats after ischemia-reperfusion injury, 2, 3, 5-triphenyltetranitrogen (TTC) was used to detect the volume of cerebral infarction, HE staining was used to detect the pathomorphological changes of nerve cells, Nissl staining was used to observe the changes of nissl bodies, TUNEL staining was used to detect the apoptosis of nerve cells, Western blotting was used to detect the expression of cytochrome C (Cyt C), apoptotic protease activating factor-1 (Apaf-1), Caspase-9 and Caspase-3. Results Compared with the sham group, the neurological deficit symptoms in the model group were significant (P<0.05), the volume of cerebral infarction increased significantly (P<0.05). Under the microscope, it was found that the nerve cells showed contraction of cell body, hyperchromatic and pyknosis of nucleus and poor growth state, the expression of nissl body reduced significantly (P < 0.05), the apoptotic nerve increased significantly (P< 0.05), the expression of Cyt C, Apaf-1, Caspase-9 and Caspase-3 increased significantly (P<0.05). Compared with the model group, the neurological deficit symptoms of calycosin group and nimodipine group reduced significantly (P<0.05), the volume of cerebral infarction reduced significantly (P<0.05). Under the microscope, the damage of nerve cells reduced significantly, the expression of nissl body increased significantly (P<0.05), the apoptotic nerve reduced significantly (P<0.05), the expression of Cyt C, Apaf-1, Caspase-9 and Caspase-3 decreased significantly (P<0.05). Conclusion Calycosin can significantly inhibit the apoptosis of nerve cells and reduce the cerebral ischemia-reperfusion injury. Its mechanism of action is related to the effective regulation of Cyt C/Apaf-1 apoptosis signaling pathway by calycosin.
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Calycosin （CA）， a functional phytoestrogenic isoflavone extracted from Chinese herb Astragali Radix， is characterized by high efficiency， low toxicity， and multiple targets and has multiple pharmacological activities such as anti-oxidation， anti-radiation， anti-bacteria， cardio-cerebrovascular protection， and immunity enhancement. A number of studies have proved its significant anti-tumor effect， making it expected to become a potential component for the treatment of malignant tumors. Research shows that CA exerts the anti-tumor effect via multiple mechanisms like inducing tumor cell apoptosis and inhibiting tumor cell proliferation， migration， and invasion. It has been proved to be effective in suppressing breast cancer， colorectal cancer， lung cancer， cervical cancer， ovarian cancer， nasopharyngeal cancer， and other common malignant tumors. Its anti-tumor activity is mainly related to the regulation of B-cell lymphoma-2 （Bcl-2） family genes， microRNA （miRNA）， and estrogen receptor β （ERβ） to trigger tumor cell apoptosis. Its anti-proliferation activity is mainly reflected in the regulation of cyclin family， WD repeat-containing protein 7 （WDR7-7）， and Ewing sarcoma-associated transcript 1 （EWSAT1）. By blocking the epithelial mesenchymal transformation （EMT）， vascular endothelial growth factor （VEGF）， matrix metalloproteinases （MMPs）， CA inhibits tumor cell metastasis and invasion. In addition， it inhibits tumors by regulating autophagy marker Beclin-1 induced tumor cell autophagy and increases the sensitivity to chemotherapy drugs， thus improving the treatment effect. Although there are many reports about the wide range of applications and good effects of CA in anti-tumor， the systematic review of its anti-tumor mechanism is still lacking. Therefore， this study reviewed the anti-tumor effects and mechanisms of CA， aiming to provide reference for researchers and clinical workers.
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Objective: To establish an HPLC fingerprint spectrum of for decoction pieces-, standard decoction- and dispensing granules of Astragali Radix, and evaluate the correlation among them. Methods: HPLC method was used to determine the content of 7-hydroxy- 4'-methoxy isoflavone, calycosin-7-glucoside, calycosin, and ononin in decoction pieces-, standard decoction- and dispensing granules of Astragali Radix. The common pattern of the three fingerprints was established and the correlation was evaluated through the control fingerprint spectrum. Results: The average contents of 7-hydroxy-4'-methoxy isoflavone, calycosin-7-glucoside, calycosin, and ononin in 20 batches of decoction pieces were 0.097, 0.482, 0.142, and 0.237 mg/g, respectively; The average content of these four index components in the standard decoction were 0.116, 0.912, 0.214, and 0.434 mg/g, respectively; The average content of these four index components in the dispensing granules were 0.088,, 0.623,, 0.160, and 0.289 mg/g, respectively. There were six common peaks in all fingerprint spectra of 20 batches of decoction pieces-, standard decoction- and dispensing granules of Astragali Radix. The similarity between dispensing granules and standard decoction was 0.980, indicating that the components difference between them was very small. Conclusion: The study establishes an HPLC fingerprint spectrum, and itwhich reflects the whole picture of multi-component of decoction pieces-, standard decoction- and dispensing granules of Astragali Radix. It provides reference value for the identification and quality control of Astragalus dispensing granules of Astragalus Radix.
