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.

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.

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.

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。

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.

基于HPLC指纹图谱和网络药理学的高良姜质量标志物预测分析

目的:将指纹图谱与网络药理学相结合预测分析高良姜的质量标志物(Q-Marker)。方法:采用高效液相色谱法(HPLC)对高良姜进行成分分析,对20批高良姜提取物成分进行相似度评价、聚类分析和主成分分析,筛选高良姜提取物中Q-Marker候选成分;对筛选的Q-Marker候选成分进行靶点收集和网络药理学分析,构建“成分-靶点-通路”网络,并预测高良姜提取物的Q-Marker。结果:20批高良姜指纹图谱相似度集中在0.952~1.000范围,提示不同产地和批次的高良姜提取物整体组成上具有一致性;聚类分析和主成分分析显示海南的高良姜样品聚类趋势比较一致,云南、广东和广西的高良姜未按产地归类,不同产地的高良姜在成分上存在着差异,说明产地只是高良姜质量的影响因素之一。根据HPLC共有成分以及聚类分析和主要成分分析,得到2个Q-Marker候选成分,分别为高良姜素和槲皮素。经网络药理学分析筛选6个关键活性成分,6个关键要点和21条通路。结论:本研究初步预测槲皮素、山奈酚和高良姜素为高良姜的Q-Marker,为高良姜的质量控制予与参考,同时也为高良姜功效关联物质的研究及作用机制的阐释奠定了基础。

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.

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.

高良姜的盐炙工艺及其盐炙前后HPLC指纹图谱对比研究

目的:优选盐炙高良姜的炮制工艺,并对比研究其盐炙前后的HPLC指纹图谱变化。方法:以外观性状、含水量、水溶性浸出物百分含量及高良姜素含量为评价指标,采用单因素试验,以切片厚度、炒制温度和炒制时间为指标,设计正交试验优选盐炙高良姜的炮制工艺。采用HPLC法测定盐炙前后高良姜样品的高良姜素含量和HPLC指纹图谱。采用“中药色谱指纹图谱相似度评价系统(2004 A版)”进行相似度评价,并应用SIMCA_P+12.0软件进行主成分分析。结果:优选的盐炙高良姜最佳炮制工艺为用5%食盐水浸润至药透汁尽,然后120℃炒制6 min;盐炙后的高良姜样品中高良姜素含量与生品比较,差异无统计学意义(P>0.05);盐炙后的高良姜样品中水溶性浸出物百分含量均高于生品(P<0.01);盐炙后的高良姜化学成分种类少于生品;虽然它们的指纹图谱相似度较高,但以其指纹图谱峰面积为变量进行OPLS-DA分析可以快速地将盐炙前后的高良姜样品区别开。结论:优选的盐炙高良姜炮制工艺稳定可行,可为生产盐炙高良姜饮片提供实验依据;盐炙前后的高良姜饮片内在化学成分发生了变化,这从化学成分的角度揭示了盐炙高良姜的炮制机理。

高良姜中总黄酮提取与DPPH自由基清除活性研究

通过正交试验优化了高良姜黄酮的提取工艺。结果表明,乙醇浸提法提取高良姜总黄酮的最佳工艺条件为浸提温度70℃、料液比1∶25g/mL、乙醇浓度60%、提取时间2h。通过DPPH自由基清除实验研究了高良姜总黄酮的抗氧化活性,结果表明:高良姜黄酮具有一定的抗氧化活性,且随着浓度增加,抗氧化性能增强。

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。

高良姜饮片HPLC指纹图谱及高良姜素含量测定研究

目的:采用高效液相色谱法建立高良姜饮片的HPLC指纹图谱,同时测定样品中高良姜素的含量,为其质量控制提供新方法。方法:以Phenom enex C18柱(4.6 mm×250 mm,5μm)为色谱柱,流动相为乙腈-0.2%磷酸溶剂系统,线性梯度洗脱,检测波长为280 nm(高良姜素含量测定为235 nm);采用指纹图谱相似度软件进行数据分析。结果:确定了高良姜饮片9个共有峰,建立了14批药材的共有图谱,并比较了样品中高良姜素的含量。结论:指纹图谱信息结合有效成分含量测定,可较为全面评价高良姜饮片的质量。

