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Objective:To investigate the protective effect and mechanism of isorhamnetin on myocardial injury in rats with acute myocardial infarction (AMI).Methods:The AMI rat model was established by coronary artery left anterior descending ligation. The SD rats were divided into sham-operated group, model group, Fasudil group (30 mg/kg) and low-, medium-, and high-dose (25, 50, 100 mg/kg) groups. Each group had 5 rats. They were administrated intragastric, once a day, and were treated for 14 d. Color Doppler ultrasonography was used to detect cardiac function by measuring left ventricular end-systolic diameter (LVESd), left ventricular end-diastolic diameter (LVEDd), and left ventricular long-axis shortening fraction (FS). Myocardial infarct size was detected by TCT staining. Superoxide dismutase (SOD), malondialdehyde (MDA), interleukin (IL)-6, IL-1β, and tumor necrosis factor-α (TNF-α) levels in rat serum were detected by ELISA and TUNEL assay for the cardiomyocyte apoptosis index. Cysteinyl aspartate specific proteinase-3 (Caspase-3) activity in myocardial tissue was detected by the Caspase-3 activity assay kit, and the expression of Bcl-2 associated X protein (Bax) and B-cell lymphoma-2 (Bcl-2) proteins in myocardial tissue was detected by Western Blot.Results:Compared with the sham operation group, the FS, SOD content, and expression levels of Bcl-2 protein in the model group were significantly decreased (all P<0.05), and LVEDd, LVESd, myocardial infarct size, MDA content, IL-6 content, TNF-α content, IL-1β content, apoptosis index, Caspase-3 activity, and Bax protein expression level were significantly increased (all P<0.05). There was no significant difference between the low concentration of isorhamnetin and the model group (all P>0.05). However, the above changes caused by the construction model after treatment with Fasudil and medium- and high-concentrations of isorhamnetin significantly reversed (all P<0.05). Conclusions:Isorhamnetin can improve cardiac function and protect myocardial injury in AMI rats by reducing oxidative stress and inflammatory damage to cardiomyocytes and inhibiting cardiomyocyte apoptosis.
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Objective: To explore the active compounds of Huoxiang Zhengqi Oral Liquid for the treatment of coronavirus desease 2019 (COVID-19) by network pharmacology and molecular docking. Methods: The chemical constituents and action targets of Atractylodes chinensis, Citrus reticulata, Magnolia officinalis, Angelica dahurica, Poria cocos, Areca catechu, Pinellia ternata, Glycyrrhiza uralensis, Pogostemon cablin and Perilla frutescens were retrieved from TCMSP. Uniprot database was used to search the corresponding genes of targets, then Cyoscape 3.7.2 software was used to construct the network of medicinal materials-compound-target (gene) for visualization; GO function enrichment analysis and KEGG pathway enrichment analysis were performed through DAVID to predict its mechanism of action, and histograms and bubble maps were plotted by Prism software and Omicshare database for visualization. Results: The network of medicinal materials-compound-target contained 10 medicinal materials, 123 compounds and 257 corresponding target genes, and the key target genes involved PTGS2, HSP90AB1, AR, CAMSAP2, PPARG, NOS2, etc. GO functional enrichment analysis resulted in 278 GO entries (P < 0.05), including 178 biological processes (BP) entries and 36 cellular component (CC) entries, and 64 molecular function (MF) entries. KEGG pathway enrichment analysis revealed that there were 119 (P < 0.05) signaling pathways involving Hepatitis B, small cell lung cancer, non-small cell lung cancer, bladder cancer, prostate cancer and T cell receptor pathways. The results of molecular docking showed that the core compounds such as quercetin, isorhamnetin, irisolidone, kaempferol, wogonin, and baicalein were similar in affinity with the COVID-19 recommended medicine. Among them, quercetin, isorhamnetin and irisolidone had the strongest affinity. Conclusion: The compounds in Huoxiang Zhengqi Oral Liquid can combine with angiotensin converting enzyme II (ACE2) binding to PTGS2, HSP90AB1, AR, CAMSAP2 and other targets to regulate multiple signaling pathways, thus exerting a preventive or therapeutic effect on COVID-19.
