Improvement effects of aucubin on ADHD-like behaviors in mice via the inhibition of excessive astrocytic activation / 中华行为医学与脑科学杂志

Objective:To investigate the effect of aucubin on behaviors and excessive activation of astrocytic in attention deficit/hyperactivity disorder (ADHD) model mice.Methods:Twelve wild-type C57BL/6 pregnant mice (female, clean grade) were intraperitoneally administered with esketamine (15 mg/kg) to establish an ADHD model in offspring mice. The offspring mice were divided into control+ saline group, control+ aucubin group, Ketamine+ saline group and Ketamine+ aucubin group according to the nest matching principle with 15 in each group.At 14 days after birth, mice in the control+ aucubin group and Ketamine+ aucubin group were administered with aucubin (5 mg/kg, once a day) by gavage for 5 days. Mice in control+ saline group and Ketamine+ saline group were administered with equal volume of 0.9% sodium chloride solution. The offspring mice were housed with their mothers in the same cage until 21 days after birth. Twenty-one days after birth, the offspring mice were evaluated by open field test and elevated plus maze tests. Immunofluorescence assay was used to detect the expression of glutamate decarboxylase 2 (GAD2), γ- aminobutyric acid (GABA) and glial fibrillary acidic protein (GFAP) in the amygdala. The morphological changes of astrocytes were quantitatively analyzed by Sholl analysis. GraphPad Prim 9.0.1 software was used for statistical analysis. The comparison of multiple groups was conducted by one-way ANOVA or Kruskal-Wallis test.Results:(1)The results of behavioral experiments showed that the total distance traveled in the open field test and the residence time in open arm of the elevated plus maze were statistically significant ( F=236.90, H=39.92, both P<0.001). The total distance ((7 044±249)mm, (22 891±2 175)mm, P<0.05) and the residence time in open arm(12.69(9.86, 17.24)s, 2.72(0.57, 3.87)s, P<0.05) of mice in Ketamine+ saline group were both higher than those in control+ saline group.The total distance((22 891±2 175)mm, (8 252±839)mm, P<0.05) and the the residence time in open arm(5.45(1.13, 10.99)s, 12.69(9.86, 17.24)s, P<0.05) of Ketamine+ aucubin group were both lower than those of Ketamine+ saline group.(2)The immunofluorescence results showed that the levels of GAD2, GABA and GFAP intensity in amygdala of mice in the four groups were statistically significant ( F=145.50, 50.08, 53.83, all P<0.05). Compared with control+ saline group, the fluorescence intensities of GAD2 ((100.00±9.60)%, (24.86±4.14)%, P<0.05) and GABA ((100.00±16.84))%, (25.48±5.70)%, P<0.05) of Ketamine+ saline group were down-regulated, and the GFAP((100.00±18.02)%, (223.80±25.85)%, P<0.05) was up-regulated. Compared with Ketamine+ saline group, the fluorescence intensities of GAD2 ((24.86±4.14)%, (56.08±6.55)%, P<0.05) and GABA((25.48±5.70)%, (52.59±15.74)%, P<0.05) in Ketamine+ aucubin group were up-regulated, but the fluorescence intensity of GFAP ((223.80±25.85)%, (157.10±22.10)%, P<0.05) was down-regulated.(3)Sholl analysis indicated that the number of the intersections between the astrocyte processes or the branches of astrocyte processes was statistically significant in the 4 groups ( F=12.47, P<0.05). Compared with control+ saline group, the number of the intersections in Ketamine+ saline group((2.07±0.48), (1.67±0.72), P<0.05) increased. While the number of the intersections in Ketamine+ aucubin group was lower than that of Ketamine+ saline group ((1.20±0.78), (2.07±0.48), P<0.05). Conclusion:Aucubin administration can alleviate ADHD-like behaviors in offspring mice, and the mechanism may be associated with the inhibition of excessive astrocytic activation.

