Study on mechanism of two related Chinese herbs A.officinarum-Pogostemon in treatment of delayed emptying in diabetic gastroparesis based on network pharmacology

Objective:To explore the potential active components,therapeutic targets and critical path of Alpinia officinarum and Pogostemonis Herba in the treatment of diabetic gastroparesis(DGP)by using network pharmacology.Methods:The main chemical components and corresponding targets genes of A.officinarum-Pogostemonis Herba were screened through TCMSP database retrieval[oral bioavailability(OB)≥30%and drug like(DL)≥0.18].Tgenes of diabetic gastroparesis were screened by the Human Gene Database(GeneCards),and Venny 2.1 software was used to obtained common targets for the active ingredients of A.officinarum-Pogostemonis Herba and DGP.Then,the protein-protein interaction(PPI)network of the common targets was constructed by STRING database and analyzed to performed the core targets.GO function and KEGG pathway enrichment analysis of the common target genes were obtained by using ClusterProfiler R package.Finally,the network diagram of"active ingredient-pathway-target"was used to establish by Cytoscape 3.8 software.Results:Totally 23 ingredients of A.officinarum-Pogostemonis Herba,97 active ingredients targets and 533 DGP related targets,including 46 common targets were selected.The common targets were mainly enriched in the cell constituents such as the nuclear chromatin and mitochondria outer membrane,involved in the biological processes as oxidative stress,apoptosis signal regulation,and molecular functions as enzyme binding,protein phosphatase binding,and cytokine activity.They were also concentrated in the signal pathways such as PI3K/Akt,HIF-1 and MAPK.The network of“active ingredients-targets-pathways”indicated the active components such as quercetin,kaempferol and galangin in A.officinarum-Pogostemonis Herba played an anti-delayed gastric emptying in diabetic gastroparesis by acting on PTGS2,NOS2,BCL2,IL6,VEGFA and other targets to jointly regulate PI3K-Akt,HIF-1 and MAPK pathways.Conclusion:This study initially reveals that the combined treatment of A.officinarum-Pogostemonis Herba for delayed gastric emptying in diabetic gastroparesis is a complex process with multi-components,multi-targets and multi-pathways,and provides a new idea for followup researches.

甘蔗(Saccharum officinarum L.)品种遗传差异的AFLP分子标记分析

采用AFLP技术 ,从 6 4对引物中筛选出 5对带型分布均匀、多态性丰富且分辨能力强的引物 ,对两个在生产上广泛推广应用的甘蔗品种的AFLP指纹图谱进行分析 ,计算了两品种间的遗传相似性和遗传距离。结果表明 ,5对引物在 2个甘蔗品种间均存在显著的差异 ,其中多态性位点占总扩增位点的 10 2 % ,区分率达 10 0 %。这对应用AFLP技术鉴定甘蔗品种的真伪提供了依据。本文还对甘蔗AFLP分析体系的关键技术以及银染的安全性和可靠性进行了讨论。

Effect of Alpinia officinarum Hance alcohol extracts on primary dysmenorrheal

Objective:To study the effect of Alpinia officinarum Hance(A.officinarum) 80% alcohol extract on the primary dysmenorrhea.Methods:A.officinarum 80% alcohol extract were enriched by macroporous adsorption resins.Female mice of primary dysmenorrhea model were established by oxytocin induction; the effects of A.officinarum 80% alcohol extract on primary dysmenorrhea were observed by body twist method; and the homogenate level of prostaglandin F_(2α)(PGF_(2α)),prostaglandin E_2(PGE_2) and Ca^(2+) in the uterus were observed in oxytocin-induced female mice.Results:The writhing frequency of primary dysmenorrhea mice was significantly decreased after treatment of A.officinarum 80% alcohol extract and the level of PGF_(2α),PGE_2 and Ca^(2+) in mice uterus was significantly decreased(P<0.05,P<0.01) in groups of mice treated with middle and high dosage of A.officinarum 80% alcohol extract compared with that of model group.Conclusions:These findings suggest that A.Officinarum 80% alcohol extract can significantly relieve primary dysmenorrhea.

