Chemical constituents from the fruits of Alpinia oxyphylla and their neuroprotective effects / 药学学报

Fourteen compounds were isolated from the ethyl acetate fraction of 90% EtOH extracts of the dried fruits of Alpinia oxyphylla by silica gel, MCI, RP-18, Sephadex LH-20, TLC and semi-preparative HPLC column chromatography. Their structures were identified by HR-ESI-MS, UV, IR, NMR, ECD and X ray single crystal diffraction spectroscopic data as: (2R,5R,7R,10S)-2,7-dihydroxyl-eudesmane-3(4),11(12)-diene (1), α-rotunol (2), diketone I (3), (1S,4S,5R,7S)-1-hydroxyl-eremophilane-9(10),11(12)-diene-8-one (4), cyperusol A1 (5), (6R,9S,10S)-10-hydroxyl-11,12,13-trinor-cadinane-4(5)-ene-3-one (6), (2E,4E)-6-hydroxy-2,6-dimethylhepta-2,4-dienal (7), oxyphyllacinol (8), yakuchinone A (9), (5R)-5-hydroxy-1,7-diphenylhept-3-heptanone (10), (5S)-5-hydroxy-7-(4″-hydroxyphenyl)-1-phenylhept-3-heptanone (11), (5S)-5-hydroxy-7-(4″-hydroxyl-3″-methoxyphenyl)-1-phenyl-3-heptanone (12), 7-(4″-hydroxy-3″-methoxyphenyl)-1-phenyl-3,5-heptadione (13), bis-(2-ethylhexyl) terephthalate (14). Compounds 1-6 were sesquiterpenoids in which compound 1 is a new eudesmane sesquiterpenoid and compound 7 was a monoterpenoid. Compounds 8-13 were diarylheptanoids, and compounds 2-6 and 14 were isolated from A.oxyphylla for the first time. The experiments on H2O2 induced SH-SY5Y cells showed that compounds 2, 6, 7, 12 and 13 had neuroprotective effects at low and medium concentrations. In particular, compound 6 showed obvious neuroprotective effect at low, medium and high concentrations whose cell viability was higher than that of the positive control.

Mechanism of volatile oil from Alpinia oxyphylla in treating Alzheimer's disease based on GC-MS and network pharmacology / 中国中药杂志

To study the material basis and mechanism of volatile oil from Alpinia oxyphylla in treating Alzheimer's disease(AD) based on GC-MS and network pharmacology. Ingredients of volatile oil from A.oxyphylla were analyzed by GC-MS. Targets of those ingredients were obtained through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). Relevant targets of AD were obtained through such databases as DrugBank, STITCH, OMIM. Intersection targets of ingredients and diseases were obtained by Online Venny map, and PPI network was established by STRING to screen out core targets. Gene ontology(GO) functional enrichment analysis and KEGG pathway enrichment analysis were performed by DAVID. The "ingredients-target-pathway" network was constructed by software Cytoscape 3.8.1 to screen out potential active ingredients of volatile oil from A.oxyphylla in the treatment of AD. The results showed that a total of 6 active ingredients were screened from the volatile oil of A.oxyphylla by GC-MS, 17 targets corresponding to 6 active ingredients were found in TCMSP database, and 3 448 AD targets were found in DrugBank database. "Ingredients-target-pathway" network and PPI network showed there were 4 potential active ingredients in the treatment of AD and 4 core targets. GO analysis and KEGG analysis showed 34(P<0.05) and 5(P<0.05) pathways, respectively, including nerve ligand receptor interaction, calcium signaling pathway, cholinergic synapse and 5-hydroxytryptaminergic synapse. This suggested that volatile oil from A.oxyphylla could synergistically treat AD by regulating calcium balance, cholinergic balance and phosphorylation. This study provided reference and guidance for further study of volatile oil from A.oxyphylla in the treatment of AD.