Anti-inflammatory effects of aucubin in cellular and animal models of rheumatoid arthritis / 中国天然药物

Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease. It is known that aucubin (AU) exerts anti-inflammatory activity, but its effects and mechanisms in RA are unclear. This study investigated the anti-inflammatory effects and mechanisms of AU in vivo and in vitro. Human fibroblast-like synoviocyte cells from patients with RA (HFLS-RA), RAW264.7 cells, and MC3T3-E1 cells were used to evaluate the effects of AU on migration, invasion, apoptosis, osteoclast differentiation and production. Immunofluorescence was used to observe nuclear translocation of nuclear factor (NF)-κB, the double luciferase reporter gene method was used to observe NF-κB-p65 activity in AU-treated MC3T3-E1 cells. RT-qPCR was used to measure expression of bone metabolism and inflammation-related genes, and western blot was used to measure bone metabolism and NF-κB protein expression levels. Collagen-induced arthritis (CIA) rat model was used for pharmacodynamics study. Arthritis indexes were measured in the ankle and knee, histological staining and Micro-computed tomography were performed on the ankle joints. Also, inflammatory factor gene expression and the levels of NF-κB-related proteins were detected as in vitro. AU effectively inhibited HFLS-RA cell migration and invasion, promoted apoptosis, and inhibited RAW264.7 cell differentiation into osteoclasts, as well as inhibited NF-κB-p65 activity in MC3T3-E1 cells. Notably, AU significantly reduced the gene expression levels of three cell-related inflammatory factors and bone metabolism factors, effectively inhibited the expression of p-Iκκα β, p-IκBα, and p-p65 proteins. In vivo, AU relieved joint inflammation, reduced related inflammatory factors, and inhibited NF-κB signaling. It could be used to treat RA-related synovial inflammation and bone destruction through the NF-κB pathway.

Dissolution rule of Siwu Decoction by different decocting methods based on chemistry holography / 中草药

Objective: To study the content variation and chemical composition of Siwu Decoction between mixed decoction and single decoction comprehensively, and then explore variation rule of Siwu Decoction by different decocting methods based on material basis. Methods: Components of Siwu Decoction were identified by LC-MS/MS and an UPLC wavelength switching method for simultaneously determining the contents of multiple compounds in Siwu Decoction was established based on the idea of TCM chemistry holography. The mixed and single decoction samples were prepared and tested. Experimental data was compared to analyze material basis differences and variation rule of Siwu Decoction by different decocting methods. Results: A total of 72 compounds were identified and assigned, 18 compounds were quantitative detected and all of 18 analytes showed good linearity (R2 ≥ 0.999) within the test range. The relative standard deviations of the precision, repeatability and stability were not exceeding 2.0%, and the recoveries were in the range of 97%-105%. Analysis of Siwu Decoction samples showed dissolution of ligustilide, 3-n- butylphthalide, catechin, gallic acid and paeoniflorin was affected by the change of solvent volume and dissolution of aucubin, catechin, oxypaeoniflorin, paeoniflorin and acteoside were higher in mixed decoction than single decoction obviously. Compared to single decoction, the kinds of compounds in mixed decoction did not change significantly but the content showed notable variety. Conclusion: Through the study of chemistry holography, the composition and content of compounds in TCM mixed decoction and herbs single decoction can be compared and analyzed comprehensively to provide a new perspective for the study on the rule of TCM decoction and dissolution. TCM chemistry holography study may become a useful exploration of the TCM quality study.

Determination, Isolation, And Identification Of Aucubin And Verbascoside In The Leaves Of Iraqi Plantago Lancoleta L. Using Different Detecting Methods

Objective: Plantago lanceoleta L. (ribwort plantain) is one of the important medicinal herbs which is widespread fortune available in Iraq, that have a wide range of medicinal properties. The aim of this work was to determine, isolate and identify verbascoside and aucubin in Iraqi P. lanceoleta L. by using different chromatographic and spectrometric methods. Methods: Verbascoside and aucubin were isolated and quantified by preparative TLC, and then they were determined by the high-performance thin-layer chromatography (HPTLC) fingerprinting. Aucubin and catalpol in the plant extract were analyzed by liquid chromatography-mass spectrometry (LC-MS); aucubin and verbascoside that isolated from the plant sample were examined by fourier-transform infrared spectroscopy (FT-IR) and LC-MS, respectively. Results: The result showed that the Iraqi P. lanceoleta L. contains 1.74 percent (verbascoside) and 0.24 percent (aucubin) of dry powdered leaves. Each TLC-isolated compound showed a single spot on the HPTLC plate, which give an idea about the purity of the isolated compound. Aucubin (with catalpol) and verbascoside both are detected by LC-MS in different ionization mode. Many functional groups were identified in the TLC-isolated aucubin by FT-IR. Conclusion: The Iraqi P. lanceoleta L. showed a high content of verbasoside, and it is a very rich source for this compound, which can be easily isolated by TLC and subjected to many pharmacological studies. The extract of the young leaves of this plant gave a little amount of aucubin, and it is easy to obtain a higher content from the older leaves.