Radiation-induced in vitro mutagenesis system for salt tolerance and other agronomic characters in sugarcane(Saccharum officinarum L.)

Gamma ray-induced in vitro mutagenesis and selection for salt(NaC l) tolerance were investigated in sugarcane(Saccharum officinarum L.). Embryogenic callus cultures were irradiated(10 to 80 Gy) and subjected to in vitro selection by exposure of irradiated callus to NaC l(0, 50, 100,150, 200, and 250 mmol L-1). Increasing NaC l concentrations resulted in growth reduction and increased membrane damage. Salt-selected callus lines were characterized by the accumulation of proline, glycine betaine, and Na+and K+concentration. Higher accumulation of proline and glycine betaine was observed in NaC l stressed callus irradiated at 20 Gy. Na+concentration increased and K+concentration decreased with increasing salt level. Irradiated callus showed50–60% regeneration under NaC l stress, and in vitro-regenerated plants were acclimatized in the greenhouse, with 80–85% survival. A total of 138 irradiated and salt-selected selections were grown to maturity and their agronomic performance was evaluated under normal and saline conditions. Of these, 18 mutant clones were characterized for different agro-morphological characters and some of the mutant clones exhibited improved sugar yield with increased Brix%,number of millable canes, and yield. The result suggest that radiation-induced mutagenesis offers an effective way to enhance genetic variation in sugarcane.

Anti-inflammatory activity of Alpinia officinarum hance on rat colon inflammation and tissue damage in DSS induced acute and chronic colitis models

The purpose of this study was to investigate the possible prophylactic effects of Alpinia officinarum hance on experimentally induced acute and chronic colitis models,in-vivo and in-vitro.Acute and chronic colitis were induced in Male Wistar rats by administration of Dextran Sulfate Sodium(DSS)in drinking water.DSS induction exhibited colon shrinkage,increased the Disease Activity Index(DAI)score,increased the levels of inflammatory markers and caused severe anemia.DSS induced animals,co-treated with the hexane extract of Alpinia officinarum(HEAO)(200 mg/kg body wt),effectively suppressed colonic injury that was evidenced by the reduced DAI score,colon weight/length ratio,histological damage,proinflammatory markers and MPO activity.Further,it restored the colonic antioxidants near to normal levels by regulating the oxidative stress via attenuation of lipid peroxidation.Our results revealed that the degree of colitis caused by the administration of DSS was significantly attenuated by HEAO.In addition,the in-vitro study showed that HEAO treatment inhibited the proliferation of HT-29 cells and down regulated the mRNA expression of NF-B and COX-2.Taken together,these results suggest that HEAO is a promising anti-oxidant and anti-inflammatory agent that support its possible therapeutic role in the treatment of colitis.

Implementation and Evaluation of Certain Properties of a Polymer Matrix Composite Material Reinforced by Fibrous Residues of <i>Saccharum officinarum</i>in View of an Applicability Orientation

This study examines the implementation and characterization of a polymer matrix composite material reinforced by sugarcane residues. The aim of the study is to enhance the abundantly produced sugarcane bagasse in the form of residues in the processing plants of said sugar cane. The composite material developed takes into account the size parameters and mass load rate of the reinforcement, the variations of which are between 2.5 mm and 4 mm respectively for the first parameter and 10% to 25% for the second. The load on the test tube during the test has a random orientation. The composite is polyester matrix. The cold compression moulding technique was used in the production of the various samples. Physical properties such as the rate of water absorption and the density of the composite are assessed. The three-point bending tests are carried out with the aim of inducing Young’s modulus from the elaborate samples. Analysis of physical properties shows that water absorption increases with the rate of residue load. The average water absorption rate of bagasse fibres is estimated at 8% for the 2.5 mm fiber size and 12% for the 4 mm fiber size for all composites. Mechanical characterization by bending tests reveals a fragile behavior of the samples tested. Young’s modulus decreases when the load rate of fibrous residues increases regardless of the size of the fibers.

