Protective effects of diosmetin on renal ischemia-reperfusion injury in mice / 现代泌尿外科杂志

【Objective】 To investigate whether diosmetin has protective effects on renal ischemia-reperfusion (I/R) injury in mice and explore the potential mechanism. 【Methods】 A total of 45 BALB/c male mice were randomly divided into the sham group, I/R group and diosmetin treatment group. The protective effects of diosmetin on the renal function of mice was evaluated by detecting blood biochemical indexes and renal tissue HE staining injury score. The expression of p-P65, a key molecule of NF-κB family, was detected with immunohistochemistry and western blotting, and the downstream inflammatory factors of NF-κB signaling pathway were detected with quantitative polymerace chain reaction (qPCR). 【Results】 Compared with the sham group, the I/R group had significantly increased levels of serum creatinine and urea nitrogen, significantly damaged morphological construction, and significantly increased expressions of p-P65 and inflammatory cytokines, while the diosmetin treatment group had decreased levels of serological indexes, improved structure of kidney tissue and reduced expressions of p-P65 and inflammatory cytokines. 【Conclusion】 Diosmetin can protect kidney from I/R-induced acute kidney injury by inhibiting NF-κB pathway in mice.

Effect of diosmetin on ferroptosis of hippocampal cells in rats with bacterial meningitis and the mechanism of SIRT1-Nrf2 signaling pathway / 中华行为医学与脑科学杂志

Objective:To explore the effect and mechanism of diosmetin (Dio) on neuronal ferroptosis in rats with bacterial meningitis (BM).Methods:Male SD rats aged 6-7 weeks of SPF grade were selected for the experiment. The BM model was established by injecting group B hemolytic streptococcus into the cisterna magna of cerebellum. Sixty BM model rats were successfully modeled and divided into model group, low-dose Dio group, medium-dose Dio group, high-dose Dio group and inhibitor group according to the random number table method, with 12 rats in each group. Another 12 weight-matched rats were taken as the control group.The rats in the low-dose Dio group, medium-dose Dio group, high-dose Dio group and the inhibitor group were intragastrically administered with Dio at 50 mg/kg, 100 mg/kg, 200 mg/kg and 200 mg/kg, respectively. The rats in the control group were intragastrically administered with an equal volume of 0.9 % sodium chloride solution. On the day of intragastric administration, the rats in the inhibitor group were intraperitoneally injected with SIRT1 pathway inhibitor EX527 (10 mg/kg), and the rats in the other groups were injected with an equal volume of 0.9% sodium chloride solution. The above interventions were performed once a day for 28 consecutive days. Loeffler neurological score was used to evaluate the neurological impairment in rats. Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in cerebrospinal fluid of rats were detected by ELISA. The number of white blood cells in cerebrospinal fluid was detected by a blood cell analyzer. Glutathione (GSH) was detected by micro-enzyme labeling method, malondialdehyde (MDA) was detected by thiobarbituric acid colorimetric method, reactive oxygen species(ROS) was detected by colorimetry, and Fe 2+ level was detected by ferrozine method. Hematoxylin-eosin staining, Prussian blue staining and TUNEL staining were used to observe the pathological damage, iron accumulation and apoptosis in the hippocampus, respectively.Western blot was applied to measure the expression of transferrin (Tf), proliferating cell nuclear antigen (PCNA), Bcl-2-associated X protein (Bax), caspase-3 and SIRT1/Nrf2/HO-1/Gpx4 signaling pathway proteins. Graphpad Prism 9.0 was used for data analysis. One-way ANOVA was used for statistical analysis, and SNK- q test was used for further pairwise comparisons. Results:(1) There was a statistically significant difference in neurological function scores among the 6 groups of rats ( F=125.451, P<0.001). The neurological function score of the model group was lower than that of control group, while the neurological function scores of the low-dose Dio group, medium-dose Dio group, and high-dose Dio group were higher than those of the model group (all P<0.05). The neurological function score of the inhibitor group ((2.57±0.26)) was lower than that of high-dose Dio group ((4.34±0.48)) ( P<0.05). (2) There were statistically significant differences in the levels of IL-6, TNF-α and the number of white blood cells in the cerebrospinal fluid of rats among the 6 groups ( F=127.817, 102.413, 180.967, all P<0.001). The levels of IL-6, TNF-α and the number of white blood cells in model group were higher than those of control group(all P<0.05). The levels of IL-6, TNF-α and the number of white blood cells in low-dose Dio group, medium-dose Dio group and high-dose Dio group were lower than those of model group (all P<0.001), and those in inhibitor group were all higher than those in high-dose Dio group(all P<0.001). (3) There were statistically significant differences in iron deposition rate and neuronal apoptosis rate among the 6 groups of rats ( F=90.857, 88.835, both P<0.001). The iron deposition rate ((18.37±3.14)%) and neuronal apoptosis rate ((27.58±2.63)%) in the inhibitor group were higher than those in the high-dose Dio group ((6.35±1.08)%, (14.02±1.87)%) (both P<0.05). (4) The levels of GSH, ROS, MDA, and Fe 2+ in the hippocampus of the 6 groups of rats showed statistically significant differences ( F=54.465, 106.453, 55.969, 105.457, all P<0.001). The GSH content in the inhibitor group ((103.48±8.76) mmol/g) was lower than that in the high-dose Dio group ((133.97±10.54) mmol/g), while the contents of ROS, MDA, Fe 2+ ((225.17±16.32) μmol/mg, (10.73±1.58) μmol/mg, (62.71±5.43) μg/g) were higher than those of the high-dose Dio group ((131.87±11.67) μmol/mg, (4.35±0.87) μmol/mg, (34.86±2.95) μg/g) (all P<0.05). (5)There were statistically significant differences in the protein levels of Tf, PCNA, Bax, caspase-3, SIRT1, Nrf2, HO-1 and Gpx4 in the hippocampus of the 6 groups of rats ( F=120.179, 107.568, 157.265, 98.031, 90.932, 52.283, 59.424, 114.539, all P<0.001). The protein levels of Tf, Bax and caspase-3 in the hippocampus of inhibitor group were higher than those of the high-dose Dio group, while the protein levels of PCNA, SIRT1, Nrf2, HO-1, Gpx4 were lower than those of the high-dose Dio group (all P<0.05). Conclusion:Diosmetin can activate SIRT1/Nrf2/HO-1/Gpx4 signaling pathway, thereby inhibiting neuronal ferroptosis in BM rats.

