Study on mechanism of hydroxy-a-sanshool on diabetic cardiomyopathy based on proteomics / 中国药理学通报

Aim To explore the mechanism of hydroxy-a-sanshool in the treatment of diabetic cardiomyopathy ( DCM) based on label-free quantitative proteomics detection technique. Methods DCM model was established by high fat diet and intraperitoneal injection of streptozotocin ( STZ) . They were divided into control group ( CON group ) , diabetic cardiomyopathy group (DCM group) and hydroxy-a-sanshool treatment group ( DCM + SAN group) . The cardiac function of mice was evaluated by echocardiography, the myocardial morphology was observed by pathology staining, the protective mechanism of hydroxy-a-sanshool on diabetic cardiomyopathy was speculated by proteomic technique , and the expression level of cAMP/PKA signaling pathway and key proteins were verified by Western blotting. Results Cardiac ultrasound and pathology staining showed that hydroxy-a-sanshool had protective effect on the heart of DCM mice. Label-free quantitative proteomic analysis was carried out between DCM + SAN group and DCM group, and 160 differential pro-teins were identified by proteomics, in which 127 proteins were up-regulated and 33 proteins were down regulated ; GO secondary functional annotations showed the biological process, molecular function and cellular component; KEGG enrichment analysis showed that cAMP signaling pathway was the most abundant; protein interaction network showed that PKA as the central node interacted with many proteins in the cAMP signaling pathway. Western blot showed that the relative expression of с AMP, PKA protein in DCM group was significantly lower than that in CON group ( P < 0. 05 ) , while the relative expression of cAMP, PKA protein in DCM + SAN group was significantly higher than that in DCM group ( P < 0. 05 ) . Conclusions Hydroxy-a-sanshool has protective effect on heart function of mice with diabetes, which plays a role through cAMP signaling pathway.

Zanthoxylum bungeanum Aqueous Extract Treated Rheumatoid Arthritis by Inhibiting Synovial Cell Proliferation and Inducing Cell Apoptosis / 中国实验方剂学杂志

ObjectiveTo investigate the therapeutic effect of the water extract of Zanthoxylum bungeanum aqueous extract（ZBAE）on rheumatoid arthritis. MethodThe sixty SD rats were divided into normal group， model group ［complete Freund's adjuvant （CFA）， 10 mg·kg-1］， methotrexate（MTX） group （0.25 mg·kg-1）， low -， medium -， and high-dose ZBAE groups （90， 180， 360 mg·kg-1）. The rats in MTX group were given intraperitoneal injection for two weeks， three times a week， and the rats in ZBAE group were administrated for 14 days. The swelling of the ankle joint and body weight were observed， and arthritis scores were also performed. Computed tomography （CT）， hematoxylin-eosin （HE） staining and safranine-O and fast green staining were used to observe the effect of ZBAE on synovial hyperplasia and bone protection. Cell counting kit-8（CCK-8）method was used to detect the proliferation of the RA-FLSs cells treated with ZBAE. According to the results of CCK-8 experiment， the optimal concentration and time of administration were determined， blank group， low -， medium -， and high-dose ZBAE groups （0.08，0.10，0.12 g·L-1） were set up. The cell cycle distribution and apoptosis rate were analyzed by flow cytometry，the migration ability of RA-FLSs cells was examined by scratch test. Western blot was used to detect the effect of ZBAE on phosphatidylinositol 3-kinase （PI3K）， protein kinase B （Akt）， cyclin-dependent kinase 2 （CDK2）， Cyclin A and phosphorylated PI3K， Akt （p-PI3K，p-Akt） protein expression in RA-FLSs cells. ResultCompared with the normal group，joint swelling index and arthritis score were increased in the model group （P<0.05），the bone of the ankle was seriously damaged， and there was obvious synovial hyperplasia. Compared with the model group， the ZBAE group could significantly reduce the joint swelling index （P<0.05）， inhibit synovial hyperplasia and cartilage destruction. In vitro study showed that compared with the blank group， ZBAE could inhibit the migration of RA-FLSs （P<0.05）， promoted cell apoptosis （P<0.05）， and acted on RA-FLSs cells in S phase to inhibit cell proliferation. Moreover， the result of Western blot showed that compared with the blank group， the expression of p-PI3K， p-Akt， CDK2 and Cyclin A proteins were significantly decreased in the high dose group of ZBAE （P<0.05）. ConclusionThese results suggest that ZBAE has a therapeutic effect on rheumatoid arthritis by inhibiting synovial hyperplasia， promoting synovial apoptosis and inhibiting its migration.