Alpinia officinarum Hance extract alleviates particulate matter-induced lung injury in mice

Objective: To evaluate the effect of Alpinia officinarum Hance(A. officinarum) extract on lung injury caused by particulate matter(PM). Methods: The Kunming mice were intranasally instilled with PM and treated with A. officinarum extract for 3 weeks. Bronchoalveolar lavage fluid, blood and lung samples were collected for biochemical, serological and histopathological studies. Results: Serological analysis showed that albumin levels, lactate dehydrogenase and alkaline phosphatase activities in bronchoalveolar lavage fluid were significantly reduced after administrations of 50, 100 and 200 mg/kg of A. officinarum extracts to the PM injured mice. Markers of oxidative stress, nitric oxide, malondialdehyde levels and nitric oxide synthase activities, were significantly decreased. Correspondingly, total superoxide dismutase activity was improved dramatically. The expressions of interleukin-6 and tumor necrosis factor alpha were also down-regulated obviously. In addition, pathological sections of lung tissue showed that A. officinarum could reduce the infiltration of inflammatory cells, pulmonary edema and pulmonary fibrosis. These results showed that A. officinarum extract could alleviate PMinduced lung injury via reducing the permeability of cell membranes in lung tissue, eliminating oxidative stress and relieving inflammatory response.Conclusions: A. officinarum extract was an efficient treatment for PM-induced lung injury in mice, and it may be a promising therapeutic agent in future.

Herb-drug interaction in the protective effect of Alpinia officinarum against gastric injury induced by indomethacin based on pharmacokinetic, tissue distribution and excretion studies in rats

Alpinia officinarum Hance of the Chinese traditional herb for the treatment of emesis, abdominal pain and diarrhea has been used to counteract gastric disease induced by indomethacin in rats without obvious side effects. However, the role of herb-drug interaction between indomethacin and A. officinarum based on pharmacokinetic, tissue distribution and excretion still remains unknown. In this study, an ultra-fast liquid-tandem mass spectrometry(UFLC-MS/MS) method was developed for simultaneous determination of indomethacin and its three metabolites, O-desmethylindomethacin(ODI), deschlorobenzoylindomethacin(NDI) and indomethacin acyl-b-D-glucuronide(IDAbG) by oral administration of indomethacin solution with and without the ethanolic extract of A. officinarum and applied to comparative pharmacokinetic, tissue distribution and excretion studies. Our results clarified that oral administration of A. officinarum produced significant alterations in the pharmacokinetic parameters of indomethacin. And the pharmacokinetic interaction between indomethacin and A. officinarum reduced the systemic exposure of indomethacin and increased its elimination. Tissue distribution results demonstrated that co-administration of A. Officinarum could not reduce the accumulation of indomethacin in the target tissue of the stomach, but could accelerate the excretions of indomethacin and its three metabolites including ODI, NDI and IDAb G in the bile and feces of rats in the excretion study.Therefore, A. Officinarum might have a gastrointestinal protective effect through the interaction role with indomethacin based on the pharmacokinetics and excretion in rats.

The effect of characteristic Li-medicine Alpinia officinarum Hance on improving insulin resistance

Objective:To study the effects of characteristic Li-medicine Alpinia officinarum Hance on improving insulin resistance(IR),and provide scientific evidence for the adjuvant treatment T2DM.Methods:CCK8 kit was used to detect the viability of HepG2 cells with 0-200μg/mL Alpinia officinarum Hance extract(AOE)and 0-100μmol/L galangin,and determine the administration concentration.The IR model was established by inducing HepG2 cells with high glucose for 48 hours,and rosiglitazone(ROSI)was used as the positive control drug.Furthermore,we detected intracellular and extracellular glucose contents in IR-HepG2 cells after dosing AOE and galangin by flow cytometry and glucose detection kit,and evaluated the effect of regulating glucose metabolism and IR.Western blot was used to detect the effects of AOE and galangin on the expression of glucose transporter-4(GLUT4)in IR-HepG2 cells.Results:CCK8 kit test result showed that 0-200μg/mL AOE and 0-50μmol/L galangin had no significant effect on HepG2 cell viability.The results of flow cytometry and glucose detection kit showed that the glucose uptake and consumption of the IR model group induced by high glucose was significantly lower than that of the blank group,and thus the IR model was successfully established.Both 50μg/mL AOE and 10,20μmol/L galangin significantly increased the glucose uptake and consumption of IR-HepG2 cells(P<0.05);Western blot experiment showed that AOE and galangin significantly increased GLUT4 level of IR-HepG2 cells(P<0.01).Conclusion:Alpinia officinarum Hance could promote the uptake and consumption of glucose in IR-HepG2 cells by up-regulating the expression of GLUT4 protein,thereby improving IR and enhancing insulin sensitivity.