Diosmetin inhibits cell proliferation and promotes apoptosis through STAT3/c-Myc signaling pathway in human osteosarcoma cells

BACKGROUND: Diosmetin is a bioflavonoid compound naturally abundant in citrus fruits. It is found to perform a variety of activities, while its antitumor property in osteosarcoma, a malignant tumor with unmet clinical treatment, remained unknown. METHODS: Colony formation assay, cell cycle analysis and apoptosis analysis were conducted respectively to observe the effect of diosmetin on cell proliferation and apoptosis in human osteosarcoma cells. Western blot and immunoprecipitation were used to detect the expression of apoptotic molecules and activation of STAT3/c-Myc pathway in Saos-2 and U2SO cells. RESULTS: Diosmetin significantly inhibited cell proliferation, induced cell cycle arrest at G2/M phase and promoted cell apoptosis in both Saos-2 and U2SO cells. Moreover, Diosmetin downregulated the expression of anti-apoptotic protein Bcl-xL while upregulated the levels of pro-apoptotic proteins including cleaved Caspase-3, cleaved-PARP and Bax. Furthermore, diosmetin dose-dependently inhibited STAT3 phosphorylation, reduced the expression of its downstream protein c-Myc and impeded the interaction between STAT3 molecules. CONCLUSIONS: These results suggest that diosmetin exerts anti-osteosarcoma effects by suppressing cell proliferation and inducing apoptosis via inhibiting the activation of STAT3/c-Myc signaling pathway, which provide the possibility for diosmetin to be a chemotherapeutic candidate for osteosarcoma.