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Objective: To find out the active compounds of Yupingfeng San for the prevention and treatment of the coronavirus pneumoniadisease 2019 (COVID-19) using network pharmacology and molecular docking, with a purpose to find a better clinical use of Yupingfeng San. Methods: The effective ingredients and targets of Astragali Radix, Atractylodis Macrocephalae Rhizoma and Saposhnikoviae Radix were searched from the traditional Chinese medicine system pharmacology analysis platform (TCMSP) website, and the protein and protein interactive network of Yupingfeng San was established using the String database (PPI). Cytoscape 3.6.1 software was used for data analysis to extract the Hub network from the PPI network. The KEGG pathway analysis was performed using the String database and the molecular docking was performed using ChemOffice, PyMOL, and Auto Dock software. Results: A total of 45 effective ingredients were obtained with limited screening conditions [oral bioavailability (OB) ≥ 30%; drug-like (DL) ≥ 0.18], and 345 potential targets and 15 key targets of Yupingfeng San were screened. A total of 50 pathways were obtained by KEGG pathway analysis, among which 25 main pathways were selected, including PIK3R1 target regulation PI3K-Akt signaling pathway, Ras signaling pathway, HIF-1 signaling pathway, and MAPK signaling pathways and T cell receptor signaling pathways. Conclusion: The active compounds in Yupingfeng San can inhibit the combination between SARS-CoV-2 protein and angiotensin-converting enzyme II (ACE2), thus regulating multiple signal pathways (PIK3R1, IGF1R, etc), which plays a role in the prevention of COVID-19.
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Objective: To study the chemical constituents of Empetrum nigrum var. japonicum. Methods: The chemical constituents were isolated by silica gel column chromatography and HPLC, and its structure was identified by their spectral data and physicochemical properties analysis. Results: Twenty-two compounds were isolated from ethyl acetate extracts of E. nigrum var. japonicum with the structures identified as 3,4-dihydroxybenzoic acid (1), 3-methoxy-4-hydroxybenzoic acid (2), methyl-3,4-dihydroxybenzoate (3), dihydroconiferyl alcohol (4), (6S,9R)-vomifoliol (5), (6R,9R)-9-hydroxy-3-one-α-ionol (6), phenylpropionic acid (7) quercetin-3-O-(6″-benzoyl)-β-D-galactoside (8), daucosterol (9), quercetin-3-O-β-D-glucopyranoside (10), phenylpropyl-O-β-D-glucopyranoside (11), maslinic acid (12), rutinum (13), quercetin-3-O-β-D-galactoside (14), benzyl-(6-O- α-L-arabinofuranosyl)-O-β-D-glucopyranoside (15), cinnamol-O-(6-O-α-L-arabinopyranosyl)-β-D-glucopyranoside (16), quercetin-3- O-neohesperidoside (17), kaempferol-3-O-neohesperidoside (18), 2α,3β-dihydroxyurs-12-en- 28-oic acid (19), foliachinenoside A2 (20), kaempferol-3-O-rutinoside (21) and isorhamnetin-3-O-rutinoside (22). Conclusion: Compounds 1-6, 8, 11-13, 15-22 were isolated from E. nigrum var. japonicum for the first time.