Hepatic Impact of Mandragora officinarum Leaf Extract on Wistar Albino Rats

This study evaluates the hepatic effects of Mandragora officinarum leaf extract on wistar albino rats. A total of twenty-four (24) rats were randomly divided into 4 groups labeled A, B, C and D and kept in a well-ventilated room. Group A served as control and these rats were fed distilled water. Rats in groups B, C, and D were given three (3) different doses of the leaf extract (1.5, 3.5 and 5.0 mL/KgBW) respectively. They were administered once daily for 14 and 28 days consecutively. Animals were sacrificed 24 hours after the last treatment. Blood samples were collected into heparinized sample bottles for analysis. Aspartate aminotransferase, Alanine aminotransferase and histology results were normal when the leaf extract was given for 14 days. Alkaline phosphatase significantly decreased within the same time interval. Alkaline phosphatase increased in a dose-dependent manner for 28 days. All other liver enzymes were within normal limits. Histopathological changes were seen in all doses when Mandragora officinarum leaf extract was used for 28 days. These changes also worsened with increasing doses. This suggests that the use of mandragora officinarum leaf extract for long periods at a time can cause hepatic damage.

Chemical Composition, Proapoptotic and Antiosteoporosis Activities of the Essential Oil from the Aerial Part of <i>Alpinia officinarum</i>Hance

Background: Alpinia officinarum Hance is valued as an edible medicinal plant. The rhizome is widely reported to have anticancer activity whereas little information is available on the aerial part. This study investigates chemical composition, proapoptotic and anti-osteoporosis activities of essential oil from aerial parts of A. officinarum (APEO). Methods: In this study, APEO was extracted by hydrodistillation and analyzed using GC-MS. The inhibitive activity of 0 - 2.5 μL/mL. APEO was investigated using MTT assay, while in vivo effect was evaluated in nude mice. The cell cycle, apoptosis, Δψm and expression of proteins analyses influenced by 0 - 0.313 μL/mL APEO were detected by PI, Annexin V/PI, JC-1, and Western blot, respectively. Alkaline phosphatase activity and mineralized nodules formation of rat osteoblasts with 0 - 0.156 μL/mL APEO were assayed using colorimetric method and alizarin red staining, respectively. Results: Total 45 constituents were identified accounting for 91.1% of APEO (sesquiterpene hydrocarbons for 44.4%). APEO significantly inhibited cancer cells growth in a dose-dependent manner. APEO also inhibited cancer growth in vivo. The percentage of S phase cells is up to 64.846% with 0.313 μL/mL APEO. The proportion of total apoptotic cells significantly increased to 79.6% at 0.313 μL/mL concentration. APEO treated cells accompanied with Bcl-2 and Δψm decrease, and caspase-3 and p53 upregulation. Furthermore, addition of APEO in rat osteoblasts led to a dose-dependent increase in ALP activity and formation of mineralized bone nodules. Conclusions: Our data suggest that APEO could be developed as an agent against human lung cancer and osteoporosis, especially cancer-induced bone loss.