A new prenylated flavonoid glycoside from Sinopodophyllum hexandrum / 中草药

Objective: To study the chemical constituents from Sinopodophyllum hexandrum and their antitumor activities. Methods: The constituents were separated by chromatography of silica gel, ODS, Sephadex LH20 and pre-TLC. Their structures were elucidated by spectroscopic means. The in vitro cytotoxic activities of the isolated compounds were studied by MTT method. Results: Nine compounds were isolated and identified as 8,2'-diprenylquercetin 3-methyl ether-4'-O-β-D-glucoside (1), 8,2'-diprenyl quercetin-3- methylether (2), 5,7,4'-trihydroxy-3'-(3-methylbut-2-enyl)-3-methoxy flavone (3), 8-prenylkaempferol (4), sophoflavescenol (5), podoverine A (6), sinoflavonoid K (7), diosmetin (8) and acacetin (9). Conclusion: Compound 1 is a new compound named sinoflavonoid glycosides A, and compounds 5-9 are isolated from S. hexandrum for the first time. Compounds 1-5 show cytotoxicities against HeLa cells with IC50 of 42.6, 46.9, 26.9, 16.1 and 31.2 μmol/L, respectively.

Phytomolecules for Therapeutic Targets of Alzheimer’s Disease Pathology - A Virtual Analysis

Introduction: Life style of humans, with changes in diet, exercise and life style practices which play an important role in enhancing the progression of age related degenerative problems like dementia. The most common cause of dementia in the world is Alzheimer’s disease which ultimately decrease the cognitive function mainly learning and memory. The objective of the study was to find the multi target potential efficacy of the ligands, Glabridin and Diosmetin in altering the two main molecular targets of Alzheimer’s disease (AD). The target enzymes were amyloid binding alcohol dehydrogenase (ABAD) and β-site amyloid precursor protein cleaving enzyme 1(BACE1). Material and methods: In this study, we analyzed that multitarget potential of the two natural compounds on the two prior target enzymes of Alzheimer’s disease which are mainly involved in producing neurodegeneration. Drug likeness properties, absorption, digestion metabolic and toxicity profile and molecular docking were analyzed to determine therapeutic aspect by virtual methods. Results: Binding energy and Vander Waals force of Diosmetin were higher than Glabridin with the target ABAD and less than with BACE1 which showed that both drugs can be used in modulating the enzyme phosphorylation in Alzheimer’s disease. Conclusion: Glabridin and Diosmetin could be used as promising drug candidates as ABAD inhibitor and BACE1 inhibitor in Alzheimer’s disease

Protective effect of diosmetin on viral myocarditis of CVB3-induced suckling mice by regulating macrophage M1/M2 type polarization / 中国药理学与毒理学杂志

Objective: To study the protective effect of diosmetin on viral myocarditis induced by coxsackie virus B3 (CVB3) by regulating macrophage M1/M2 type polarization. Methods: BALB/c suckling mice were given ip 0.1 mL CVB3 virus culture solution of 1×105pfu, which was recorded as DO. After 24 h (D1), diosmetin (Dio) 12.5, 25 and 50 mg·kg-1were ig given, respectively, 7 d in succession. At the end of the experiment, mean arterial pressure (MAP), heart rate (HR), left ventricular systolic pressure (LVSP), expression levels of serum creatine kinase (CK), creatine kinase MB isoenzyme (CK-MB) and myoglobin (Mb) were detected. The peripheral blood was collected to analyze and sort polarization of macrophage M1 and M2 in peripheral blood by flow cytometry. The pathological features of myocardial tissues were analyzed by HE staining and TUNEL staining. Western blotting was performed to detect the expressions of cleaved-caspase 3 and cleaved-caspase 9 in myocardial tissues. Macrophages were sorted from spleen tissues of suckling mice. After macrophages were treated with IFN-y 5 mg·L-1, CVB3 1×105pfu, Dio 20 μmol·L-1, CVB3 (1×105pfu)+Dio 20 μmol·L-1for 24 h, the expression levels of inducible nitric oxide synthase (iNOS) and arginase-1 (Arg-1) in cells were detected by Western blotting. Results: Compared with normal control group, MAP, HR and LVSP in model group were significantly decreased (P<0.05). Compared with model group, MAP, HR and LVSP were significantly increased in Dio 12.5, 25 and 50 mg·kg-1groups (P<0.05). Compared with normal control group, levels of CK, CK-MB and Mb in serum, and apoptosis rate of cardiomyocytes in model group were significantly increased (P<0.05), so were expression levels of cleaved-caspase 3 and cleaved-caspase 9 in myocardial tissues (P<0.05). Compared with model group, levels of CK, CK-MB and Mb in serum, and apoptosis rate of cardiomyocytes were significantly decreased in Dio 12.5, 25 and 50 mg·kg-1groups (P<0.05), so were expression levels of cleaved-caspase 3 and cleaved-caspase 9 in myocardial tissues (P< 0.05). Compared with normal control group, positive percentages of F4/80+iNOS\F4/80+TLR4+and F4/ 80+CD40+in peripheral blood of model group were significantly increased, while those of F4/80+CD14\ F4/80+Arg-1+ and F4/80+CD206t were significantly decreased (P<0.05). Compared with model group, the former three items in peripheral blood were significantly decreased in Dio 12.5, 25 and 50 mg·kg-1groups, while the latter three items were significantly increased (P<0.05). Both IFN-γ and CVB3 virus could up-regulate the expression of iNOS, and inhibit the expression of Arg-1. The Dio treatment could inhibit the expression of iNOS and up- regulate the expression of Arg-1. Compared with CVB3 virus alone group, the expression of iNOS was decreased, but the expression of Arg-1 was increased in CVB3 + Dio group (P<0.05). Conclusion: Dio can inhibit the activation of M1 type macrophage caused by viral infection, promote its development towards M2 polarization, and improve cardiac function of the viral myocarditis model.