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Objective: To establish a UPLC method to simultaneously determine 10 active ingredients in Yiganning Granules (YG) and provide scientific basis for the quality control, evaluation and standard revision of YG preparations. Methods: A UPLC method was used with an Acquity UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm). The mobile phase was acetonitrile-mehanol-0.15% phosphoric acid solution with gradient elution. The flow rate was 0.3 mL/min. The column temperature was 45 ℃. The injection volume was 2 μL. Results: Ten active ingredients (chlorogenic acid, atractylenolide I, paeoniflorin, calycosin 7-O-β-D- glucopyranoside, stilbene glycoside, caffeic acid, toosendanin, kaempferol, paeonol and tanshinone ⅡA) in YG were simultaneously determined. The linearity was good (r ≥ 0.999 0), the limit of detection and quantification were 0.006-0.017 μg/mL and 0.017-0.510 μg/mL. The average recoveries were 98.8%-102.5% with RSDs of 1.13%-5.37%. Through the determination of 16 batches of samples, the average content of the above 10 ingredients was in turn (5.724 ± 0.017), (0.273 ± 0.003), (0.854 ± 0.005), (1.228 ± 0.004), (0.496 ± 0.003), (1.287 ± 0.004), (0.137 ± 0.004), (3.624 ± 0.014), (7.366 ± 0.032) and (1.754 ± 0.005) mg/g, respectively. Conclusion The established UPLC method is simple, specific, sensitive, stable, precise, accurate, and reproducible, which can be used for quality control and evaluation of YG.
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Objective: To optimize the infiltration process of Astragalus (Astragalus membranaceus var. mongholicus) medicinal materials by Box-Behnken response surface method. Methods: Based on the HPLC-DAD-ELSD and response surface design method, the qualified rate of decoction pieces, the content of index components and bending inspection were used as comprehensive inspection indicators, and the three factors of infiltration were selected for response surface experimental design to optimize the infiltration process of Astragalus medicinal materials parameter. Results: The best infiltration process was as following: infiltration temperature was 20 ℃, with water addition of 1:0.988 for 6 h. Under this process, the qualified rate of Astragalus pieces was 95.81%, the content of calycosin-7-glucoside was 0.072%, and the content of astragaloside IV was 0.276 %. Combining fingerprint analysis and heat map analysis, the material basis of A. membranaceus var. mongholicus changed during the infiltration process. The infiltration parameters should be strictly controlled during the infiltration process to ensure uniform quality of the pieces. Conclusion: The optimized Astragalus medicinal material infiltration process is stable and feasible with good reproducibility, which can provide a reference for the mass production process development of Astragalus medicinal slices.
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Objective: To establish HPLC fingerprints of Qiwei Tangmaishu tablet and simultaneously determine the contents of the six constituents, acteoside, martynoside, calycosin 7-O-β- D-glucopyranoside, schisandrin, formononetin and deoxyschizan- drin. Methods: The analysis of 50% methanol extract of this drug was performed on a 30℃ thermostatic Waters Symmetry C18 column, with the mobile phase comprising of acetonitrile(A)-0.2% formic acid solution(B)flowing at 1.0 ml/min in a gradient elution manner. The detection wavelength was set at 330 nm for acteoside and martynoside, 254 nm for calycosin 7-O-β-D-glucopyranoside, schisandrin, formononetin and deoxyschizandrin. Results: There were thirteen common peaks in the fingerprints of ten batches of samples with the similarities more than 0.9. Acteoside, martynoside, calycosin 7-O-β-D-glucopyranoside, schisandrin, formononetin and deoxyschizandrin showed good linear relationship within the ranges of 1.47-36.75, 0.66-16.50, 0.89-22.25, 2.58-64.50, 1.91-47.75 and 0.77-19.25 μg/ml(r≥0.9993), and their average recoveries were 97.57%, 99.22%, 96.69%, 100.01%, 98.79% and 96.77%, with the RSD of 0.79%, 1.54%, 0.61%, 0.64%, 0.83% and 0.36%, respectively. Conclusion: The established method is easily operable and repeatable, which could be used for quality control of Qiwei Tangmaishu tablet.