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Objective: Based on the Chinese herbal property, the pharmacodynamic evaluation and mechanism discussion of active components were carried out through screening the active components of Chinese patent medicine in the clinical medical insurance catalog. Methods: The most frequently used main herbs were screened through the collection of anti-vitiligo Chinese patent medicine prescriptions, drug properties and material basis. The main compound types were acquired through TCMSP and TCMIP databases. The drug properties were analyzed by admetSAR method to obtain key compounds. The pharmacodynamics were observed by measuring the morphology and melanin content of adult zebrafish and larvae. The safety evaluation was indicated by the survival rate of larvae. RT-PCR was used to reveal the mechanism of the compounds at the transcriptional level. The binding ability of compounds to protein crystal structure was predicted by molecular docking. Results: The most frequently used main herbs were Carthamus tinctorius, Lithospermum erythrorhizon, Tribulus terrestris, Gentianae Radix et Rhizoma., Psoraleae Fructus, and Vernonia anthelmintica. The main compound types through TCMSP and TCMIP database were flavonoids with a total of 81. Based on the druggability and stability, the methoxyflavones kaempferide and isorhamnetin were screened out. Kaempferide (32 μmol/L), isorhamnetin (32 μmol/L) and methoxsalen (25 μmol/L) could promote the regeneration of melanin in zebrafish. Based on the zebrafish embryo model, kaempferide, isorhamnetin and methoxsalen all could accelerate melanogenesis in larvas, and the survival rates of larvas were more than 90% under effective concentration. RT-PCR showed that kaempferide and isorhamnetin upregulated the mRNA levels of MC1R and MITF genes related to melanogenesis. The results of molecular docking between the structures of proteins (MITF, TYR, TYRP1) and kaempferide, isorhamnetin, methoxsalen showed that the binding score of kaempferide or isorhamnetin was higher than that of methoxsalen. Conclusion: Kaempferide and isorhamnetin, the active ingredients in the clinical anti-vitiligo traditional Chinese medicine prescriptions, can promote the melanogenesis in zebrafish by up-regulating the MC1R/MITF signal pathway.
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OBJECTIVE：To compare the quality between Stamen typhae and pollen of T. angustifolia ，and provide scientific evidence for the improvement of quality standard of T. angustifolia . METHODS ：Fifteen batches of S. typhae were collected. Pollen minus sieve ，impurity plus sieve （filament and anther ）were sift out from S. typhae according to the identification method of T. angustifolia in Chinese Pharmacopoeia （2015 edition）. The characteristics and components of S. typhae and pollen ，filament and anther of T. angustifolia were comfirmed by impurity ， character examination and microscope ， TLC. The contents of isorhamnetin-3-O-neohesperidoside and typhaneoside in S. typhae and pollen ，impurities plus sieve （filament and anther ）of T. angustifolia were determined by HPLC. RESULTS ：S. typhae was a mixture of pollen ，anther and filament of T. angustifolia ，in the form of brownish yellow flocculent. The pollen of S. typhae was yellow powder with delicate hand feel ，slight smell and light taste；the surface of cells was slightly striped. The filaments and anthers were filiform and short-term ，rough and astringent ，and the cell surface were long strip. TLC chromatogram of S. typhae ，pollen and impurity of T. angustifolia had the same color spots at the same location. The contents of isorhamnetin- 3-O-neohesperidoside，typhaneoside and their aggregate were the highest in pollen （0.42％，0.24％，0.64％）；the second in S. typhae （0.22％，0.17％，0.39％）；the lowest in the impurities plus sieve （0.19％， 0.14％，0.33％）. The total contents of isorhamnetin- 3-O-neohesperidoside and typhaneoside in S. typhae and in impurities plus sieve did not reach the content limit stipulated in Chinese Pharmacopoeia （not less than 0.50％）. CONCLUSIONS：The medicinal components of T. angustifolia mainly exist in pollen. It is suggested that S. typhae should be used as the raw material to obtain pollen，and should not be used directly.
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Aim To investigate whether isorhamnetin could protect against rotenone-induced PC 12 cells injury via regulating PI3K/Akt/GSK-3p/CREB pathway. Methods PC 12 cell viability was determined by MTT and LDH assays. The expressions of Akt,p-Akt,GSK-3p,p-GSK-3p,CREB and p-CREB were measured by Western blot. Results The cell viability and level of phospho-CREB significantly decreased in PC 12 cells exposed to rotenone when compared to control group. Both the cell viability and the expression of phospho-CREB in cells pretreated with isorhamnetin were higher than those of cells exposed to rotenone alone. Moreover, pretreatment of PC 12 cells with isorhamnetin enhanced phosphorylation of Akt and GSK-3fi. The addition of LY294002 could suppress the phosphorylation of Akt, GSK-3p and CREB, resulting in abolishment of neuroprotective effects of isorhamnetin on cells exposed to rotenone. Conclusion Isorhamnetin might alleviate rotenone-induced PC 12 cells injury partially via the PI3K/Akt/GSK-3 (3/CREB signaling pathway.