Diosmetin Alleviates Lipopolysaccharide-Induced Acute Lung Injury through Activating the Nrf2 Pathway and Inhibiting the NLRP3 Inflammasome

Acute lung injury (ALI)/acute respiratory distress syndrome (ARDS) is a common clinical syndrome of diffuse lung inflammation with high mortality rates and limited therapeutic methods. Diosmetin, an active component from Chinese herbs, has long been noticed because of its antioxidant and anti-inflammatory activities. The aim of this study was to evaluate the effects of diosmetin on LPS-induced ALI model and unveil the possible mechanisms. Our results revealed that pretreatment with diosmetin effectively alleviated lung histopathological changes, which were further evaluated by lung injury scores. Diosmetin also decreased lung wet/dry ratios, as well as total protein levels, inflammatory cell infiltration and proinflammatory cytokine (eg. TNF-α, IL-1β and IL-6) overproduction in bronchoalveolar lavage fluid (BALF). Additionally, increased MPO, MDA and ROS levels induced by LPS were also markly suppressed by diosmetin. Furthermore, diosmetin significantly increased the expression of Nrf2 along with its target gene HO-1 and blocked the activation of NLRP3 inflammasome in the lung tissues, which might be central to the protective effects of diosmetin. Further supporting these results, in vitro experiments also showed that diosmetin activated Nrf2 and HO-1, as well as inhibited the NLRP3 inflammasome in both RAW264.7 and A549 cells. The present study highlights the protective effects of diosmetin on LPS-induced ALI via activation of Nrf2 and inhibition of NLRP3 inflammasome, bringing up the hope of its application as a therapeutic drug towards LPS-induced ALI.

Flavonoids from the flowers of Chrysanthemum morifolium Ramat / 中国药学杂志

OBJECTIVE: To study the chemical constituents in the flowers of Chrysanthemum morifolium Ramat. METHODS: The compounds were isolated with Diaion HP-20, Toyopearl HW-40, Sephadex LH-20, silica gel column chromatography and preparative HPLC. The structures of the compounds were identified by physiochemical properties and spectral analysis. RESULTS: Twenty compounds were obtained, and their structures were identified as luteolin (1), apigenin (2), acacetin (3), diosmetin (4), acacetin 7-O-β-D-glucoside (5), acacetin 7-O-β-D-glucoside (6), acacetin7-O-(6″-O-acetyl)-β-D-glucoside (7), eriodictyol (8), naringenin (9), artemetin (10), 5-hydroxy-6, 7, 3', 4'-tetramethoxyflavone (11), 5, 7-dihydroxy-3', 4'-dimethoxyflavone (12), 4'-methoxyctricin (13), 3', 5'-dimethoxy-4', 5, 7-trihydroxyflavone (14), 5, 6-dihydroxy-3, 7, 3', 4'-tetramethoxyflavone (15), luteolin 7-O-β-D-glucuronide methyle ester (16), dihydroquercetin-7-β-D-glucoside (17), quercetin 3-O-β-D-glucoside(18), quercetin 3-O-β-D-glucoside (19), and acacetin 7-O-β-(6″-(E)-crotonylglucopyranoside) (20). CONCLUSION: Compounds 9-20 were isolated for the first time from this plant.