Chemical Constituents from Stems of Zanthoxylum bungeanum / 中国实验方剂学杂志

Objective::To isolate and identify the chemical constituents from the 95%, 75%ethanol extracts of the stems of Zanthoxylum bungeanum. Method::The 25 kg stems of Z. bungeanum were extracted with 95%, 75%ethanol for three times, and the combined filtrates were concentrated under vacuum to get the extracts. The 95%extracts were then extracted by petroleum ether, dichloromethane, ethyl acetate and n-butanol successively to obtain corresponding fractions. Such fractions and 75%extracts were isolated and purified by silicagel, Sephadex LH-20, ODS, preparation HPLC and recrystallization to obtain compounds. Their structures were identified by mass spectroscopy (MS), and nuclear magnetic resonance (NMR). Result::Sixteen compounds were isolated from the stems of Z. bungeanum and identified as dictamnine(1), decarine(2), zanthobungeanine(3), pseudocolumbamine(4), skimmianine(5), norchelerythrine(6), osthenol(7), dimethylfraxetin(8), 3, 4, 5-trimethoxycinnamylalcohol(9), asarinin(10), yangambin(11), syringaresinol(12), ashantin(13), bis(2-ethylhexyl) benzene-1, 2-dicarboxylate(14), 24-propylcholesterol(15), and sucrose(16). Conclusion::Compounds pseudocolumbamine(4), 3, 4, 5-trimethoxycinnamylalcohol(9), and 24-propylcholesterol(15)were isolated from the genus of Zanthoxylum for the first time and compounds dictamnine(1), osthenol(7), dimethylfraxetin(8), asarinin(10), yangambin(11), syringaresinol(12), ashantin(13), and bis(2-ethylhexyl) benzene-1, 2-dicarboxylate(14)were isolated from this plant for the first time.

Transcriptome and quality evaluation of Sichuan Dao-di herbs Zanthoxylum bungeanum / 中国中药杂志

"Huajiao" is dried ripe fruit peel of Zanthoxylum bungeanum or Z. schinifolium, is konwn as geoherbs, especially the "Dahongpao" cultivated in Hanyuan, Maoxian and Jiulong of Sichuan province. However, the genetic basis of Dao-di "Huajiao" is virtually unknown. The transcriptome of the fruit and leaf from Sichuan(Hanyuan, Jiulong, Lixian, Maoxian), Gansu(Wudu) province and Shaanxi(Fengxian) province was sequenced. Trinity de novo assembling resulted in a total of 177 616 unigenes. Through the KEGG, NR, SwissProt, Trembl, KOG/COG, GO, Pfam database comparision 106 644 annotated Unigene finally, 4 574 deferentially expressed genes were found in fruit between Sichuan and other provinces, including 3 740 up-regulated genes and 834 down-regulated genes. Among the up-regulated genes, 27 up-regulated genes were raleted to terpenoids, and 8 up-regulated genes were related to isoquinoline alkaloid bio-synthesis. Furthermore, it was also showed remarkable differences in groups which enrichment ratio of the diffe-rent expressed gene compared. The different expressed genes were annotated by the KEGG database into plant-pathogen interaction, plant hormone signal transduction and phenylpropanoid biosynthesis in fruit and leaf, but isoflavonoid bio-synthesis and betaine bio-synthesis were significantly different in fruit and leaf. The study laid a certain reference basis for comparison of quality and different expressed gene of Z. bungeanum from different groups.