Comparison of Catalpol and Aucubin Contents in Different Parts of Wild Centranthera grandiflora / 中国药房

OBJECTIVE： To determine and compare the contents of catalpol and aucubin in different parts （root， stem， leaf and flower） of wild Centranthera grandiflora， and to provide reference for the selection of medicinal parts and source development. METHODS： HPLC method was used to determine the contents of catalpol and aucubin in root， stem， leaf and flower of wild C. grandiflora， and the contents of different parts were analyzed comparatively. The determination of catalpol was performed on Agilent TC-C18 column with mobile phase consisted of methanol-0.1％ phosphoric acid （1 ∶ 99， V/V） at the flow rate of 1 mL/min； the detection wavelength was set at 210 nm， and sample size was 20 μL. The column temperature was 35 ℃； the determination of aucubin was performed on SPHERI-5RP-C18 column with mobile phase consisted of acetonitrile-water （3 ∶ 97， V/V） at the flow rate of 1 mL/min； the detection wavelength was set at 205 nm， and sample size was 20 μL； the column temperature was 25 ℃. RESULTS： The linear range of catalpol and aucubin were 0.061 5-3.321 and 0.000 36-0.216 mg/mL （all r＝0.999 9）. The limits of detection were 0.016 and 0.007 μg/mL. The limits of quantitation were 0.052 and 0.023 μg/mL. RSDs of precision， stability （24 h） and reproducibility tests were all lower than 2.00％ （n＝6）. The average recoveries were 99.34％ and 99.61％， and RSDs were 1.06％ and 1.12％， respectively （n＝6）. The average content of catalpol in root， stem， leaf and flower wild C. grandiflora were 1.609， 3.030， 11.095 and 1.921 mg/g， respectively. The contents of aucubin in different parts were 0.441， 0.020， 0.005 and 0.006 mg/g，respectively. CONCLUSIONS：The established HPLC method meets the requirements of quantitative analysis. Catalpol is mainly distributed in the leaves of wild C. grandiflora， and aucubin is mainly distributed in the roots of wild C. grandiflora. The experimental conclusion provides a reference for the reasonable selection of different medicinal parts as raw materials to develop medicine with different efficacy.

Simultaneous Determination of Contents of 8 Components in Eucommia ulmoides Leaves by HPLC / 中国药房

OBJECTIVE： To establish content determination method for simultaneously determining aucubin，geniposidic acid，catechin，chlorogenic acid，asperuloside，rutin，isoquercitrin and astragalin in Eucommia ulmoides leaves. METHODS：HPLC method was adopted. The determination was performed on Agilent ZORBAX SB-C18 column with the mobile phase consisted of acetonitrile-0.1％ phosphoric acid（gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 203 nm for aucubin and catechin，239 nm for geniposidic acid and asperuloside，220 nm for chlorogenic acid，354 nm for rutin and isoquercitrin，and 266 nm for astragalin. The sample size was 5 μL. RESULTS： The linear range of aucubin， geniposidic acid， catechin， chlorogenic acid， asperuloside， rutin， isoquercitrin and astragalin were 0.812-6.090 μg（r＝0.999 3），0.438-3.285 μg（r＝0.999 2），0.045-0.336 μg（r＝0.999 2），0.882-6.615 μg（r＝0.999 3），0.097-0.726 μg（r＝0.999 1），0.064-0.483 μg（r＝0.999 3），0.048-0.360 μg（r＝0.999 1） and 0.014-0.108 μg（r＝0.999 7），respectively. RSDs of precision， stability and reproducibility tests were all lower than 3.5％（n＝6）. The average recovery rates were 101.60％，103.06％，99.77％，96.93％，98.17％，96.75％，98.97％ and 99.60％，with RSDs of 1.42％，2.65％，2.78％，2.05％，2.26％，0.93％，2.79％ and 3.08％，respectively（n＝6）. The contents of aucubin， geniposide， catechin， chlorogenic acid， plantain， rutin， isoquercetin and astragaloside in 12 batches of E. ulmoides leaves from different collection time and planting varieties were 10.903-17.245， 5.578-7.892， 0.198-0.440， 13.890-19.782， 1.008-1.547， 1.102-2.396， 0.267-0.701， 0.150-0.412 mg/g， respectively. The content fluctuated greatly. The contents of aucubin， geniposide， catechin， chlorogenic acid， rutin and pinoresinol diglucoside in cortex of E. ulmoides were 0.299， 0.123， 0.580， 0.112， 0.026，1.961 mg/g， respectively. CONCLUSIONS： The method is simple， reproducible and accurate. It can be used to evaluate the quality of E. ulmoides leaves. There are obvious differences in composition and content of components in different medicinal parts （cortex， leaves） of E. ulmoides.