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We studied the protective effect and mechanism of isorhamnetin (ISO) on 1-methyl-4-phenylpyridiniumion (MPP+)-induced SH-SY5Y cells injury. MPP+-induced SH-SY5Y cell injury model was established, and cell viability was measured by MTT and LDH methods. The activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in cells were determined to investigate the level of oxidative stress. DCFH-DA and MitoSOX fluorescence probes were used to detect the levels of intracellular reactive oxygen species (ROS) and mitochondria superoxide, respectively. JC-1 fluorescence probe was used to detect the changes of mitochondrial membrane potential. Western blot and immunofluorescence methods were used to determine the expressions of Sirt1 and PGC-1 proteins, as well as the expression levels of apoptosis-related proteins Bax and Bcl-2. MPP+ at the dose of 500 μmol·L-1 significantly reduced SH-SY5Y cells viability to 52.46% and increased LDH release to 417.63%. ISO at 5 and 15 μmol·L-1 significantly increased the expression of Sirt1 and PGC-1α, inhibited LDH release, reduced intracellular ROS and mitochondria superoxide, inhibited the decline of mitochondrial membrane potential and increased cell viability to 61.61% and 67.55%. In addition, ISO could downregulate the expression of Bax and upregulate the expression of Bcl-2 to reduce cell apoptosis. ISO-mediated inhibition of apoptosis could be reversed by Sirt1 specific inhibitor Sirtinol. Through activating Sirt1/PGC-1α signaling pathway, ISO could reduce oxidative stress injury and inhibit cell apoptosis to protect cells from MPP+ injury.
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This study aimed to explore the anti-tumor activity and mechanisms of action of isorhamnetin, a compound isolated from Astragalus membranaceus, in combination with sorafenib for treatment of renal cell carcinoma (RCC). The anti-tumor activity of isorhamnetin in combination with sorafenib was detected by MTT assay with cells in culture or Renca xenograft model in mice. Western blot was used to study the mechanisms of isorhamnetin in combination with sorafenib. Lymphocyte proliferation assay was also used to investigate the effects of the two drugs in combination. The results indicated that isorhamnetin inhibited the proliferation of RCC cells, with IC50 for A498, 786-O and Renca cell lines with being 31.7, 28.8 and 106.0 μmol·L-1, respectively. Isorhamnetin in combination with sorafenib improved the anti-lymphocyte proliferation activity of sorafenib with the IC50 down to 12.0 μmol·L-1. Isorhamnetin inhibited the growth of RCC in mice slightly with the inhibition efficiency at 26.9%. With 50.0 mg·kg-1 isorhamnetin in combination with 20.0 mg·kg-1 sorafenib, the anti-tumor activity of sorafenib was enhanced, with inhibition of growth rate increased to 60.7%. Meanwhile, isorhamnetin in combination with sorafenib could promote the lymphocytes proliferation in Renca xenograft model. Western blot results showed that combination of isorhamnetin and sorafenib could inhibit c-Raf/MEK/ERK and AKT/mTOR signaling pathways. In conclusion, the combination of isorhamnetin with sorafenib could increase the anti-tumor activity of sorafenib in RCC in vitro and in vivo. The mechanisms may be related to the inhibition of c-Raf/MEK/ERK and AKT/mTOR signaling pathways. Procedures for animal study were performed with approval of the Animal Care and Use Committee of the Chinese Academy of Medical Sciences and Peking Union Medical College.