Genetic analysis for brix weight per stool and its component traits in sugarcane (Saccharum officinarum)

Brix weight per stool (BW) of sugarcane is a complex trait, which is the final product of a combination of many components. Diallel cross experiments were conducted during a period of two years for BW and its five component traits, in- cluding stalk diameter (SD), stalk length (SL), stalk number (SN), stalk weight (SW), and brix scale (BS) of sugarcane. Phenotypic data of all the six traits were analyzed by mixed linear model and their phenotype variances were portioned into additive (A), dominance (D), additive×environment interaction (AE) and dominance×environment interaction (DE) effects, and the correlations of A, D, AE and DE effects between BW and its components were estimated. Conditional analysis was employed to investigate the contribution of the components traits to the variances of A, D, AE and DE effects of BW. It was observed that the heritabilities of BW were significantly attributed to A, D and DE by 23.9%, 30.9% and 28.5%, respectively. The variance of A effect for BW was significantly affected by SL, SN and BS by 25.3%, 93.7% and 17.4%, respectively. The variances of D and DE effects for BW were also significantly influenced by all the five components by 5.1%～85.5%. These determinants might be helpful in sugarcane breeding and provide valuable information for multiple-trait improvement of BW.

高良姜-广藿香配伍溶液的质量标准研究

目的:建立高良姜-广藿香配伍溶液的质量标准。方法:针对高良姜-广藿香配伍溶液,高良姜素、广藿香酮2种指标性成分的薄层鉴别方法分别为:薄层板为硅胶G薄层板和高效硅胶G薄层板,展开剂为石油醚-乙酸乙酯(5∶3)、(5∶1);高良姜素、广藿香酮2种指标性成分高效液相色谱含量检测方法为:色谱柱为C18色谱柱(4.6 mm×250 mm,5μm)、流动相为甲醇(A)-0.2%磷酸(B)、检测波长:310 nm、梯度洗脱。结果:对于高良姜-广藿香配伍溶液中高良姜素、广藿香酮2种指标性成分,薄层鉴别主斑点清晰、阴性对照无干扰;2种指标性成分的浓度在10.00~1000.00μg/mL、10.00~640.00μg/mL范围内与峰面积线性关系系数分别为R^(2)=0.9990、R^(2)=0.9999;精密度RSD分别为0.63%、0.56%,稳定性RSD分别为1.82%、0.24%,重复性RSD分别为1.30%、0.89%;平均回收率分别为103.19%、100.87%(RSD为别为0.93%、0.4%,n=9);该2种指标性成分的含量应分别不低于233.71μg/mL、61.48μg/mL。结论:建立的方法可用于高良姜-广藿香配伍溶液的质量控制,该方法精密度、准确度、稳定性及重复性均符合要求。

Expression of the Grifola frondosa Trehalose Synthase Gene and Improvement of Drought-Tolerance in Sugarcane （Saccharum officinarum L.）

Trehalose Is a nonreduclng dlsaccharlde of glucose that functions as a protectant In the stabilization of blologlcal structures and enhances stress tolerance to abiotic stresses in organisms. We report here the expression of a Grlfola frondosa trehalose synthase （TSase） gene for Improving drought tolerance In sugarcane （Saccharum offlclnarum L.）. The expression of the transgene was under the control of two tandem copies of the CaMV35S promoter and transferred Into sugarcane by Agrobacterium tumefaciens EHA105. The transgenlc plants accumulated high levels of trehalose, up to 8.805-12.863 mg/g fresh weight, whereas It was present at undetectable level in nontransgenlc plants. It has been reported that transgenlc plants transformed with Escherlchla coil TPS （trehalose-6-phosphatesynthase） and/or TPP （trehalose-6-phosphate phosphatase） are severely stunted and have root morphologlc alterations. Interestingly, our transgenlc sugarcane plants had no obvious morphological changes and no growth Inhibition in the field. Trehalose accumulation in 35S-35S：TSase plants resulted In In- creased drought tolerance, as shown by the drought and the drought physiological Indexes, such as the rate of bound water/free water, plasma membrane permeability, malondlaldehyde content, chlorophyll a and b contents, and activity of SOD and POD of the excised leaves. These results suggest that transgenlc plants transformed with the TSase gene can accumulate high levels of trehalose and have enhanced tolerance to drought.