Optimization of steaming technology for decoction pieces of Scrophularia ningpoensis by D-optimal response surface methodology based on AHP- entropy weight method / 中草药

Objective :To determine the weight coefficient and optimize the steaming technology of decoction pieces of Scrophularia ningpoensis (SN). Methods The contents of harpagide, harpagoside, aucubin, acteoside, angoroside C, and cinnamic acid were simultaneously determined by HPLC. The weight coefficient of each component was evaluated by AHP-entropy (analyitc hierarchy process) method. With composite score as index, D-optimal response surface methodology was adopted to investigate the effects of soaking time, steaming time, and drying temperature on the quality of processed products and optimize the processing technology of decoction pieces of SN. Results :Optimal processing parameters were as follows: soaking time was 15.63 min, steaming time was 85 min, drying temperature was 60 ℃, and the synthetical mark was 97.20. Considering the actual situation, the optimum processing technology of SN was obtained by fine-turning the soaking time. The soaking time was 15 min, the steaming time was 85 min, and the drying temperature was 60 ℃. Also, the synthetical mark were 98.53, 99.39, 98.86, and its RSD was 0.47% through the obtained conditions in parallel with three batches of samples. Conclusion: The optimized steaming technology is simple and feasible, which can provide a reference for the steaming of decoction pieces of SN. The method established to simultaneously determine the contents of six components in decoction pieces of SN is rapid and reliable for controlling the quality of decoction pieces of SN.

Protective effect of aucubinon on the eucommia ulmoides on skin photoaging induced by UVB / 国际中医中药杂志

Objective To explore the protective effect of aucubin on the photoageing of the skin that induced by ultraviolet (UVB) radiation. Methods Human skin fibroblasts were divided into normal group, model group and high, medium and low dose groups of apocynin by random method. Except the normal group, the other groups of cells were subjected to UVB radiation with a radiation wavelength of 311 nm, irradiation intensity of 9.00 mw/cm2, radiation time of 2.8 s, radiation dose of 25 mJ/cm2. The cells in the high, medium and low doses of aglycone were cultured for 24 h at an equal volume of 400, 200, 100 μg/ml, and then subjected to UVB irradiation, and then cultured for 24 h. The normal group and the model group were cultured for 48 h in a common medium. The apoptosis rate of each group was detected by flow cytometry. The levels of IL-1, IL-6 and TNF-α in each group were detected by ELISA. The expressions of Bax and Bcl-2 mRNA in each group were detected by RT-PCR. Results Compared with the model group, the apoptosis rate (4.84% ± 0.24%, 12.32% ± 0.67% vs. 35.63% ± 2.77%) in the medium and high dose group significantly decreased (P<0.05), the content of IL-1 (21.14 ± 1.94 ng/ml, 16.17 ± 0.74 ng/ml vs. 22.55 ± 1.02 ng/ml), IL-6 (17.46 ± 0.93 ng/ml, 14.51 ± 0.79 ng/ml vs. 18.39 ± 2.05 ng/ml), TNF-α (44.21 ± 1.16 ng/ml, 35.94 ± 3.08 ng/ml vs. 45.67 ± 2.28 ng/ml) significantly increased (P<0.05). Compared with the model group, the expression of Bax mRNA (0.29 ± 0.05, 0.42 ± 0.05, 0.51 ± 0.04 vs. 0.59 ± 0.05) significantly decreased, and the expression of Bcl-2 mRNA (0.52 ± 0.04, 0.44 ± 0.03, 0.35 ± 0.03 vs. 0.26 ± 0.04) significantly increased (P<0.05). Conclusions The aucubin has protective function to the damage of skin fibroblasts which is induced by UVB radiation, indicating the protective function of decreasing Bax mRNA expression, increasing Bcl-2 mRNA expression, reducing the expression of inflammatory factor IL-1, IL-6, TNF-α, holding-up the cell apoptosis which is induced by UVB radiation, and inhibiting apoptosis of cell that caused by UVB radiation, playing an role of protective fibroblasts for delay of the photoageing.

