• <tr id="yyy80"></tr>
  • <sup id="yyy80"></sup>
  • <tfoot id="yyy80"><noscript id="yyy80"></noscript></tfoot>
  • 99热精品在线国产_美女午夜性视频免费_国产精品国产高清国产av_av欧美777_自拍偷自拍亚洲精品老妇_亚洲熟女精品中文字幕_www日本黄色视频网_国产精品野战在线观看 ?

    A Network Pharmacology Study on the Effects of Ma Xing Shi Gan Decoction on Influenza

    2020-11-03 03:12:08LONGXiLIULePingXUXinYiLILingZHANGGuoMin
    Digital Chinese Medicine 2020年3期
    關(guān)鍵詞:傷寒論麻黃石膏

    LONG Xi,LIU Le-Ping,XU Xin-Yi,LI Ling,ZHANG Guo-Min*

    a.Hunan University of Chinese Medicine,Changsha,Hunan 410208,China

    b.Department of Blood Transfusion,the Third Xiangya Hospital of Central South University,Changsha,Hunan 410013,China

    Keywords

    Network pharmacology

    Ma Xing Shi Gan Decoction(MXSGD)

    Influenza

    Cytoscape

    iGEMDOCK

    RELA-Licochalcone A

    ABSTRACT

    Objective Pharmacological methods were used to screen targets and signaling pathways of Ma Xing Shi Gan Decoction(MXSGD)during influenza treatments,and mechanisms underlying antiinfluenza effects were elucidated.

    Methods The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and relevant literature were searched under predefined conditions to identify the main compounds and their targets.Interactions between the target proteins were predicted using the STRING database.Gene Ontology(GO)functional enrichment analyses and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses were performed on the core targets involved in the influenza protein-protein interaction(PPI)network,using WebGestalt and the reactome database.iGEMDOCK was used for molecular docking of receptors and ligands to produce docking scores,and the results were visualized using Autodock and PyMOL.

    Results In total,126 major compounds and their respective targets were screened.355 influenza target proteins and 1 221 influenza protein interactions were predicted using the STRING database.Influenza-related signaling pathways were strongly enriched in pharmacodynamic targets of MXSGD such as cytokine signaling in immune system and signaling by interleukin.The main biological process was response to the stimulates.Molecular docking results showed that RELALicochalcone A docking elicited by MXSGD,was superior to that of other target proteins and active compounds,suggesting that the docking site is also the main effector site of MXSGD during influenza treatments.

    Conclusions The results showed that MXSGD exerts antiinfluenza effects by interfering with virus adsorption,inhibiting virus proliferation,influencing immune functions and protecting host cells,which may prevent inflammation-induced tissue damage.

    1 Introduction

    Influenza is a globally occurring infectious disease.After infection,patients frequently suffer from fever,headache,myalgia and general discomfort and can develop pneumonia,which is likely to cause complications[1].In severe cases,influenza can cause multiple organ failure and death.The main subtypes of influenza virus detected in winter and spring 2018 and 2019 belonged to H1N1[2].Influenza vaccination for prevention and treatment is an effective method,but there are still some shortcomings.High pathogenicity,high lethality and low cure rate of influenza can cause substantial damage to affected individuals[3-5].

    According to the theory of traditional Chinese medicine(TCM),the symptoms of influenza are caused by exogenous wind heat and poisonous evil which is trapped in the lungs.The basic pathogenesis comprises evil heat,silicosis and obstruction of the respiratory tract[6].Chinese medicine suggests “pattern identification”,thus treatment of influenza should include diffusing the lung.The textTreatise on Cold Damage Diseases(Shang Han Lun,《傷寒論》)notes “After sweating,you can’t make Gui Zhi Decoction.If you sweat and asthma without heating,you can use Ma Xing Shi Gan Decoction(MXSGD).” MXSGD is used to treat influenza,and it consists of four herbal medicines:Ephedra Herba(Ma Huang,麻黃),Armeniacae Semen Amarum(Ku Xing Ren,苦杏仁),Glycyrrhizae Radix Et Rhizoma(Gan Cao,甘草)and Gypsum Fibrosum(Shi Gao,石膏).It has similar effects as diffusing the lung and may help treat cough and asthma symptoms of pulmonary inflammation.A vast body of literature suggests that MXSGD can exert positive effects in influenza treatments[7-12].