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Objective: To establish a UPLC-MS/MS analysis method for determination of baicalin, geniposide, chlorogenic acid, cholic acid and hyodeoxycholic acid in Qingkailing (lyophilized) for injection in rat plasma, and to investigate the pharmacokinetic behavior of this preparation in normal and cerebral ischemic rats. Method: Rats were randomly divided into normal group and cerebral ischemia model group. The rat model of cerebral ischemia was established by suture embolization. The rats were given by intraperitoneal injection, and normal saline was used as the solvent. Blood samples were taken at the corresponding time points. After treatment, UPLC-MS/MS was used to determine the blood concentration of five components. The main detection conditions were mobile phase of 0.1%formic acid aqueous solution-acetonitrile for gradient elution (0-0.25 min, 90%A; 0.25-1 min, 90%-75%A; 1-2 min, 75%-50%A; 2-2.6 min, 50%-45%A; 2.6-2.65 min, 45%-90%A; 2.65-4.0 min, 90%A), the flow rate of 0.4 mL·min-1, the column temperature at 40℃, electrospray ionization under negative ion mode. The pharmacokinetic parameters were fitted and the bioavailability was calculated, the differences of treatment process of five components from Qingkailing (lyophilized) for injection in normal and cerebral ischemic rats were analyzed. Result: Compared with the normal group, the area under the curve (AUC0-t) of geniposide in rats from cerebral ischemia model group decreased significantly after intraperitoneal injection of Qingkailing (lyophilized) for injection (PTmax) of chlorogenic acid in rats from cerebral ischemia model group was significantly earlier than that in the normal group (PConclusion: Qingkailing (lyophilized) for injection has a certain difference in the treatment process between normal and cerebral ischemic rats, which has certain guiding significance for the clinical treatment of cerebral ischemic diseases with this preparation.
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Objective To investigate the relationship between the content of flavonoids and the key enzyme genes expression in different tissues of Bupleurum chinense and B. scorzonerifolium. Methods The roots, stems, leaves, and fruits of B. chinense and B. scorzonerifolium were used as test materials, determination of flavonoids (rutin, quercetin, kaempferol, and isorhamnetin) in different tissues by HPLC, determination of total flavonoids by UV spectrophotometry, the tissues expression of key enzyme genes (IFS, F3H, and DFR) in flavonoids synthesis was determined by real-time quantitative PCR, correlation analysis was performed with SPSS. Results The content of flavonoids in the aerial parts (stems, leaves, and fruits) of B. chinense and B. scorzonerifolium was significantly higher than that in roots, the content of flavonoids was mainly rutin, and the content of rutin in the leaves of B. chinense leaves was up to 106.961 mg/g; The distribution of total flavonoids in B. chinense and B. scorzonerifolium was obviously different, the content was from high to low: leaves ≥ fruit > stem > root; The expression of B. chinense IFS, F3H, and DFR gene in the aerial parts was much higher than that in roots, IFS gene was significantly positive correlated with rutin (P < 0.05), F3H gene was significantly positive correlated with DFR gene (P < 0.05), but the expression of IFS, F3H, and DFR gene in each tissues of B. scorzonerifolium was at lower level. Conclusion The content of flavonoids in different parts of B. chinense and B. scorzonerifolium was consistent with the expression of flavonoids synthesis key enzyme genes, the differential expression of key enzyme genes regulates the synthesis and accumulation of flavonoids in different tissues.
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OBJECTIVE To study transmembrane transport and methylation metabolism of quercetin based on the human intestinal absorption model. METHODS Caco-2 cell monolayer was used as the human intestinal absorption model. The incubation concentration of quercetin was 9.0 and 18.0 mg-L'1. Samples were collected at 30, 60, 90, 120 and 150 min, and the contents of quercetin, isorhamnetin and tamarixetin on the loading side and receiver side were determined by high performance liquid chromatography-mass spectrometry (LC-MS) method. Bupropion was used as the internal standard, and the injection volume was 20 pL. The specific inhibitors of P-glycoprotein (P-pg) or multidrug resistant protein 2 (MRP2), cyclosporins (CysA) 10 mmol-L-1 or MK571 1 mmol • L-1 (final concentration), were added to the apical side. After 15 min incubation, quercetin 9.0 or 18.0 mg-L-1 (final concentration) was added to the apical side, respectively. The quercetin content on the receiver side was determined by the same method. RESULTS During bi-directional transport, the dynamic change of quercetin residues on the loading side showed a continuous decline within 150 min (P<0.05 between adjacent time points), while the amount of quercetin on the receiver side tended to increase before decreasing, reaching the peak at 120 min and falling at 150 min (P<0.05 between adjacent time points). Isorhamnetin and tamarixetin could be detected on both the loading side and receiver side. But the difference was that when quercetin was loaded on the apical side or the basolateral side, there was much more isorhamnetin and tamarixetin on the apical side than on the basolateral side after 30-60 min. Analysis of the percentage of quercetin, isorhamnetin and tamarixetin on the loading side and receiver side found that when incubated for 30 min, the residual quercetin on the loading side was less than 20%-25%, and quercetin on the receiver side was only about 1% of the loading amount. At 150 min, the residual quercetin decreased to <10%, while quercetin in the receiver side was only 6%-7% of the loading amount, and isorhamnetin and tamarixetinin on both sides were only 0.1%-0.3% of loading quercetin. Compared with the quercetin control group, the addition of CysA or MK571 in advance significantly increased the transport of quercetin from the apical side to the basolateral side (P<0.05). CONCLUSION The transport of quercetin on the Caco-2 monolayer cell model shows a trend of rise and fall, accompanied by methylation metabolism. The efflux of P-pg and MRP2 may have an effect on the transmembrane transport of quercetin.