Aucubin regulates skin damage repair by TGF-β/P38MAPK signaling pathway / 国际中医中药杂志

Objective To explore the mechanism of transforming growth factor-beta/p38 mitogen-activated protein kinases (TGF-β/P38MAPK) signaling pathway that transformed corallin into a transforming growth factor-beta/p38 mitogen activated protein kinases (TGF-β/P38MAPK).Methods The hair follicle stem cells (HFSCs) were divided into blank group,positive group and 10-10,10-9,10-8,10-7,10-6,10-5,10-4,10-3 mol/L mycoralin solution groups according to the random number table method.After 24 h,the cell proliferation rate was determined by MTT assay.The HFSCs were divided into blank group,positive group and low,medium and high dose groups according to the random number table method.The positive group was added with 5 mol/L of alkaline fibroblast growth factor solution,while the low,medium and high dose groups were added with 10-7,10-6 and 10-5 mol/L of mycoralin solution for intervention.The cell proliferation rate was determined by MTT assay.The protein expressions ofTGF-β,phosphorylated P38(p-p38),P38,Bax and bcl-2 in cells were detected by Western blot,and mRNA expressions of Bax and bcl-2 in cells of each group were detected by real-time quantitative PCR.Results Compared with the blank group,the productivity of the cells (0.418 ± 0.031,0.412 ± 0.014,0.468 ± 0.024 vs.0.353 ± 0.006) in the 10-7,10-6 and 10-5 mol/L aucubin groups significantly increased (P＜0.01).Compared with the blank group,the protein expression of TGF-β (0.377 ± 0.027,0.338 ± 0.021,0.322 ± 0.017 vs.0.557 ± 0.017),p-p38 (0.270 ± 0.020,0.228 ± 0.013,0.216 ± 0.012 vs.0.461 ± 0.012),Bax (0.450 ± 0.017,0.365 ± 0.011,0.279 ± 0.006 vs.0.551 ± 0.015) in the low,medium and high dose groups significantly down-regulated,while the Bcl-2 (0.450 ± 0.017,0.365 ± 0.011,0.279 ± 0.006 vs.0.358 ± 0.011) protein expression significantly increased.Compared with the blank group,the expression of Bcl-2 (1.714 ± 0.028,2.514 ± 0.054,3.382 ± 0.084 vs.1.000 ± 0) mRNA increased,Bax (0.415 ± 0.020,0.353 ± 0.090,0.235 ± 0.114 vs.1.000 ± 0) mRNA in the low,medium and high dose groups significantly down-regulated (P＜0.01).Conclusions By regulating the TGF-β/p38MAPK signaling pathway,mycoralin can reduce the expression of downstream apoptotic factors,and promote the repair of damaged skin.