Determination of six flavonoids in Schizonepeta tenuifolia from different areas by HPLC / 中草药

Objective: To detect the contents of six kinds of flavonoids in Schizonepeta tenuifolia from different areas, so as to provide a basis for their quality control. Methods: Ultra performance liquid chromatography (UPLC) was used to establish the detection scheme of flavonoids content, and clustering analysis was conducted by the results of content detection. Results: The contents of six kinds of flavonoids in S. tenuifolia from different areas had large differences. The linear ranges were: 0.030-0.148 μg for cynaroside, 0.222-1.108 μg for quercetin, 0.357-1.784 μg for hesperidin, 0.058-0.292 μg for luteolin, 0.054-0.269 μg for apigenin, and 0.050-0.247 μg for diosmetin, respectively. The average recovery rate was between 96.32% and 99.97%, RSD < 2.20%; Among this, the Schizonepeta Briq. herb from Anhui was as one class, the herbs from Henan 1 and Guangdong were as one class, and the herbs from Henan 2, Hebei, Yunnan, Sichuan, Hubei, and Gansu were as the other class. Conclusion: The method used for quality control of the multi-index level in this experiment is simple and sensitive; on the other hand, the herbs from Henan 1, Henan 2, Hebei, Yunnan, Sichuan, Hubei, Guangdong and Gansu have a close relationship; They can be used as the same medicinal source.

Simultaneous determination of five effective components in different parts of Dracocephalum moldevica L by RP-HPLC / 中国药学杂志

OBJECTIVE: To develop an RP-HPLC method for simultaneous determination of rosmarinic acid, tilianin, luteolin-7-O-β-D-glucuronide, apigenin-7-O-β-D-glucuronide, and diosmetin-7-O-β-D-glucuronide in different parts ofDracocephalum moldevica L. METHODS: The five constituents were measured on a Shim-pack ODS column(4.6 mm×250 mm, 5 μm)with gradient elution of acetonitrile (A)-0.5% formic acid aqeous solution (B) (0-30 min, 17%A;30-60 min, 17%-28%A; 60-70 min, 28%A) at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 330 nm, and the column temperature was maitained at 35℃. RESULTS: The linear ranges of rosmarinic acid, tilianin, luteolin-7-O-β-D-glucuronide, apigenin-7-O-β-D-glucuronide, and diosmetin-7-O-β-D-glucuronide were 4.2-126 μg·mL-1 (r=0.999 2), 7.84-235.2 μg·mL-1 (r=0.999 3), 3.048-91.44 μg·mL-1(r=0.999 4), 1.472-44.16 μg·mL-1(r=0.999 4), and 2.816-84.48 μg·mL-1 (r=0.999 2), respectively. The average recoveries (RSD) of the five compounds were 98.97%(1.03%), 99.90%(0.92%), 99.89% (1.75%), 99.55% (0.98%), and 99.76%(1.19%) (n=6), respectively. CONCLUSION: The developed method is accurate and precise, which can be used for the quality control of different parts of Dracocephalum moldevica L.The RESULTS: of content determination indicate that the five compounds exist in all the parts of Dracocephalum moldevica L., but the mass fractions are obviously different.

Separation of flavonoids from stems and leaves of male plants in Trichosanthes kirilowii and study on their activity of scavenging DPPH free radical / 中草药