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Objective @#To investigate the effect and potential molecular mechanisms of isorhamnetin (ISO) extracted from Ginkgo biloba on the differentiation of osteoclasts.@*Methods@#Osteoclast precursor RAW264.7 cells were induced with RANKL to differentiate into mature osteoclasts. Different concentrations of ISO were added to RAW264.7 cells to determine its effect on osteoclast differentiation. CCK8 was used to evaluate the effect of ISO on cytotoxicity. The impact of ISO on the osteoclast differentiation process was investigated by analyzing tartrate resistance and bone resorption lacuna. Real-time PCR was performed to analyze the levels of differentiation marker genes, including tartrate resistant acid phosphatase (Trap), cathepsin K (Ctsk), and matrix metalloproteinase 9 (MMP-9); differentiation-related transcription factors, including the proto-oncogene protein c-Fos, nuclear factor of activated T-cells cytoplasmic 1(NFATc1); and the levels of downstream NF-κB p65 signaling pathway phosphorylation. Using the above-described method, we verified that ISO exerted an inhibitory effect on osteoclast differentiation and explored related molecular mechanisms. @*Results @#Different concentrations of ISO (1-10 μM) had no cytotoxic effects on RAW264.7 cells, inhibited TRAP activity and decreased the number of bone resorption lacuna during osteoclast differentiation. When applied at a concentration of 10 μM, its inhibitory effect was significant. In addition, ISO significantly reduced the expression levels of Trap, Ctsk, MMP-9, c-Fos, NFATc1 and NF-κB p65 mRNA. @*Conclusion@# ISO extracted from Ginkgo biloba extract exerted an inhibitory effect on osteoclast differentiation, and the mechanism underlying its activity may involve the inhibition of the classical NF-κB pathway.
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The present study was conducted to explore the effect of light intensity on growth, bioactivity compounds accumulation and anti-oxidative activity of Sedum sarmentosum. The growth, yield, contents of total flavonoids, total phenolic, quercetin, kaempferol and isorhamnetin, and antioxidant activities were assessed in S. sarmentosum under five light intensities, namely 100% full sunlight (G1), 77% full sunlight (G2), 60% full sunlight (G3), 38% full sunlight (G4), and 16% full sunlight (G5). The results showed that light intensity significantly affected the growth and the chemical compounds accumulation. With the decrease of light intensity, the maximum branch length and the average internode distance increased. G2 treatment greatly promoted the numbers of leaf layers and branches, and G3 treatment remarkably improved the yield. The highest total flavonoids and phenolic contents were obtained in G3 treatment. Meanwhile, the highest quercetin and isorhamnetin contents were obtained in G1 treatment. The difference of kaempferol content was not significant. In addition, based on DPPH, FTC and FRAP methods, the antioxidant activities of the aqueous extracts under G1 treatment were superior to the others. The results indicated that more than 60% full sunlight was the optimum light intensity condition to achieve high yield and quality of S. sarmentosum.
