Acanthopanax senticosus Protects Structure and Function of Mesencephalic Mitochondria in A Mouse Model of Parkinson's Disease / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the neuro-protective effects of Acanthopanax senticosus Harms (EAS) on mesencephalic mitochondria and the mechanism of action, using a mouse model of Parkinson's disease (PD).</p><p><b>METHODS</b>The chemical fingerprint analysis of the extract of Acanthopanax senticosus Harms (EAS) was performed using the ultra performance liquid chromatograph and time of flight mass spectrometry. Thirty mice were randomly divided into the control group, the MPTP model group, and the EAS treated group with MPTP (MPTP+EAS group, 10 in each group). The MPTP model group and the MPTP+EAS group received MPTP-HCl (30 mg/kg i.p) once a day for 5 days. The control group received an equal volume of saline (20 mL/kg i.p) once a day for 5 days. Induced by 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine hydrochloride daily (MPTP-HCl, 30 mg/kg) for 5 days, the PD mice were treated with EAS at 45.5 mg/kg daily for 20 days. The behavioral testing of mice was carried out using the pole-climbing test. The integrity and functions of neurons were examined in mesencephalic mitochondria in a PD mouse model, including nicotinamide adenine dinucleotide dehydrogenase ubiquinone flavoprotein 2 (NDUFV2), mitochondrially encoded nicotinamide adenine dinucleotide dehydrogenase 1 (MT-ND1), succinate dehydrogenase complex subunit A (SDHA), and succinate dehydrogenase cytochrome b560 subunit (SDHC).</p><p><b>RESULTS</b>After treatment with EAS, the behavioral changes induced by MPTP were attenuated significantly (P<0.05). EAS protected the mesencephalic mitochondria from swelling and attenuated the decreases in their membrane potential (both P<0.05), which was supported by an ultra-structural level analysis. The changes in reactive oxygen species (ROS), malonic dialdehyde (MDA), oxidative phosphorylation (OXPHOS) system 4 subunits levels and PD-related proteins expressions (parkin, Pink1, DJ-1, α-synuclein, and Lrrk2) reverted to near normal levels (all P<0.05), based on the results of immune-histological and Western blotting observations.</p><p><b>CONCLUSIONS</b>The neuro-protective effects of EAS are linked to protecting mice against MPTP-induced mitochondrial dysfunction and structural damage. Therefore, EAS is a promising candidate for the prevention or treatment of mitochondrial neurodegenerative disorders, such as PD.</p>

Inhibitory activity of kaurane-type diterpenes isolated from Acanthopanax senticosus Harms on protein tyrosine phosphatase 1B / 中国药学杂志

OBJECTIVE: To study the constituents isolated from Acanthopanax senticosus Harms and their inhibitory activity on protein tyrosine phosphatase 1B(PTP1B). METHODS: Kaurane-type diterpenes with PTP1 B inhibitory activity were obtained by bio-assay-guided fractionation. RESULTS Nine compounds were identified as 17-isobutyryloxy-16αH-kauran-19-oic acid(1), 17-hydrox-y-16αH-kauran-19-oic acid(2), 17-acetoxy-18-isobutyryloxy-16αH-kauran-19-oic acid (3), ent-kaur-16-en-19-oic acid (4), ent-kaur-16-en-19-oic acid (kaurenoic acid 5), 4-epirulopezol(6), 16α-hydroxy-ent-kauran-19-oic acid(7), 16αH, 17-isovaleryloxy-ent-kauran-19-oic acid(8) and 16α-hydroxy-17-isovaleryloxyent-kauran-19-oic acid(9). CONCLUSION: Compounds 1-3, 5 and 6 are obtained from the plant for the first time. Compounds 1,3-6 and 8 exhibite inhibitory effects on PTP1 B with IC50 values ranging from (5.6 ± 0.8) to (22.8 ± 1.2) μmol · L-1.

Food and Drug Interactions: Effect of <i>Acanthopanax senticosus</i> Harms on CYP2C9 Activity (Part 2) / 日本補完代替医療学会誌

<b>Objective:</b> <i>Acanthopanax senticosus</i> Harms extract (ASE) is an ingredient of functional foods, such as health supplements, in Japan. We investigated the effects of ASE on CYP2C9 activity.<br> <b>Methods and Results:</b> CYP2C9-catalyzed diclofenac 4′-hydroxylase activities in human intestinal and liver microsomes (abbreviated as HIM and HLM, respectively) were significantly decreased by the addition of ASE in a concentration-dependent manner. Kinetic studies of diclofenac 4′-hydroxylase in HLM revealed that ASE addition significantly decreased <i>V</i>_max but had no effect on <i>K</i>_m. These results suggest that diclofenac 4′-hydroxylase activity is suppressed by ASE addition in a non-competitive manner. Then, we investigated the time courses of diclofenac 4′-hydroxylase activity in rat liver microsomes after ASE oral administration (50 to 400 mg/kg). Diclofenac 4′-hydroxylase activities were significantly lowered by the administration of 200 and 400 mg/kg ASE at 0.5 to 4 hr compared with control (0 hr). Furthermore, we investigated the effects of ASE oral administration on the pharmacokinetics of tolbutamide (substrate for CYP2C9) in rats. The area under the concentration-time curve of tolbutamide after ASE oral administration (400 mg/kg) was enhanced by approximately 1.6 times compared with that without ASE oral administration.<br> <b>Conclusion:</b> These findings indicated that ASE inhibits human intestinal and hepatic CYP2C9 activities.<br>