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Objective To investigate the effects of calycosin, formononetin, calycosin-7-glucoside and ononin on PC 12 cells differentiation. Methods PC 12 cells were cultured and treated with different concentrations of nerve growth factor (NGF), calycosin, formononetin, calycosin-7-glucoside and ononin for 5 days, once a day, 3 times in a row. The neurite outgrowth of PC 12 cells was observed and the expression of β III-tubulin were measured by immunofluorescence. Results Compared with the vehicle group, neurite outgrowth and the expression of β III tubulin in PC 12 cells had not promoted by calycosin, formononetin, calycosin-7-glucoside and ononin (0.01-10.00 μmol/L). Conclusion PC 12 cells differentiation could not be induced by calycosin, formononetin, calycosin-7-glucoside and ononin.
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Astragali Radix is commonly used as bulk medicinal materials. Chinese Pharmacopoeia contains about 150 compound preparations of Astragali Radix, but the sample preparation method under the determination of Astragali Radix content in Chinese Pharmacopoeia is tedious and time-consuming, not convenient for the test of a large number of samples. Therefore, it is of great significance to simplify the sample preparation method and improve the practicability of the method for the quality control of Astragali Radix and its preparations. In this study, ultrasonic extraction method was used instead of heated reflux extraction, and solid phase extraction method was used to enrich and prepare the samples. A set of practical quality evaluation method was established for Astragali Radix slices and standard decoction, greatly shortening the sample preparation time and improving the accuracy of the method. The results of Astragali Radix standard decoction analysis showed that the transfer rate of calycosin 7-O-β-D-glucospyranoside,(96.5±28.7)%, had great variation, which was found to be related to the conversion of mulberry isoflavone glucoside into calycosin 7-O-β-D-glucospyranoside during the preparation of standard decoction. The transfer rates were(59.4±14.4)% and(101.3±12.3)% for calycosin and astragaloside Ⅳ respectively, which were relatively stable. Therefore, it is suggested that Astragali Radix slices and water decoction preparations should be evaluated by using calycosin and astragaloside Ⅳ as the quality evaluation index. The results provide a scientific and practical method for quality control of Astragali Radix slices and its standard decoction, and also provide scientific evidence for quality evaluation of the preparations.
Subject(s)
Astragalus Plant/chemistry , Astragalus propinquus , Drugs, Chinese Herbal/standards , Glucosides/analysis , Plant Roots/chemistry , Quality Control , Solid Phase ExtractionABSTRACT
Objective To investigate the effect of calycosin on mitochondrial apoptotic pathway in oxygen-glucose deprivation/ reoxygenation PC12 cells. Methods PC12 cells were randomly divided into four groups: control group, model group, calycosin group and nimodipine group. Except for the control group, the other groups were treated with oxygen and glucose deprivation for 2 hours and compound oxygen and glucose for 24 hours. Calycosin group and nimodipine group were treated with drug-containing medium containing calycosin (0. 07 μmol/ L) and nimodipine (5. 00 μmol/ L) simultaneously with reoxygenation. CCK-8 method was used to detect cell survival rate, flow cytometry was used to detect cell apoptosis rate, immunofluorescence method was used to detect Bax/ Bcl-2 ratio, Western blotting was used to detect the expression of key proteins cytochrome C (Cyt-C), apoptotic protease activating factor-1 (Apaf-1) and Caspase-3 in mitochondrial apoptotic pathway. Results Compared with the control group, the survival rate of cells in model group decreased significantly (P<0. 05), and the apoptotic rate increased significantly (P<0. 05), the ratio of Bax/ Bcl-2 was significantly increased (P<0. 05), and the expression of key proteins Cyt-C, Apaf-1 and Caspase-3 in mitochondrial apoptotic pathway were significantly increased (P<0. 05). Compared with the model group, the cell survival rates of calycosin group and nimodipine group increased significantly (P < 0. 05), apoptotic rate decreased significantly (P<0. 05), the ratio of Bax/ Bcl-2 was significantly decreased (P<0. 05), and the expression of key proteins of mitochondrial apoptotic pathway, Cyt-C, Apaf-1 and Caspase-3 were significantly decreased (P < 0. 05). The difference has statistical significance. Conclusion Calycosin can significantly improve the survival rate of oxygen-glucose deprivation/ reoxygenation PC12 cells and inhibit cell apoptosis. Its mechanism is closely related to the inhibition of the expression of key proteins Cyt-C, Apaf-1 and Caspase-3 in mitochondrial apoptotic pathway by calycosin.