Chemical constituents from Alpinia oxyphylla / 中草药

Objective: To study the chemical constituents of Alpinia oxyphylla. Methods: The ethyl acetate fraction of 95% ethanol extract from A. oxphylla was isolated and purified by silica, MCI, Sephadex LH-20 and semi-preparative HPLC, then the structures of obtained compounds were identified by physicochemical properties and spectral data. Results: Sixteen compounds were isolated and their chemical structures were identified as phthalic acid-bis (2’-ethyl heptanyl) easter (1), (E)-1-(4’-hydroxy-3’-methoxyphenyl)-7- (4″-hydroxy-phenyl)-hept-4-en-3-one (2), 5-hydroxy-7-(4″-hydroxy-3″-methoxyphenyl)-1-phenyl-3-heptanone (3), 1β,4β,7β- trihydroxyeudesmane (4), bullatantriol (5), 1,5-epoxy-3-hydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-7-(4-hydroxy-3-methoxy- phenyl) heptanes (6), 1-tetratriacontanol (7), dihydrogingerenone B (8), tectochrysin (9), chrysin (10), oxyphyllenone B (11), (1R,4R,10R)-1β,4α-dihydroxy-11,12,13-trinor-5,6-eudesmen-7-one (12), daucosterol (13), 1-(4’-hydroxyphenyl)-7-(3″-methoxy- 4″-hydroxyphenyl)-4-ene-3-heptanone (14), yakuchinone A (15) and 5-dehydroxy-hexahydro-demethoxycurcumin B (16). Conclusion: A total of 16 compounds were isolated and identified. The compounds 1-2, 4-8 are isolated from the genus for the first time, and compounds 3 were isolated from the plant for the first time.

Analysis and Identification of Sesquiterpenes in Alpinia oxzphylla Miq Based on UPLC-Q-Orbitrap HRMS / 中草药

Objective: To establish a rapid and accurate analytical method for the identification of complex system of traditional Chinese medicine, and to systematically clarify the chemical composition of sesquiterpenes in Alpinia oxyphylla. Method: On the basis of optimizing the extraction process of sesquiterpenes, the accurate molecular weight and secondary fragment ions information of unknown compounds were captured by ultra-high performance liquid chromatography-quadrupole/orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS). Compared with the relative retention time and mass spectrometry data of the reference substance, combined with relevant references and databases, the sesquiterpene unknown compounds in the fruits of A. oxyphylla were accurately and rapidly characterized. Results: A total of 24 sesquiterpenes were identified and classified into four categories according to their skeleton structure, including nine eudesmane-type, six cadinane-type, eight eremophilane-type, and one oplopanone-type. Conclusion: In this study, the established analytical method was used to realize the rapid and accurate identification of sesquiterpenes in the fruits of A. oxyphylla, which provided a theoretical basis for the research on the pharmacodynamic substance basis and quality control of the fruits of A. oxyphylla.

Protective effect of tropical herb-Alpinia oxyphylla compound formula on diabetic nephropathy mice

Objective: To observe the mechanism of Alpinia (A.) oxyphylla in the treatment of diabetic nephropathy by regulating blood glucose level. Methods: A total of 40 db/db diabetic nephropathy mice were randomly divided into four groups including model group, irbesartan group, A. oxyphylla compound formula high dose group and A. oxyphylla compound formula low dose group with 10 in each group. Ten normal mice served as normal control group. Normal group and model group were given normal saline only. Irbesartan group and A. oxyphylla compound formula high and low dose group were given corresponding drugs once a day for 4 weeks (about 0.2 mL, once a day). Blood glucose, BUN, urine protein, SCr excretion, GSH and CAT activity were detected. Results: Compared with normal group, the blood glucose of model group were increased significantly (P<0.05). While compared with model group, the blood glucose of irbesartan group and A. oxyphylla compound formula groups (both high and low dose) decreased significantly after 4 weeks' treatment. Compared with model group, 24 h urine protein, BUN and SCr of A. oxyphylla compound formula groups (both high and low dose) decreased significantly(P<0.05) after 4 weeks' treatment. Compared with normal group, GSH, CAT of model group decreased significantly (P<0.05). While compared with model group, GSH, CAT of irbesartan group and A. oxyphylla compound formula high dose group increased significantly (P<0.05). Conclusions: A. oxyphylla compound formula can protect diabetic nephropathy mice by reducing the blood glucose level, decreasing the excretion of urine protein, BUN, SCr, and increasing the activites of CAT and GSH.