Aucubin Promotes Differentiation of Neural Precursor Cells into GABAergic Neurons

Aucubin is a small compound naturally found in traditional medicinal herbs with primarily anti-inflammatory and protective effects. In the nervous system, aucubin is reported to be neuroprotective by enhancing neuronal survival and inhibiting apoptotic cell death in cultures and disease models. Our previous data, however, suggest that aucubin facilitates neurite elongation in cultured hippocampal neurons and axonal regrowth in regenerating sciatic nerves. Here, we investigated whether aucubin facilitates the differentiation of neural precursor cells (NPCs) into specific types of neurons. In NPCs cultured primarily from the rat embryonic hippocampus, aucubin significantly elevated the number of GAD65/67 immunoreactive cells and the expression of GAD65/67 proteins was upregulated dramatically by more than three-fold at relatively low concentrations of aucubin (0.01 µM to 10 µM). The expression of both NeuN and vGluT1 of NPCs, the markers for neurons and glutamatergic cells, respectively, and the number of vGluT1 immunoreactive cells also increased with higher concentrations of aucubin (1 µM and 10 µM), but the ratio of the increases was largely lower than GAD expression and GAD immunoreactive cells. The GABAergic differentiation of pax6-expressing late NPCs into GABA-producing cells was further supported in cortical NPCs primarily cultured from transgenic mouse brains, which express recombinant GFP under the control of pax6 promoter. The results suggest that aucubin can be developed as a therapeutic candidate for neurodegenerative disorders caused by the loss of inhibitory GABAergic neurons.

Effects of Different Drying Methods on Contents of 4 Active Ingredients in Leaves of Eucommia ulmoides / 中国药房

OBJECTIVE:To investigate the effects of different drying methods on contents of active ingredients in leaves of Eu-commia ulmoides,and provide reference for establishing its drying methods after habitat harvesting. METHODS:Different drying methods [natural drying in the shade for 72 h,natural drying in the sunlight for 36 h,drying beside or over a fire(60℃6 h,80℃2 h,100℃1 h,120 ℃ 0.5 h),microwave vacuum freeze drying for 12 h,vacuum freeze drying for 12 h] were used for process-ing. HPLC was conducted to determine the contents of aucubin,geniposidic acid,chlorogenic acid and geniposide in samples and compare with the untreated fresh products. RESULTS:Contents of 4 ingredients in samples after the 2 freeze drying were close to these in fresh samples,which were higher than samples after other drying. CONCLUSIONS:Drying methods show significant ef-fects on the effective ingredients in leaves of E. ulmoides. Compared with natural drying in the shade and natural drying sunlight and drying beside or over a fire,microwave vacuum freeze drying and vacuum freeze drying can make better retention of the active ingredients in fresh leaves of E. ulmoides.

Dynamic changes of development and active ingredients content of male flower of Eucommia ulmoides / 中草药

Objective: To establish a method for the determination of six active ingredients in Eucommia ulmoides male flowers, and analyze the dynamic changes of male flower development and active ingredients content of E. ulmoides. Methods: The active ingredients were extracted by the ultrasound-assisted process, the content of total flavonoids was determined by AlCl3 colorimetric method and other six ingredients were determined by HPLC, the samples were separated on a Thermal hypersil gold column (250 mm × 4.6 mm, 5 μm) with gradient elution of methanol-0.5% phosphoric acid at a flow rate of 1.0 mL/min, the UV wavelength was set at 206 nm (0-15 min), 236 nm (15-55 min), and 255 nm (55-100 min). Results: Flower diameter, flower height, fresh weight, dry weight, stamen length and stamen number all reached the maximum at full bloom stage; The contents of total flavonoids, chlorogenic acid and total active ingredients were the highest at the bud stage and the lowest at the initial flowering stage; The content of aucubin showed a tendency of graudal decreasement; The content of geniposidic acid was the lowest at the bud stage and reached the highest at full bloom stage, and decreased at the end of flowering phase; The content of geniposide, isoquercitrin and astragaline were the highest at full bloom stage and the lowest at the initial flowering stage. Conclusion: Flowering stage had a great impact on morphology, yield, active ingredients content and quality of E.ulmoides male flowers, which could provide important information for quality control and product development of E.ulmoides male flowers.

Research progress on chemical structures and pharmacological effects of natural aucubin and its derivatives / 中草药

Aucubin, one of natural active constituents, is widely distributed in Chinese materia medica, such as Scrophularia ningpoensis, Eucommia ulmoides, Plantago depressa and so on, and its pharmacolagical activities have been paid more and more attention. Nevertheless, aucubin derivatives are rarely reported. In this review, the chemical constituents of aucubin derivatives mainly included 6-O-substituent type, 10-O-substituent type, 6,10-O-disubstituent type, 6′-O-substituent type and other types, meanwhile, their pharmacological activities, such as anti-oxidant, anti-inflammatory, antibacterial, neuroprotective and antitumor activities, were summarized in the recent 10 years.