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Antioxidants , Flavonoids , Phenols , Plant Extracts , SedumABSTRACT
Objective: To establish an HPLC method for the simultaneous determination of 3 active components in compound he-mostatic capsules and investigate the content changes before and after 60Co-γ ray irradiation. Methods: An Agilent-C18column (250 mm×4. 6 mm, 5 μm) was adopted and the flow rate was 1. 0 ml·min-1. The mobile phase consisted of acetonitrile-0. 5% phosphor-ic acid solution (triethylamine was used to adjust pH to 6) with gradient elution, the detection wavelength was 254 nm (0-35 min) and 281 nm (35-55 min), and the column temperature was 35℃. The compound hemostatic capsules were irradiated at 5, 8 and 10 kGy, respectively, and the contents of the 3 active components in compound hemostatic capsules were compared before and after the radia-tion,and the t-test was applied to investigate the significance. Results: Typhaneoside,isorhamnetin-3-O-neohesperidin and tetrahydro-palmatine respectively within the concentration range of 0. 018-0. 11 mg·ml-1,0. 020-0. 120 mg·ml-1and 0. 011-0. 066 mg·ml-1 showed a good linear relationship with the peak area. The average recovery was 97. 7% , 98. 7% and 99. 3% with the RSD of 0. 9% , 0. 9% and 0. 5% , respectively (n=6). After the irradiation, the contents of the three active components in compound hemostatic capsules all showed changes. Under the dose of 8 kGy, the content changes of typhaneoside and isorhamnetin-3-O- neohesperidin showed no statistical significance (P>0. 05),and when the dose increased to 10 kGy,the content of tetrahydropalmatine exhibited sig-nificant difference (P<0. 05). Conclusion: The established determination method for compound hemostatic capsule shows such ad-vantages as high recovery, good repeatability, simple operation and promising separation, which can be used as a quality control meth-od for compound hemostatic capsules. The sterilization effect of 60Co-γ ray irradiation on compound hemostatic capsules is significant without notable changes in the active components. When the irradiation dose is equal to or below 5 kGy,the change of each component is not obvious,which can provide reference for the radiation sterilization of compound hemostatic capsules.
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Objective To study the quality standard of the imitation ecological planting Anoectochilus roxburghii under forest covering. Methods The identification of medicinal properties, microscopic characteristics, thin-layer chromatography (TLC) and content determination of A. roxburghii cultivated under forest were carried out in this study, and the moisture content, ash and acid-insoluble ash were determined according to Chinese Pharmacopoeia of 2015 edition. Results A. roxburghii cultivated under forest exhibits specific properties in characteristics, microscopic features and TLC results. The average moisture content of A. roxburghii cultivated under forest from five planting bases was 8.69%, the average ash was 11.93%, and the average acid-insoluble ash was 3.27%. Content determination results of the average quality score of quercetin, isorhamnetin and kaempferide were at 0.021 0%,,0.024 7%, and 0.027 3%, respectively. Conclusion The above method is simple, specific and reproducible, which will provide the basis for the quality standard of the imitation ecological planting A. roxburghii under forest covering.
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Objective: To establish the HPLC specific chromatograms of the ethyl acetate layer in ten batches of effective parts of Filifolium sibiricum and to determine the contents of five components. Methods: The analysis of effective parts of F. sibiricum was performed on a Thermo AcclaimTM120 C18 column (250 mm × 4.6 mm, 5 μm) with acetonitrile (B)-PBS (A) (0.1 mol/L sodium dihydrogen phosphate-2% glacial acetic acid, 1∶1) as the mobile phase in a gradient elution mode, the detection wavelength was set at 360 nm, the flow rate was 0.8 mL/min, and the column temperature was 35 ℃. Results: The specific chromatograms of F. sibiricum effective parts were established and ten common peaks were designated. Among them, five components including isorientin, isovitexin, isoquercitrin, luteoloside and isorhamnetin-3-O-β-D-glucose all showed good linear relationship within the ranges of 0.018—0.108, 0.066 8—0.400 8, 0.088—0.528, 0.118 4—0.710 4 and 0.017 6—0.105 6 μg, respectively. The average recovery was 98.67%, 97.93%, 98.95%, 99.81%, and 97.33% with the RSD value at 1.10%, 0.93%, 1.10%, 0.62%, and 1.48%, respectively. Moreover, the similarity of the eight batches of samples was above 0.9 in the ten batches of medicinal herbs, the similarity of the two batches of which was 0.688 and 0.695, indicating that its content was lower and the difference was greater. In addition, there were significant differences in the content of five components in each harvest time. The content of flavonoids in medicinal herbs was higher with high flower percentage. It was suggested that the content of flavonoids in F. sibiricum was related to the flower percentage of harvest period. Conclusion: The HPLC specific chromatograms of the F. sibiricum effective parts were established and the common characteristic peaks were determined, which could be used for quality control of the F. sibiricum.