Alpinia oxyphylla alleviates diabetic nephropathy through the NLRP3 inflammasome pathway in rats

Objective: Previous studies have confirmed that the tropical traditional Chinese herb-Alpinia (A.) oxyphylla are effective on diabetic nephropathy. The purpose of this experiment was to investigate whether A. oxyphylla could prevent and treat diabetic nephropathy by regulating NLRP3 inflammasome pathway. Methods: Male SD rats and NLRP3 gene silencing rats were assinged into blank group, model group, A. oxyphylla group, valsartan group and NLRP3 gene silencing group, with 8 rats in each group. The model was established by intraabdominal injection of streptomycin after feeding with high sugar and high fat diet for 6 weeks. Based on successful modeling, administration protocal for a period of 8 weeks were performed. A. oxyphylla group was given A. oxyphylla decoction (2.6g/kg/d ), while the model group and NLRP3 gene silencing group were given saline as the same dosage of A. oxyphylla decoction. The valsartan (10 mg/kg/d) was given to the valsartan group. After 8 weeks of treatments, blood glucose, 24 h urinary protein, creatinine, urea nitrogen levels were detected, and the pathological sections of the kidney were examined. The expression of NLRP3 protein was detected by Western-Blot and PCR. NLRP3 protein, caspase-1, IL-18 and IL-1βin renal tubules were detected in each group by immunofluorescence and immunohistochemistry. Results: Compared with the model group, A. oxyphylla significantly reduced blood glucose, 24 h urinary protein, creatinine, urea nitrogen levels (P all<0.05) but the pathological injury in kidney was less. The expression of NLRP3 protein was negative in NLRP3-/-group while positive in the model group, A. oxyphylla group and valsartan group. Expression of NLRP3 protein, NLRP3, caspase-1, IL-18 and IL-1β were also significantly lower in A. oxyphylla trated group compared with the model group (P all<0.05). Conclusions: A. oxyphylla could inhibit the expression of NLRP3 protein, reduce IL-18 and IL-1β and other inflammatory factors and alleviate the damage of kidney caused by inflammatory reaction. This is one of the mechanisms whereby A. oxyphylla prevents and treats diabetic nephropathy.

Present situation and development strategy of Alpinia oxyphylla / 中国中药杂志

@#Alpinia oxyphylla is mainly produced in Hainan,and also one of the four famous traditional Chinese medicines in South China with increasing importance in traditional Chinese medicine industry. Field surveys and literatures show that A. oxyphylla has widely used as a medicinal and edible plant,it is an important raw material for many Chinese patent medicines,health products and food,with a long history of artificial cultivation and application. The future development is prospected on health market. But A. oxyphylla industry has faced a lot of problems,including unreasonable planting layout,lack of good varieties,imperfect seed breeding system,low level of standardization,inconsistent quality of medicinal materials,low level of industry,and so on. The suggestions for sustainable development are listed below.First,it is essential to strengthen the research on the basis and application technology of A. oxyphylla,speed up the selection and breeding of improved varieties,and popularize standardized cultivation techniques. Secondly,it is important to strengthen the research on quality standards,improve the quality evaluation system of medicinal materials. Thirdly,it is necessary to take full advantage of the functional components to develop functional products with Hainan characteristics,find out the unique product characteristics of A. oxyphylla,build a famous brand and improve the product competitiveness in the market. It is also important to strengthen policy support and industrial supervision,promote the healthy and rapid development of A. oxyphylla industry.

Alpinia oxyphylla Miq. extract changes miRNA expression profiles in db-/db- mouse kidney

BACKGROUND: A number of dysregulated miRNAs have been identified and are proposed to have significant roles in the pathogenesis of type 2 diabetes mellitus or renal pathology. Alpinia oxyphylla has shown significant anti-inflammatory properties and play an anti-diabetes role. The objective of this study was to detect the alteration of miRNAs underlying the anti-diabetes effects of A. oxyphylla extract (AOE) in a type II diabetic animal model (C57BIKsj db-/db-). RESULTS: Treatment with AOE for 8 weeks led to lower concentrations of blood glucose, urine albumin, and urine creatinine. 17 and 13 miRNAs were statistically identified as differentially regulated in the DB/DB and db-/db- AOE mice, respectively, compared to the untreated db-/db- mice. Of these, 7 miRNAs were identified in both comparison groups, and these 7 miRNAs were verified by quantitative real-time PCR. Functional bioinformatics showed that the putative target genes of 7 miRNAs were associated with several diabetes effects and signaling pathways. CONCLUSIONS: These founding suggest that the potential of AOE as a medicinal anti-diabetes treatment through changes in the expressions of specific miRNAs. The results provide a useful resource for future investigation of the role of AOE-regulated miRNAs in diabetes mellitus.