Chemical Constituents of Pericarps of Zanthoxylum bungeanum / 中国实验方剂学杂志

<b>Objective::To study the chemical constituents of pericarps of <italic>Zanthoxylum bungeanum</italic>. <b>Method::The dried pericarps of <italic>Z</italic>. <italic>bungeanum</italic> were smashed, and then extracted and concentrated in 95%ethanol to obtain the total extract. The total extract was loaded on a silica gel CC, eluted with different polar solvents in sequence, and then concentrated to obtain corresponding parts. The methylene chloride fraction and the <italic>n</italic>-butanol fraction were separated and purified by silica gel column chromatography, Sephadex LH-20 gel column chromatography, ODS column chromatography, semi-preparative HPLC, etc. And their structures were identified based on physicochemical properties and various spectroscopic methods. <b>Result::Fourteen compounds were isolated from the dichloromethane fraction and the n-butanol fraction of the <italic>Z</italic>. <italic>bungeanum</italic> and identified as(1<italic>S</italic>, 3<italic>S</italic>)-1-methyl-1, 2, 3, 4-tetrahydro-<italic>β</italic>-carboline-3-carboxylic acid(<bold>1</bold>), (3<italic>S</italic>)-1, 2, 3, 4-tetrahydro-<italic>β</italic>-carboline-3-carboxylic acid(<bold>2</bold>), apigenin-7, 4′-dimethyl ether(<bold>3</bold>), genkwanin(<bold>4</bold>), lcariside F₂(<bold>5</bold>), breyniaionoside A(<bold>6</bold>), 3-methoxyphenethyl alcohol-4-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucopynanoside(<bold>7</bold>), 1-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucopyranosyloxy-3-methoxy-5-hydroxybenzene(<bold>8</bold>), orcinol-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucopyranoside(<bold>9</bold>), syringin(<bold>10</bold>), 4-[(3<italic>S</italic>)-3-hydroxybutyl]-2-methoxyphenyl-<italic>β</italic>-<italic>D</italic>-glucopyranoside(<bold>11</bold>), (+ )-lyoniresinol-3a-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucopyranoside(<bold>12</bold>), 2-methylpropanyl-6-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-apiofuranosyl-<italic>β</italic>-<italic>D</italic>-glucopyranoside(<bold>13</bold>)and(<italic>E</italic>)-6-hydroxy-2, 6-dimethylocta-2, 7-dienoic acid(<bold>14</bold>). <b>Conclusion::All compounds were isolated from <italic>Z</italic>. <italic>bungeanum</italic> for the first time, and compounds <bold>1-4, 12</bold> and <bold>14</bold> were isolated from this genus for the first time.

Chemical Constituents from Stems of Zanthoxylum bungeanum / 中国实验方剂学杂志

Objective:To isolate and identify the chemical constituents from 95%ethanol extracts from stems of Zanthoxylum bungeanum. Method:The stems (25 kg) were extracted with 95%ethanol under reflux for three times and then filtrated,and the combined filtrates were concentrated under vacuum to get the extracts. After suspension with water,the extracts were extracted with petroleum ether,dichloromethane,ethyl acetate and n-butanol successively,and five corresponding parts were obtained. The parts of petroleum ether,dichloromethane,ethyl acetate were separated in a similar way to get pure compounds. By the method the subjects were first chromatographed on silica gel column respectively,then the selected sub-fractions were further separated by Sephadex LH-20, and finally purified on preparation HPLC to get the monomer compound. The n-butanol part was first treated with macroporous resin D101,and then the sub-fractions were further purified by almost the same method as mentioned above. The structures of these compounds were determined by spectrum technology as IR,MS,1H-NMR,13 C-NMR. Result:Ten compounds were isolated from Z. bungeanum stems and identified as follows:(+)-magnoflorine(1),(-)-marmesin(2),(-)-columbianetin(3),(-)-decursinol(4),lupeol(5),α-amyrin(6),β-amyrin(7),δ-amyrin(8),quercetin(9),rutin(10). Among them, 1 was alkaloid, 2,3,4 were coumarin, 5,6,7,8 were triterpenoid, 9,10 were flavonoids. Conclusion:Compounds 1-8 were isolated from this plant for the first time.

Effect and mechanism of essential oil from Zanthoxylum bungeanum in microemulsion gel preparation on percutaneous delivery of complex components / 中国中药杂志

This study aimed to investigate the transdermal enhancing effect of essential oil from Zanthoxylum bungeanum(Z. bungeanum oil) in microemulsion gel(ZO-ME-gel) on permeation of different components,and reveal the transdermal enhancing mechanism of ZO-ME-gel. A series of components with different log P values were selected as model drugs and encapsulated in ZO-ME-gel to simplify and characterize the complex components of traditional Chinese medicine. The transdermal behavior of the model drugs was further examined using the improved Franz diffusion cell method. Then attenuated total reflection Fourier transform infrared spectroscopy(ATR-FTIR),differential scanning calorimetry(DSC) studies and hematoxylin-eosin(HE) staining were used to investigate the effects of Z. bungeanum oil and ZO-ME-gel on keratin,intercellular lipids and microstructure of the stratum corneum(SC). The results showed that Z. bungeanum oil and ZO-ME-gel had a good transdermal enhancing effect on both hydrophilic and lipophilic drugs,and the best effect was achieved when log P value was-0. 5. The transdermal enhancing mechanism of Z. bungeanum oil and ZO-ME-gel was related to affecting the order of the SC lipids,changing lipid fluidity and protein conformation,and disrupting the integrity of the SC structure. 5% Z. bungeanum oil had greater transdermal enhancing effect and destruction of SC structure than ZO-ME-gel. These results suggested that Z. bungeanum oil loaded in microemulsion gel still had a good transdermal enhancing effect although the effect was not as great as Z. bungeanum oil itself,in addition,ZO-ME-gel was less irritating to the skin and safer to use,which had a guiding role in the development and clinical application of Z. bungeanum oil-containing traditional Chinese medicine topical preparations.