    TCM may offer considerable advantages regarding treatments of complex diseases,however,the current lack of research methods for investigating diseases and active compounds of TCM may prevent scientific assessment of its systematic application.Network pharmacology based on disease-target-drug interaction networks has been used previously to examine relationships between multi-compound and multi-target systems of TCM and to explore mechanisms of human disease and health[13].This research method incorporates a holistic and systematic approach,and it agrees with the basic theory of TCM,the overall concept of TCM,and with prescription treatment; moreover,this method can explain the mechanism of “regular medicine-medicine” affecting diseases[14,15].Beneficial effects of MXSGD in influenza treatments have not been sufficiently elucidated due to the complexity of working mechanisms of active compounds of prescription drugs and interactions of influenza virus targets.The objective of the present study was to explore the mechanisms of influenza treatments based on MXSGD through network pharmacology and molecular docking.

    2 Materials and Methods

    2.1 Collection and screening of MXSGD components and corresponding targets

    We used the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP,http://lsp.nwu.edu.cn/tcmsp.php)[16]with the search terms “Ephedra Herba(Ma Huang,麻黃)”“Armeniacae Semen Amarum(Ku Xing Ren,苦杏仁)” and “Glycyrrhizae Radix Et Rhizoma(Gan Cao,甘草)” to account for all compounds of MXSGD.The compounds of Gypsum Fibrosum(Shi Gao,石膏)was not found,thus we predominantly relied on literature research to identify the main chemical components of Gypsum Fibrosum(Shi Gao,石膏).Compounds with high activity according to TCMSP were produced by screening the compounds of Ephedra Herba(Ma Huang,麻黃),Armeniacae Semen Amarum(Ku Xing Ren,苦杏仁)and Glycyrrhizae Radix Et Rhizoma(Gan Cao,甘草)with oral bioavailability(OB)≥ 30% and drug-like(DL)≥0.18.The single main compound of Gypsum Fibrosum(Shi Gao,石膏)was not screened.Through the TCMSP platform,the selected compounds were collected in respective target databases to relate to their targets.The target of Gypsum Fibrosum(Shi Gao,石膏)was retrieved using the STITCH platform(http://stitch.embl.de).

    2.2 Influenza disease target collection

    Using the Comparative Toxicogenomics Database(CTD)[17](http://ctd.mdibl.org/)and the Online Mendelian Inheritance in Man(OMIM)database[18](https://www.omim.org/),targets of influenza viruses were compiled using the search term“influenza”.In total,7 790 targets were produced,and all targets identified in the OMIM database were selected,as the amount of data in the OMIM database was comparatively small.In contrast,the amount of data in the CTD database was vast,thus we selected either the top 500 targets or those with an inference score > 10.The inference score indicates a correlation between the active compound and its target,with higher values indicating stronger correlation[19].After eliminating duplicate values,601 targets of influenza viruses were retained.

    2.3 Protein-protein interaction(PPI)networks

    In order to visualize functions and interactions between target proteins,the compiled influenza targets were processed using the STRING 10.0 database[20](https://string-db.org/)to produce a PPI network of influenza target proteins.After importing the network into Cytoscape 3.4.0 software[21],it was matched with the component-target network to get the target protein which component act on influenza by.The target protein was entered into STRING 10.0 to produce a PPI network of the influenza target protein,and the network was processed using Cytoscape to examine and analyze its topology properties.

    2.4 Gene Ontology(GO)functional enrichment analysis

    The gene target was identified by processing the core target of the above analysis in the WebGestalt[22]database(http://www.webgestalt.org/)so as to elucidate genetic effects of the main components of MXSGD.

    2.5 Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis

    The KEGG pathway enrichment analysis was performed on the core target obtained by the above analysis using the reactome database[23](https://reactome.org)to understand the role of the target protein of the main component of MXSGD in the signaling pathway and to examine how MXSGD may exert beneficial effects in influenza treatments.Results of the enrichment analysis were visualized using Omicshare software(http://www.omicshare.com/tools/index.php/).