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Objective To establish the identification method of Typha angustifolia (stir-bake to black), Sophora japonica, Sanguisorba officinalis (stir-bake to black), and Rheum palmatum in Fuhuang tablets. Methods HPLC method were studied to identify typhaneoside and isorhamnetin-3-O-neoheptanoside in T. angustifolia, and sophoricoside in S. japonica. HPLC conditions were as follows: Agilent TC-C18 (2) column (250 mm × 4.6 mm, 5 μm) with a gradient elution at a temperature of 30 ℃; Phosphate buffer and acetonitrile were used as the mobile phase with a flow rate of 1.0 mL/min; The detection wavelength was 254 nm. The acid hydrolysis-HPLC method was explored to identify S. officinalis (stir-bake to black). TLC identification method was performed to identify R. palmatum. Results HPLC method can identify typhaneoside, isorhamnetin-3-O-neoheptanoside, and sophoricoside synchronously. Acid hydrolysis method can identify S. officinalis (stir-bake to black) by HPLC and TLC can identify R. palmatum. Conclusion The three methods are simple and accurate with high sensitivity and good specificity, which can be used to identify all herbal medicines in Fuhuang tablets.
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Objective: To explore the effect of soil moisture on growth, bioactivity compounds accumulation, and anti-oxidative activity of Sedum sarmentosum. Methods: The changes of growth, yield, contents of total flavonoids, total phenolic, quercetin, kaempferol and isorhamnetin, and anti-oxidant activities were assessed in S. sarmentosum under five water gradient, namely 15% - 20% FC (field capacity, S1), 35%-40% FC (S2), 55%-60% FC (S3), 75%-80% FC (S4), and 95%-100% FC (S5). Results: S4 treatment greatly promoted the growth and yield while severe drought suppressed growth. The highest total flavonoids content was obtained in S4 treatment, while the lowest was found in S5 treatment. The total phenolic content in S3 treatment was the largest, but there was no significant difference comparing with S4 treatment. Meanwhile, the highest quercetin, kaempferol, and isorhamnetin content were obtained from S1, while S5 treatment showed the lowest values. Besides, the yield of three flavone ingredients per plant peaked in S4 treatment, followed by S5 treatment, and the S1 treatment resulted in the smallest yields. In addition, S4 treatment resulted in the highest anti-oxidant activities of the aqueous extracts based on DPPH, TBARS, and FRAP methods. There was a significantly positive relationship among the antioxidant activities of the aqueous extracts based on TBARS, contents of quercetin, isorhamnetin. Conclusion: In summary, the Results: indicated that 75%-80% FC was the optimum soil moisture condition to achieve high yield and quality of S. sarmentosum.
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Objective To study the chemical constituents of Gendarussa vulgaris. Methods The chemical constituents were isolated and purified with silica column chromatography and gel chromatography, etc. Their structures were identified by physico- chemical properties and various spectroscopic methods including NMR spectrum, MS, UV, etc. Results A total of 16 compounds were isolated and elucidated as daucosterol (1), 6″-O-acetylisavitexin (2), 9,10-dihydroxy-4,7-megastigmadien-3-one (3), 22E,24R-ergosta- 7,22-diene-3β,5α,6β,9α-tetraol (4), isorhamnetin (5), quercetin (6), eleutheroside E (7), gusanlung A (8), gusanlung B (9), betulin (10), isovitexin-2″-O-rhamnoside (11), isovitexin (12), genkwanin (13), apigenin (14), quercetin 3-O-β-D-glucuronide (15), and 2-(4-hydroxy-3-methoxyphenyl)-3-(2-hydroxy-5-methoxyphe-nyl)-3-oxo-1-propanol (16). Conclusion Compounds 3, 5, 11, 12 and 15 are isolated from G. vulgaris for the first time. Compound 2, 4, 13, and 16 are isolated from the plant of genus for the first time.