Objective: To study the flavonoids of the stems and leaves of male Trichosanthes kirilowii and to make a primary research on the structure activity relationship between flavonoids and their DPPH-scavenging capacity. Methods: Flavonoids from the stems and leaves of male T. kirilowii were separated by chromatographic techniques, such as polyamide resin, high-speed countercurrent chromatography and high performance liquid chromatography. According to the chemical properties and spectral analysis, the chemical structures of the compounds were identified. And we determined the antioxidant ability in vitro of seven flavonoids by DPPH methods. Results: Seven flavonoids were isolated from the stems and leaves of male T. kirilowii. They were luteolin (1), chrysoeriol (2), luteolin-7-O-β-D-glucoside (3), chrysoeriol-7-O-β-D-glucoside (4), apigenin-7-O-β-D-glucoside (5), diosmetin-7-O-β-D-glucoside (6), and quercetin-3-O-β-glucoside (7). Under the present experimental condition, the order of their DPPH-scavenging capacities was 1 > 3 > 7 > 2 > 4> 6 > 5. Conclusion: Compounds 1, 2, 6, and 7 are isolated from this part of male T. kirilowii for the first time. DPPH-scavenging capacities of the compounds 1, 3, and 7 are much stronger than others, but they all have 3',4' two adjacent hydroxide groups in B ring on the view of structure. DPPH-scavenging capacities of compounds 4 and 6 are much weaker than compound 3, but the former have 3' or 4' hydroxyl methylation in the structure. DPPH-scavenging capacity will also decrease if there is a 7 hydroxyl glycosylation in ring A by comparing compound 1 and 3. We speculate that it is caused by the increase of the steric hindrance.

Flavonoid profile and antioxidant activities of methanolic extract of Hyparrhenia hirta (L.) Stapf.

In this study, we report isolation of flavonoids, viz., 3-O-methylquercetin, tangeritin, luteolin-7-O-glucoside, luteolin, apigenin-7-O-glucoside, apigenin-8-C-glucoside, luteolin-8-C-glucoside, luteolin-6-C-glucoside, diosmetin and catechin from the methanolic extract of Hyparrhenia hirta employing high performance liquid chromatography and liquid chromatography-electrospray ionization-tandem mass spectrometry. The total phenolic content of H. hirta extract was 105.58 ± 0.1 mg gallic acid equivalents/g of plant extract while the total flavonoid content was 45.20 ± 0.2 mg quercetin equivalents/g of plant extract and the total condensed tannin were 72.35 ± 0.7 mg catechin equivalents/g of plant extract by reference to standard curve. The antioxidant activity was assayed through the antioxidant capacity by phosphomolybdenum assay, the reducing power assay and the radical scavenging activity using 2,2-diphenyl-1-picrylhydrazyl method. The extract showed dose dependant activity in all the three assays.

Extraction technology for active constituents in Dracocephali Moldavici Herba and comparison on contents of Dracocephali Moldavici Herba from various habitats / 中草药

To study the extraction technology for the active constituents in Dracocephali Moldavici Herba (the aerial parts of Dracocephalum moldevica) and to compare their contents Dracocephali Moldavici Herba from various habitats. The contents of luteolin-7-O-glucuronide (I), apigenin-7-O-glucuronide (II), rosmarinic acid, diosmetin-7-O-glucuronide (III), tilianin, and acacetin-7-O-glucuronide (IV) in Dracocephali Moldavici Herba were measured using HPLC method. Based on single-factor test, and the influence of the extracting menstrua, menstruum dosage, and extracting time were investigated using orthogonal design method. Based on the optimal technology, the contents of I, II, rosmarinic acid, III, tilianin, and IV in Dracocephali Moldavici Herba from various habitats were compared. The optimal conditions for the extracting for one time, each time for 5 h with 50 times of amount of 40% ethanol. D. moldevica produced in Jimusar, Xinjiang had the higher contents of I, II, rosmarinic acid, III, tilianin, and acacetin-7-O-glucuronide. This extraction technology is reasonable, stable, and feasible. The contents of I, II, rosmarinic acid, III, tilianin, and IV in Dracocephali Moldavici Herba from different habitats have some differences. The multi-index contents of I, apigenin-7-O-glucuronide, rosmarinic acid, III, tilianin, and IV could reflect the quality of Dracocephali Moldavici Herba more comprehensively.