Research and application of Alpinia officinarum in medicinal field

Alpinia officinarum is a medicinal plant and food. Its dried rhizome has been widely used for the relief of symptoms such as stomach aches, colds, ulcer and diarrhea for hundreds of years. Recent pharmacological studies showed it has anti-inflammatory, anti-oxidative, antidiabetic, anti-ulcer, anti-diarrhea, antiemetic,analgesia, anticoagulation, antitumor and antivitiligo effects. In this study, we summarized the current knowledge about its botanical resources, ethanopharmacological function, chemical constituents, pharmacologies and pharmacokinetics, safety and toxicity, and clinical application.

A new dimeric diarylheptanoid from the rhizomes of Alpinia officinarum

AIM: To study the chemical constituents of the rhizomes of Alpinia officinarum Hance. METHOD: Compounds were isolated by repeated column chromatography, and their structures were elucidated on the basis of spectral analysis. The cytotoxic activities of these compounds were evaluated with the T98G and B16F10 cell lines by the MTT assay. RESULTS: A dimeric diarylheptanoid, named alpinin B(1), along with three known diarylheptanoids were obtained, and their structures were identified as alpinin B(1), 1, 7-diphenyl-3,5-heptanedione(2),(4E)-1, 7-diphenylhept-4-en-3-one(3) and(4E)-7-(4-hydroxyphenyl)-1-phenylhept-4-en-3-one(4). CONCLUSION: Compound 1 is a new dimeric diarylheptanoid. The biosynthetic pathway of 1 was speculated to originate from a Michael reaction between compounds 2 and 3. Compound 3 showed cytotoxicity against the human glioblastoma T98G cell line with IC50 of 27 μmol L 1.

基于Keap1/Nrf2/ARE信号通路探讨高良姜总黄酮对铅诱导HK-2细胞损伤的保护作用

目的 探讨高良姜总黄酮对铅(Pb)诱导人肾皮质近曲小管上皮细胞(HK-2)细胞损伤的保护作用。方法 体外培养HK-2细胞,MTT法检测不同质量浓度高良姜总黄酮和Pb对HK-2细胞活力的影响。设置对照组、高良姜总黄酮对照组、模型组和高良姜总黄酮组,除对照组和高良姜总黄酮对照组外各组均给予200μmol/L Pb诱导细胞损伤,同时高良姜总黄酮对照组和高良姜总黄酮组给予100μg/mL高良姜总黄酮干预24 h。通过Hoechst 33258荧光染色法联合annexin V-FITC/PI双标记流式细胞术检测细胞凋亡情况,荧光显微镜法观察Pb诱导及高良姜总黄酮干预对细胞内ROS水平的影响,试剂盒法检测细胞内氧化应激指标ROS、MDA、GSH水平和GSH-Px、CAT、SOD活性,ELISA法检测细胞培养上清液IL-1β、IL-6、TNF-α水平,Western bolt法检测细胞Keap1/Nrf2/HO-1信号通路相关蛋白及caspase-3蛋白表达。结果 与对照组比较,高良姜总黄酮(12.5~200μg/mL)对HK-2细胞活力没有显著影响。与模型组比较,高良姜总黄酮可减少Pb诱导HK-2细胞的凋亡(P<0.05),减轻细胞皱缩等细胞凋亡形态学变化,上调细胞内SOD、CAT、GSH-Px活性及GSH水平(P<0.05),降低细胞内MDA、ROS水平(P<0.05),上调Keap1/Nrf2/HO-1信号通路相关蛋白及caspase-3蛋白表达(P<0.05)。结论 高良姜总黄酮可能通过调控Keap1/Nrf2/ARE信号通路相关蛋白表达,对Pb诱导HK-2细胞损伤起保护作用。

A novel dimeric diarylheptanoid from the rhizomes of Alpinia officinarum

A novel dimeric diarylheptanoid,named alpinin A（1）,along with two known compounds,1,7-diphenyl-5-ol-3-heptanone and 7- （4＇-hydroxy-3＇-methoxyphenyl）-l-phenyl-4-heptene-3-one,were isolated from the rhizomes of Alpinia officinarum Hance.The structure of compound 1 was elucidated on the basis of spectral analysis,including HR-IT-TOF-MS,1D and 2D NMR.And a possible biogenetic pathway had been proposed for 1.