Food and Drug Interactions: Effect of <i>Acanthopanax senticosus</i> Harms on CYP3A4 and CYP2C9 Activities (Part 3) / 日本補完代替医療学会誌

<b>Objective:</b> By using human liver microsomes (HLM), we analyzed the effects of 14 known components of <i>A.senticosus</i> Harms on the activities of CYP2C9 and CYP3A4.<br> <b>Methods and Results:</b> Sesamin and quercetin inhibited both enzyme activities, whereas quercitrin strongly inhibited CYP3A4 activity. The 50% inhibitory concentrations (IC₅₀s) of sesamin and quercetin on CYP2C9 activity were approximately 124- and 59-fold higher and the IC₅₀s of sesamin, quercetin, and quercitrin on CYP3A4 activity were approximately 427-, 135-, and 22-fold higher than that of <i>A. senticosus</i> Harms extract (ASE), respectively. All these components inhibited both CYP3A4 and CYP2C9 in a non-competitive manner. However, these components are present in small amounts in ASE.<br> <b>Conclusion:</b> Therefore, the food-drug interactions caused by <i>A. senticosus </i>Harms are presumed to be due to the additive or synergistic interaction of these components or the other existing components, including their metabolites.<br>

Pharmacodynamic Optimization of Purification Technique for Acanthopanax Senticosus Harms / 中国药房

OBJECTIVE: To optimize the purification technique for the extract of Acanthopanax senticosus Harms. METHODS: The techniques of D- 101 macroporous resin adsorption, polyamide adsorption and n- butanol extraction were separately applied to purify the aqueous extract of Acanthopanax senticosus Harms. Syringin was detected by HPLC and total flavone was detected by UV. The anti- cerebral ischemia effects of three purified samples were compared using rat MCAO model and mouse decapitation gape model. RESULTS: The three purified samples all showed anti- cerebral ischemia effects, with the sample purified via D- 101 macroporous resin adsorption having stronger effects than the other two. The content of Syringin was correlated with anti- cerebral ischemia effects. CONCLUSIONS: D- 101 macroporous resin adsorption is a be-tter technique for purifying the aqueous extract of Acanthopanax senticosus Harms.

Somatic cell embryogenesis of Acanthopanax senticosus / 中草药

Objective To study somatic cell embryogenesis of Acanthopanax senticosus induced by different concentration of 2,4D and type of explants,which provides theoretic evidence in protection of A.senticosus resources and genetic engineering.Methods Using 3-week seedlings and zygotic embryos(cotyledon, hypocotyls,and roots) of stratificated seeds as explants researches the effect of different hormones on somatic cell embryogenesis of A.senticosus. Results Explants of zygotic embryos of stratificated seeds cultured on MS and 1/2MS media containing 0.5 mg/L 2,4-D generated the highest frequency(57.1%) and the most number(3.3) of somatic cell embryos,which can develop into maturation in the initial medium.But it is more beneficial to generate new somatic cell embryos and to develop primary somatic cell embryos into maturation when transferred into 2,4-D of decreased concentration.And the deve-(lopment) process of somatic cell embryos of A.senticosus is similar to that of zygotic embryos.Conclusion(Somatic) embryogenesis of A.senticosus is realized by culturing explants of zygotic embryos and the inductive rate of somatic cell embryos is related to the concentration of 2,4-D and developmental stage of explants.

Comparison of total flavone between root-extract and stem-extract from Acanthopanax / 中成药

AIM:To understand the differences between the extraction parameters of different parts from Acanthopanax. METHODS: The orthogonal design method was used in experiment arrangement; the influence caused by the different factors on the extraction of total flavone from the root and the stem of Acanthopanax senticosus Harms was obtained. RESULTS: The optimum extracting technical parameters for the root were as follows: 20 times of 50% ethanol to Acanthopanax senticosus Harms, refluxing for 3 h at 60 ?C . The optimum extracting technical parameters for the stem were as follows: 20 times of 50% ethanol to Acanthopanax senticosus Harms, refluxing for 4 h at 70 ?C. CONCLUSION: The optimum extracting technical parameters for the root were different from the stem, which is related to difference in the biological structures between the root and the stem of Acanthopanax senticosus Harms.

Purification of Acanthopanax root extract with anticerebral ischemia / 中成药

AIM:To optimize the purification of Acanthopanax senticosus Harms root extract based on the rodent with anticerebral ischemia. METHODS:We identified the effect of D-101 macroporous resin adsorption,polyamide adsorption and n-butanol extraction on cerebral ischemia-reperfusion injure model rat and behead-mouth(opening) model mouse in the treatment of aqueous acanthopanax root extract. RESULTS:Aqueous acanthopanax root extract,which was purfied by using D-101 macroporous resin,had advantage over that were treated with polyamide adsorption and n-butanol extraction.Eleutheroside content in the root extract showed the positive correlation with anticerebral ischemia. CONCLUSION:D-101 macroporous resin adsorption is a good purification method for acanthopanax root extract.