Textual research on origin of Zanthoxylum armatum and consideration of its definition / 中草药

As a characteristic medical and edible products, Zanthoxylum armatum has come into being popular in the past 20 years, but it has not been recorded in the relevant standards of Z. bungeanum commodities, and there is no clear regulation for the definition of Z. armatum. This paper makes a textual research on the sources of Z. bungeanum commodities, and analyzes the differences and connections between Z. armatum and other Z. bungeanum commodities, and puts forward the new definition of Z. armatum. The definition of Z. armatum will help to stabilize the quality of Z. armatum, provide reference for the quality standard and quality evaluation of Z. armatum, and provide a reference for solving the mixed situation of Z. schinifolium in Z. bungeanum commodities.

Quality standard for Compound Tougucao Solution / 中成药

AIM To establish the quality standard for Compound Tougucao Solution [Impatiens balsamina L.Zanthoxylum bungeanum Maxim.,Gleditsia sinensis Lam and Momordica cochinchinensis (Lour.) Spreng.].METHODS The fingerprints of ten batches of samples were established,after which TLC was used for the qualitative identification of I.balsamina and Z.bungeanum,HPLC was adopted in the content determination of quercetin.RESULTS There were twenty-seven common peaks in the fingerprints with the similarities of 0.978-0.986,four of which were sodium benzoate,rutin,hyperoside and quercetin,respectively.The clear TLC spots were free from negative interference.Quercetin showed a good linear relationship within the range of 1.68-107.6 μg/mL (R2 =0.999 9),whose average recovery was 104.3％ with the RSD of 2.2％.CONCLUSION This stable and reproducible method can be used for the quality control of Compound Tougucao Solution.

Chemical constituents from Zanthoxylum bungeanum Dahongpao / 中成药

AIM To study the chemical constituents from Zanthoxylum bungeanum Dahongpao.METHODS The 95％ ethanol extract from Z.bungeanum Dahongpao was purified by silica column and Sephadex LH-20,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Nine compounds were isolated and identified as β-sitosterol (1),hydroxy-β-sanshool (2),daucosterol (3),quercetin-3-O-galactoside (4),3-methoxyquercetin (5),succinic acid (6),mannitol (7),arbutin (8),7-O-α-Dglucosyl-3,8-dihydroxy-2-(3-hydroxy-4-methoxyphenyl)-5-methoxy-4H-1-benzopyran-4-one (9).CONCLUSION Compounds 5 and 7 are isolated from genus Zanthoxylum for the first time,compounds 1,3-9 are first isolated from this plant.

Analysis and evaluation of resourceful chemical compositions in different parts of Zanthoxylum bungeanum fruit and its seed oil / 中国中药杂志

The study aims of this study is to analyze and evaluate the resourceful chemical compositions in different parts of mature fruit of Zanthoxylum bungeanum, and provide a scientific basis for the comprehensive utilization of this medicinal plant resources. GC-MS method was used to analyze the volatile oils and the fatty acids, and HPLC method was used to determine the flavonoids and phenolic acids in the pericarp, seed and seed oil of Z.bungeanum. There were 26, 19 and 11 kinds of volatile components detected in the pericarp, seed and seed oil, respectively, in which terpenoids and their oxy-derivatives were the main components, and the contents of linalool and its esters in pericarp were relatively high. The contents of total fatty acids in the pericarp, seed and seed oil were 108.42, 331.63, 966.04 mg•g⁻¹, respectively.Oleic acid, linoleic acid andα-linolenic acid were abundantin all samples. The pericarp contains relatively high content of flavonoids, such as hyperoside, quercitrin, rutin, isoquercitrin, while the above components were not detected in the seed and seed oil. These results confirmed that the fruit of Z.bungeanum contains high contents of the resourceful chemical compositions, and their composition and contents were differed among organs, which provide a scientific basis for the utilization of Z.bungeanumfruit.

The Effect of the Volatile Oil of Zanthoxylum Bungeanum on Human Demodex in vitro / 医学研究杂志

Objective To observe the effect of the volatile oil of zanthoxylum bungeanum in killing demodex in vitro.Method The mites were collected with adhesive cellophane tape technique.The killing effect of the pure volatile oil with different concentrations volatile oils on human demodex was investigated by microscope.Results The pure volatile oil was highly powerful in killing D.f and D.b in vitro,and for the D.b,the killing effect was better.With the increase of dilutus multiple,the killing effect dareased.Conclusion The pure volatile oil of zanthoxylum bungeanum is an effective component in killing demodex in vitro.