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Cardiovascular disease is the main cause of mortality and morbidity in the world, especially in developing countries. Drug therapy is one of the main ways to treat cardiovascular diseases. Among them, great progress has been made in the treatment of cardiovascular diseases with traditional Chinese medicine. In terms of experimental research, the mechanism of traditional Chinese medicine in the treatment of cardiovascular diseases has been thoroughly discussed in vitro and in vivo. In terms of clinical treatment, traditional Chinese medicine with flavonoids, saponins and alkaloids as the main effective components has a definite effect on the treatment of cardiovascular diseases such as arrhythmia, myocardial ischemia, angina pectoris and myocardial infarction, with high safety and good application prospects. With the further research on the effective ingredients, mechanism and adverse reactions of traditional Chinese medicine, it will be beneficial to the effectiveness of traditional Chinese medicine, reduce side effects and promote the modernization of traditional Chinese medicine. Calycosin and its derivatives, the main bioactive flavonoids in Astragalus membranaceus have multiple biological effects, such as antioxidant, pro-angiogenesis, anti-tumour, and anti-inflammatory effects. Based on the above biological effects, calycosin has been shown to have good potential for cardiovascular protection. The potent antioxidant effect of calycosin may play an important role in the cardiovascular protective potential. For injured cardiac myocytes, calycosin and its derivatives can alleviate the cell damage mainly marked by the release of myocardial enzymes and reduce the death level of cardiac myocytes mainly characterized by apoptosis through various mechanisms. For vascular endothelial cells, calycosin also has multiple effects and multiple mechanisms, such as promoting vascular endothelial cell proliferation, exerting vasodilating effect and directly affecting the synthesis function of endothelial cells. The present review will address the bioactivity of calycosin in cardiovascular diseases such as protective effects on cardiac myocytes and vascular endothelial cells and elucidate main mechanism of calycosin and its derivatives to exert the above biological effects.
Subject(s)
Humans , Apoptosis/drug effects , Cardiotonic Agents/pharmacology , Cardiovascular Diseases/drug therapy , Cell Proliferation/drug effects , Drugs, Chinese Herbal/pharmacology , Isoflavones/pharmacology , Medicine, Chinese Traditional , Muscle Cells/drug effectsABSTRACT
The current case–control study sought the association of BDNF rs6265 and MC4R rs17782313 with metabolic syndrome(MetS), MetS components and other related metabolic parameters in a sample of Pakistani subjects. Fasting high-densitylipoprotein cholesterol (HDL-C) and homeostatic model assessment of insulin sensitivity showed a significantly lowermean whereas body mass index (BMI), waist circumference, systolic blood pressure (SBP), diastolic blood pressure (DBP),fasting blood glucose, insulin, total cholesterol (TC), low-density lipoprotein cholesterol, very-low-density lipoproteincholesterol, triglycerides (TG), cholesterol to HDL-C ratio, TG to HDL-C ratio, homeostatic model assessment of insulinresistance, visceral adiposity index, lipid accumulation product and the product of TG and glucose showed a significantlyhigher mean in the presence of MetS. Reduced HDL-C appeared as the most frequent and hypertriglyceridemia as the leastfrequent component of MetS whereas clustering of reduced HDL-C ? abdominal obesity (AO) ? hyperglycemia appearedas the most prevalent combination of MetS components. Moreover, BDNF rs6265 showed BMI and gender independentassociation with increased risk of MetS in Pakistani individuals whereas MC4R rs17782313 showed BMI and genderdependent association with increased risk of MetS in Pakistani females. In addition, BDNF rs6265 and MC4R rs17782313showed gender-dependent associations with decreased risk of having low HDL-C in males and increased risk of havingabdominal obesity in females, respectively. However, no association was observed for metabolic variables other thancomponents of MetS across genotypes of both BDNF rs6265 and MC4R rs17782313.