Simultaneous Determination of Aygdalin, Aucubin, Harpagide, Peimisine, Peimine and Peiminine in Keling Capsules by HPLC-ELSD / 中国药师

Objective: To develop an HPLC-ELSD method for the determination of amygdalin, aucubin, harpagide, peimisine, peimine and peiminine in Keling capsules simultaneously.Methods: An Ultimate XB C 18 (250 mm× 4.6 mm , 5 μm) chromatographic column was adopted with an ELSD (the drift tube temperature was 105℃, the flow rate of nitrogen was 2.0 L·min-1).The mobile phase was acetonitrile-methanol (1∶1) and 0.4% acetic acid solution with gradient elution at a flow rate of 0.7 ml·min-1 , and the column temperature was set at 35 ℃.Results: Amygdalin, aucubin, harpagide, peimisine, peimine and peiminine was linear within the range of 13.56-271.20 μg·ml-1 (r=0.999 2), 8.48-169.60 μg·ml-1 (r=0.999 9), 4.89-97.80 μg·ml-1 (r=0.999 7), 2.66-53.20 μg·ml-1 (r=0.999 4), 1.82-36.40 μg·ml-1 (r=0.999 8) and 2.04-40.80 μg·ml-1 (r=0.999 6), respectively.The average recovery and the corresponding RSD were 97.90% (1.20%), 99.21% (1.62%), 97.68% (0.75%), 98.36% (1.38%), 99.70% (0.79%) and 97.95% (1.56%)(n =6), respectively.Conclusion: The method is simple and specific, and the results are accurate and repeatable.The method is helpful to the quality control of Keling capsules.

Effects of Different Drying Methods on Contents of 4 Active Ingredients in Leaves of Eucommia ulmoides / 中国药房

OBJECTIVE:To investigate the effects of different drying methods on contents of active ingredients in leaves of Eu-commia ulmoides,and provide reference for establishing its drying methods after habitat harvesting. METHODS:Different drying methods [natural drying in the shade for 72 h,natural drying in the sunlight for 36 h,drying beside or over a fire(60℃6 h,80℃2 h,100℃1 h,120 ℃ 0.5 h),microwave vacuum freeze drying for 12 h,vacuum freeze drying for 12 h] were used for process-ing. HPLC was conducted to determine the contents of aucubin,geniposidic acid,chlorogenic acid and geniposide in samples and compare with the untreated fresh products. RESULTS:Contents of 4 ingredients in samples after the 2 freeze drying were close to these in fresh samples,which were higher than samples after other drying. CONCLUSIONS:Drying methods show significant ef-fects on the effective ingredients in leaves of E. ulmoides. Compared with natural drying in the shade and natural drying sunlight and drying beside or over a fire,microwave vacuum freeze drying and vacuum freeze drying can make better retention of the active ingredients in fresh leaves of E. ulmoides.

Comparison on contents of four active constituents between slab and branch barks of Eucommia ulmoides / 中草药

Objective: To determine the contents of aucubin, chlorogenic acid, geniposide, and pinoresinol diglucoside in the slab and branch barks of Eucommia ulmoides. Methods: The separation was performed on a Cosmosil C18 (250 mm ×4.6 mm, 5 μm) column with the gradient elution acetonitrile -0.1% phosphoric acid; The flow rate was 0.8 mL/min; The detection wavelength was 208 nm; The column temperature was at 25 ℃. Results: The average content of aucubin: slab bark > branch bark; The average content of chlorogenic acid: branch bark > slab bark; The average content of geniposide: branch bark > slab bark; The average content of pinoresinol diglucosid: slab bark > branch bark. In different origins, the average contents of the above four constituents are more certain differences. Conclusion: The method is simple, rapid, accurate, and there are significant differences in the contents of aucubin, chlorogenic acid, geniposide, and pinoresinol diglucosid from the different parts of E. ulmoides, which would provide the better technique support for the quality control of E. ulmoides.