HPLC-DAD测定高良姜药材中山奈素的含量

目的测定不同产地高良姜药材中山奈素的含量。方法使用HPLC-DAD分析方法,并进行了方法学的考察。结果山奈素的线性范围在0.02～0.11μg时,呈良好的线性,r=0.9997。结论药材中的主要峰均达到了有效的分离性,其山奈素的含量范围为0～0.2140mg.g-1。

HPLC法测定高良姜中原儿茶酸的含量

目的建立高良姜中原儿茶酸的含量测定方法。方法采用HPLC法。色谱柱为D iamonsilODS C18柱（200 mm×4.6 mm,5μm）,流动相为甲醇-质量分数为0.05%的磷酸水溶液（体积比为12∶88）,流速为1.0 mL.min^-1,检测波长为258 nm,柱温为35℃,进样量为20μL。结果原儿茶酸在0.902-14.432 mg.L^-1内与峰面积呈良好的线性关系,平均回收率为99.2%,RSD=1.8%（n=9）。结论该方法可为高良姜药材质量控制提供新指标。

HPLC测定高良姜中槲皮素 山柰素和异鼠李素的含量

目的：建立高良姜中槲皮素、山柰素和异鼠李素的含量测定方法。方法：采用HPLC法。Kromasil C18色谱柱（5μm,4.6mm×250mm）,流动相：甲醇-0.4%磷酸溶液（52：48）,检测波长：360nm。结果：槲皮素浓度线性范围0.962-24.05μg/mL,r=0.99994;山柰素浓度线性范围1.008-25.2μg/mL,r=0.99990;异鼠李素浓度线性范围0.983-24.575μg/mL,r=0.99996,样品平均回收率为槲皮素97.97%,RSD=1.59%（n=6）;山柰素97.35%,RSD=1.86%（n=6）;异鼠李素98.11%,RSD=2.37%（n=6）。结论：该方法简便、准确、重现性好,可用于该中药的含量测定。

基于“敲除策略”筛选高良姜乙酸乙酯部位抗幽门螺杆菌胃炎活性成分

目的筛选高良姜乙酸乙酯部位抗幽门螺杆菌胃炎活性成分。方法联合“敲除策略”与高效液相色谱(HPLC)检测对高良姜乙酸乙酯部位的组分(成分)进行分离,同时得到不含该组分(成分)的阴性样品;构建幽门螺杆菌感染人胃黏膜上皮细胞(GES-1)胃炎模型,采用酶联免疫吸附测定(ELISA)法检测细胞上清液中白细胞介素(IL)-6、肿瘤坏死因子-α(TNF-α)、IL-8和IL-1β水平。结果总黄酮组分、不含二苯基庚烷的阴性组分、不含5-羟基-7-(4-羟基-3-甲氧基)苯基-1-苯基-3-庚酮(DHPA)的阴性组分以及高良姜素(给药浓度8μg·mL^(-1)作用24 h),均能够显著降低幽门螺杆菌胃炎细胞上清液中IL-6水平;总黄酮组分浓度为16μg·mL^(-1)时可显著抑制幽门螺杆菌感染GES-1胃炎细胞IL-6、TNF-α、IL-8和IL-1β的释放。结论总黄酮组分是高良姜乙酸乙酯部位抗幽门螺杆菌胃炎的主要活性组分,该研究结果为进一步阐释高良姜抗幽门螺杆菌胃炎药效物质基础奠定了基础。

高良姜多糖的提取及其活性研究

采用水提醇沉和单因素实验获得较优的高良姜多糖提取方法。以对DPPH·自由基、超氧阴离子自由基的清除率为指标,评价高良姜多糖的抗氧化能力,同时研究其对酪氨酸酶抑制以及抗菌活性,探讨其在护肤品领域的潜在应用。获得高良姜多糖较优的提取方法为:提取温度为70℃、提取时间为40 min、料液比为1∶40(g/mL),此时多糖得率为11.62%,含量为20.0%。同时,研究发现高良姜多糖对DPPH·自由基和超氧阴离子自由基具有较强的清除率,均表现出质量浓度依赖性;研究发现高良姜多糖对酪氨酸酶具有较强的抑制活性,IC50值为2.69 mg/mL;此外,研究发现高良姜多糖对白色念珠菌和金黄色葡萄球菌具有较强的抑制作用。结果表明,高良姜多糖在护肤品方面具有较好的应用前景。