Chemical constituents from whole herb of Lagochilus platyacanthus / 中草药

Objective: To study the chemical constituents from the whole herb of Lagochilus platyacanthus. Methods: The chemical constituents were isolated and purified by various chromatographic techniques and the structures were identified by spectral analysis. Results: Twenty-one compounds were isolated from the 95% ethanol extract of L. platyacanthus, including 15 flavonoids: apigenin-7, 4'-dimehyl ether (1), acacetin (2), apigenin (3), luteolin-7, 3', 4'-trimethyl ether (4), luteolin-7, 4'-dimethyl ether (5), diosmetin (6), chrysoeriol (7), quercetin-3-O-rutinoside-7-O-glucoside (8), rutin (9), horridin (10), apigenin-6, 8-di-C-β-D-glucopyranoside (11), isorhamnetin-3-O-rutinoside (12), isorhamnetin-3-O-robinobioside (13), isorhamnetin-3-O-β-D-glucoside (14), and isorhamnetin-3-O- rutinoside-4'-O-glucoside (15); three lignans: 1-(4-hydroxy-3-methoxy)-phenyl-2-[4-(1, 2, 3-trihydroxypropyl)-2-methoxy]-phenoxy- 1, 3-propandiol (16), (+)-isolarisiresinol 3-α-O-β-D-glucopyranoside (17), and (-)-isolarisiresinol 3-α-O-β-D-glucopyranoside (18); two iridoids: 8-O-acetylharpagide (19) and geniposidic acid (20), and one phenylethanoid glycoside: lavandulifolioside (21). Conclusion: All the compounds are obtained from this plant for the first time. The compounds are isolated from the plants in genus Lagochilus Bunge for the first time except compounds 1, 3, 9, 19, and 20.

Chemical constituents from Artemisiae Anomalae Herba / 中草药

Objective: To investigate the chemical constituents from Artemisiae Anomalae Herba (the aerial part of Artemisia anomala or A. actiflora). Methods: The chemical constituents from Artemisiae Anomalae Herba were isolated and purified by chromatography on silica gel and Sephadex LH-20 column, as well as preparative HPLC. Their structures were identified on the base of physicochemical properties and spectroscopic data analyses. Results: Fifteen constituents, including seven steroids and eight flavonoids, were isolated and identified as β-sitosterol (1), β-daucosterol (2), schleicheol 2 (3), α-spinasterol (4), 5α, 8α-epidioxy- ergosta-6, 22-dien-3β-ol (5), 5α, 8α-epidioxy-ergosta-6, 9(11), 22- trien-3β-ol (6), 22E-3β, 5α-dihydroxyergosta-7, 22-dien-6-one (7), naringenin (8), luteolin (9), kaempferol (10), chrysoeriol (11), diosmetin (12), jaceosidin (13), isorhamnetin 3-O-glucoside (14), and hesperetin-7-O-β-D-glucopyranoside (15). Conclusion: Compounds 5, 6, 12-14 are isolated from this plant for the first time, compound 3 is isolated from the plants of Artemisia L. for the first time, and compounds 7 and 15 are isolated from the plants in Compositae family for the first time.

Mechanism of flavonoids from Galium verum on cell proliferation and apoptosis of HepG2 / 中草药

Objective: To investigate the inhibition of cell proliferation and induction of apoptosis of flavonoids [diosmetin-7-O-β-D-xylopyranosyl-(1 → 6)-β-D-glucopyranoside, DXG)] from Gallium verum on liver cancer cell HepG2 and to study its mechanisms. Methods: Cell growth was detected with Trypan blue staining. The proliferation inhibition rates of DXG on HepG2 were measured by MTT assay. The morphology changes of apoptotic cell were obserbed by AO-EB double staining. Agarose gel electrophoresis was used to detect the DNA fragmentation. The expression of apoptosis-related genes bax and bcl-2 was examined by RT-PCR. Results: DXG (50, 100, and 200 μg/mL) could significantly inhibit the proliferation of HepG2 cells by Trypan blue staining and MTT assay compared with the control group (P < 0.05). AO-EB double staining showed that DXG could induce the apoptosis and morphological change of HepG2 cells and the ratios of apoptosis were obviously high when the concentration of DXG increased. The laddering pattern was clearly presented in the cells treated with 100 and 200 μg/mL DXG for 48 h. RT-PCR showed that DXG could up-regulate the mRNA level of bax and down-regulate the mRNA levels of bcl-2 in HepG2 cells. Conclusion: DXG could obviously inhibit the proliferation and induce the apoptosis in HepG2 cells. The mechanism is related to the modulation of the expression level of bax/bcl-2 mRNA.