Aucubin Promotes Neurite Outgrowth in Neural Stem Cells and Axonal Regeneration in Sciatic Nerves

Aucubin is an iridoid glycoside with a wide range of biological activities, including anti-inflammatory, anti-microbial, anti-algesic as well as anti-tumor activities. Recently, it has been shown that aucubin prevents neuronal death in the hippocampal CA1 region in rats with diabetic encephalopathy. In addition, it has protective effects on H2O2-induced apoptosis in PC12 cells. We have shown here that aucubin promotes neuronal differentiation and neurite outgrowth in neural stem cells cultured primarily from the rat embryonic hippocampus. We also investigated whether aucubin facilitates axonal elongation in the injured peripheral nervous system. Aucubin promoted lengthening and thickness of axons and re-myelination at 3 weeks after sciatic nerve injury. These results indicate that administration of aucubin improved nerve regeneration in the rat model of sciatic nerve injury, suggesting that aucubin may be a useful therapeutic compound for the human peripheral nervous system after various nerve injuries.

Protective effects of aucubin on keratinocyte damaged by Ultraviolet B / 中国药学杂志

OBJECTIVE: To investigate the protection of aucubin on keratinocyte damaged by Ultraviolet B and explore its possible mechanism. METHODS: The photo damage model of keratinocyte was established by irradiating of UVB (64 mJ · cm-1). The different concentration aucubin were used for damaged cell. The cell viability was detected by MTT, the SOD activity, GSH-Px activity, CAT activity, MDA content were assayed by kit respectively. The mRNA levels of P38, TNF-α and IL-6 were determined by RT-PCR. The secretion levels of TNF-α and IL-6 of cultured keratinocyte were detected by ELISA. RESULTS: After the UVB irradiation, the cell viability, the activities of SOD, GSH-Px and CAT were decreased, the content of MDA, TNF-α and IL-6, the mRNA levels of P38, TNF-α and IL-6 were increased (P < 0.01). The above changes were inhibited by 1 × 10-1 and 1 × 10-1 mol · L-1 aucubin (P < 0.05 or P < 0.01). CONCLUSION: The aucubin could inhibit the injury by UVB for keratinocyte. And its possible mechanism may have relationship with the inhibition of oxidative damage, regulatory P38 signal pathway, adjusting the expression of TNF-α and IL-6.

Simultaneous determination of five index components in Eucommiae Cortex by HPCE / 中国药学杂志

OBJECTIVE: To establish a high performance capillary electrophoresis (HPCE) method for simultaneous determination of aucubin, geniposide, pinoresinol diglucoside, harpagide, and chlorogenic acid in Eucommiae Cortex. METHODS: 60 mmol·L-1 sodium borate-20 mmol·L-1 sodium dihydrogen phosphate-10% methanol was used as buffer solution (pH 10.0), un-coated fused silica capillary (64.5 cm×75 μm id, 56 cm of effective length) was used, separation voltage was 20 kV, detection wavelength was 210 nm, column temperature was maintained at 25°C, and sample was injected at 5 kPa×6 s. RESULTS: Five index components showed good linearity (r>0.9973) in the range of the tested concentration. The average recoveries of the method were between 96.63%-103.73%. CONCLUSION: The method is simple, accurate and reproducible, and can be used for quality control of Eucommiae Cortex. Copyright 2012 by the Chinese Pharmaceutical Association.

Simultaneous determination of five index components in Rehmanniae Radix by HPCE-DAD / 中国药学杂志

OBJECTIVE: To establish a high performance capillary electrophoresis method with diode array detection (HPCE-DAD) method for simultaneous determination of catalpol, aucubin, geniposide, acteoside and cinnamic acid in raw and processed Rehmanniae Radix. METHODS: 60 mmol · L-1 sodium borate was used as buffer solution (5% methanol, pH 9.5), uncoated fused silica capillary (64.5 cm × 75 μm, 56 cm of effective length) was used, separation voltage was 20 kV, detection wavelength was 210 nm, column temperature was maintained at 25°C, and sample was injected at 5 kPa × 6 s. RESULTS: Five index components showed good linearity (r > 0.9978) in the ranges of the tested concentrations, the average recoveries of the method were between 97.63%-103.03%. CONCLUSION: The method is simple, accurate and reproducible, and can be used for the quality control of Rehmanniae Radix.