Effects of Extracts from Different Polar Parts of Li Medicine Alpinia oxyphylla Fruit on Model Mice with Experimental Colitis / 中国药房

OBJECTIVE:To investigate the effects of 4 polar parts extracts from Li medicine Alpinia oxyphylla fruit on model mice with experimental colitis,and screen the effective parts of A. oxyphylla fruit in the treatment of colitis. METHODS:After 70% ethanol extract dispersed by water,petroleum ether,trichloromethane,ethyl acetate and n-butanol were used in turn to obtain extractions in related parts. 42 mice were randomly divided into modeling group(36 mice)and blank group(6 mice). Mice in mod-eling group were taken 2,4,6-nitrobenzne sulfonic acid method to replicate the experiment colitis. After modeling,model mice were randomly divided into model group,positive group(Sulfasalazine enteric coated tablets,52 g/kg),petroleum ether,trichloro-methane,ethyl acetate,n-butanol extract from A. oxyphylla fruit groups(10 g/kg,calculated by crude drug),6 in each group,in-tragastrically administrated once a day,for 9 d,0.2 mL/10 g;mice in blank group and model group were intragastrically given nor-mal saline. After administration,body mass of mice was determined,disease activity index(DAI)was scored and colonic myelo-peroxidase(MPO),superoxide dismutase(SOD),malondialdehyde(MDA)activities in colon tissue were detected,and the colon pathological changes were observed and scored. RESULTS:Compared with blank group,decrease percentage of body mass,DAI score,MPO and MDA activities in colon tissue in model group were increased(P<0.05 or P<0.01);colon tissue was pathologi-cally changed. Compared with model group,body mass was increased,DAI score,MPO activity and pathological score in each ad-ministration groups were decreased;SOD activities in positive group,n-butanol extract from A. oxyphylla fruit group and ethyl ace-tate extract from A. oxyphylla fruit group were significantly increased,MDA activities in positive group and ethyl acetate extract from A. oxyphylla fruit group were significantly decreased,with statistical significances (P<0.05 or P<0.01). CONCLUSIONS:The 4 polar parts extracs from A. oxyphylla fruit have certain improvement effects on model mice with experimental colitis, in which ethyl acetate and n-butanol parts are preferred.

Examination of CNS Activity of Rhizomes of Alpinia Oxyphylla.

The study is based on the examination of the CNS activity observed from the methanolic extract of the rhizomes of Alpinia oxyphylla. Tail immersion method in mice has been used for the evaluation of the central pharmacological actions. Similarly acetic-acid induced writhing-test was used for the evaluation of the peripheral pharmacological properties. A significant rise in pain threshold is seen in a dose dependent manner with the methanolic extract of A. oxyphylla at doses of 100, 200 and 400 mg/kg body weight with the tail immersion methods. The methanolic extract at 400 mg/kg dose possessed 73.12% writhing inhibition, (p <0.001) in acetic-acid induced writhing-test that could be compared to the standard, Diclofenac-Na (25 mg/kg) with 75.78% inhibition. Open-field and hole-cross tests have been conducted in mice for further investigation of the extract in support of its neuro-pharmacological actions, where dosedependent suppression of exploratory and motor activities were observed in the tested models. Hence, the above results evidence the presence of CNS depressant and analgesic properties of the plant, A. oxyphylla.

Simultaneous quantitation of four antioxidant components in Alpinia oxyphylla Miq. by UPLC / 中国药学杂志

OBJECTIVE: To establish a UPLC method for simultaneous determination of 5-hydroxymethyl furfural (1), leu-hetenone A (2), lectochrysin (3), and nootkatone (4) in Alpinia, oxyphylla Miq, and to compare the contents of the four components in different parts of this medicinal materials from different places. METHODS: The UPLC method was established on an HSS T3 Column (2.1 mm × 100 mm, 1.8 μm). The mobile phase consisted of water containing 0.1% formic acid and acetonilrile in gradient elution mode at the flow rate of 0.5 mL · min-1 and the detection wavelength was set at 255 nm. RESULTS: The standard curves of compounds 1, 2, 3, and 4 showed good linearity in the ranges of 1.223-24.46, 2.016-40.32, 1.875-37.50 and 16.78-335.6 μg · mL-1 with the corresponding average recoveries of 99.5%, 101.5%, 100.9% and 101.2%, respectively. CONCLUSION: The method is precise and highly reproducible, which can be used to simultaneously determine the antioxidant components including 5-hydroxymelhyl furfural, teuhetenone A, tectochrysin, and nootkatone in Alpinia oxyphylla Miq; compounds 2 and 4 are mainly extracted from the seeds, while compounds 1 and 3 come from the seeds and putamens equally. There are significant differences among the samples from different production areas.

Effects of Alpinia oxyphylla fructus on learning-memory and expression of related signal proteins in hippocampus of brain aging mice / 中华行为医学与脑科学杂志

Objective To investigate the effects and mechanisms of Alpinia oxyphylla fructus (AOF) on learning and memory in D-galactose induced brain aging mice. Methods The brain aging model was induced by s. c D-galactose. Learning-memory ability was tested by passive avoidance test and Morris water maze test, and the expression of synapsin ( Syn), mitogen-activated protein kinase (MAPK) and protein kinase ( PKC ) in hippocampus were examined by Western blot. Results ① Passive avoidance test:the latency in brain aging group( ( 119.80 ±101.80)s) significantly decreased,and the number of errors (4.4 ± 1.3 ) significantly increased compared with the control group( latency: (279.30 ± 31.64) s; number of errors: ( 1. 8 ±0.9), P＜0. 01 ) ). The latency in low dose, middle dose and high dose AOF group( ( 170.25 ± 68.31 ) s, (226.31 ± 73.25 ) s, (263.20 ± 70.55 ) s) significantly increased, and the number of errors in middle dose and high dose AOF group ( ( 2.8 ± 1.2 ), ( 2.3 ±0. 9 ) ) significantly decreased compared with brain aging group (P ＜ 0. 05, P ＜ 0. 0 1 ). ② Morris water maze test:the escape latency in brain aging group was significantly longer, and the time spent in the original quadrant that previously contained the platform was significantly shorter compared with the control group (P＜0. 01 ). The escape latency in 3 AOF groups was significantly shorter (P＜ 0. 05 ), and the time spent in the original quadrant that previously contained the platform in middle and high dose AOF groups was significantly longer compared with brain aging group (P＜0. 05, P＜0. 01 ). ③ Western blot test:the expression of Syn,MAPK and PKC in hippocampus of brain aging group was significantly weakened than that of the control group. In contrast, the expression of Syn,MAPK, PKC were significantly enhanced in all AOF groups. Conclusion AOF could significantly improve the ability of learning and memory in brain aging mice. Its effects might be related to the increase of the expression of Syn, MAPK and PKC in hippocampus.

Effect of water extracts of Alpinia oxyphylla Miq. fruit on memory impairment in cerebral ischemia rats / 中华行为医学与脑科学杂志

Objective This report investigate the intervention effect of the water extract of Alpinia oxyphylla Miq. fruit (AOF)on memory impairment and the mechanism in cerebral ischemia rats. Methods 48 Rats were randomly divided into sham-operated group(n=12), ischemia group(n=12), AOF group Ⅰ( n= 12)and AOF group Ⅱ (n= 12). The model of transient cerebral ischemic/reperfusion was made by bilateral occlusion of common carotid arteries in rats and combination with reducing blood pressure with an abdominal injection of so-dium nitroprusside. Learning-memory ability was observed by step through test. The content of NO and the activi-ties of NOS were measured in hippocampus. Results In step through test,the latency in ischemia group[(143.8±65.2)s]significantly decreased, the number of errors (8.9±4.2 ) significantly increased compared with sham-operated group [latency: (257.2±67.1 ) s; number of errors: (1.7±1.1 ), P<0.01 ]. The latency in AOF group Ⅰ and AOF group Ⅱ[(186.5±46.2) s, (193.4±43.7 ) s ] significantly increased, the number of errors (6.1±2.9,5.2±2.1 ) significantly decreased compared with ischemia group(P<0.05, P<0.01). In hippocampus, the content of NO and the activities of NOS in ischemia group [(56.53±27.42) nmol/mg prot, (17.23±5.64) nmol/mg prot] significantly increased compared with sham-operated group[ (40.02±17.9 ) nmol/mg prot, ( 10.46±6.15)nmol/mg prot], and in AOF group Ⅰ and AOF group Ⅱ [content of NO:group Ⅰ (46.60 ±20. 26)nmol/mg prot,group Ⅱ (42.38±21.23) nmol/mg prot ;activities of NOS:group Ⅰ (13.98±5.13 ) nmol/mg pint,group Ⅱ(13.61±5.27) nmol/mg prot] significantly decreased compared with ischemia group(P<0.05). Conclu-sion AOF could significantly ameliorate the memory impairment in cerebral ischemic rats. Its effects may be in-volved in the decrease of content of NO and activities of NOS in hippocampus.