    2.6 iGEMDOCK molecular docking

    iGEMDOCK[24]is an analysis tool based on docking poses,that is,protein-ligand interactions and compound properties,and results can be visualized directly using molecular visualization tools.Docking results can be produced after uploading the protein structure files from the Protein Data Bank(http://www.rcsb.org/),and ligand files can be obtained from the ZINC(http://zinc.docking.org/)or Pub Chem(http://pubchem.ncbi.nlm.nih.gov/)online compound databases in pdb and mol2 format,respectively.The term “fitness” in the energy column refers to the total energy of the predicted pose at the binding site.The empirical scoring function of iGEMDOCK is estimated as follows:fitness=vdW + Hbond + Elec,where vdW is Van der Waal energy,and Hbond and Elect are hydrogen bond energy and electronic statistical energy,respectively.The degree of binding is judged by the level of energy,i.e.,lower energy indicates higher stability of the bond between compound and protein.The function Autodock was used to identify the docking site,and PyMOL was used to visualize docking results.

    3 Results

    3.1 Compilation and screening of MXSGD compounds and corresponding targets

    A total of 657 components of MXSGD were collected using the TCMSP database,including 363 compounds of Ephedra Herba(Ma Huang,麻黃),113 compounds of Armeniacae Semen Amarum(Ku Xing Ren,苦杏仁),280 compounds of Glycyrrhizae Radix Et Rhizoma(Gan Cao,甘草),and one compound of Gypsum Fibrosum(Shi Gao,石膏).Using screening thresholds of OB ≥ 30% and DL ≥ 0.18,135 compounds were produced,and 126 types of main compounds of MXSGD were retained after eliminating duplicate values(Table 1).After compiling the corresponding targets of the main compounds of MXSGD using the TCMSP database and the STITCH platform,we can get 260 targets of MXSGD.

    3.2 Construction of a PPI network

    Component target proteins identified from the TCMSP database were imported into the STRING 10.0 database and were examined to produce an influenza PPI network.The network was processed using Cytoscape and was merged with the compound-target network to identify influenza protein targets of MXSGD.These targets were processed using STRING 10.0 to produce a PPI network of MXSGD affecting influenza target proteins which was then topologically analyzed using Cytoscape.The PPI network of influenza(Figure 1)contained 355 influenza target proteins and 1 221 influenza protein interactions.Results of the Network Analyzer of Cytoscape suggested that the genes Tp53,MAPK1 and RELA are key targets of influenza viruses.The PPI network of influenza target proteins of MXSGD(Figure 2)contained 52 target proteins and 672 protein interactions.Results of the Network Analyzer of Cytoscape revealed the topologies of MXSGD in the PPI network of influenza target proteins.Targets are shown in Table 2,ranked by degree.The targets included Tp53,AKT1,IL-6,JUN and MAPK8,among others.

    Table 1 Active compounds of MXSGD

    Table 1 Continued

    Table 1 Continued

    Table 1 Continued

    3.3 GO functional analysis

    GO functional enrichment analysis was performed on 52 core targets involved in the PPI network using the WebGestalt database.This produced 12 biological processes including responses to stimuli,biological regulations and metabolic processes,19 cellular components including nucleus,membraneenclosed lumen and cytosol,and 14 molecular functions including protein binding,ion binding and transferase activity(Figure 3).

    3.4 KEGG pathway enrichment analysis

    KEGG pathway enrichment analysis were performed on the core target protein of the PPI network of agglutinin in the influenza using the reactome database.Results were screened using aP<0.01 and with an FDR<0.05.The top 20 samples were visualized using the Omicshare platform(Figure 4).The color band on the right indicates the main enrichment pathway,withPvaluesincreasing from red to purple.We suggest that the effects of MXSGD on influenza involve multiple complex biological processes.MXSGD acts on targets at the core of the influenza PPI network.The predominantly involved pathways were cytokine signaling in the immune system(Figure 5)and interleukin signaling(Figure 6),among others(Table 3).

    3.5 Results of iGEMDOCK molecular docking

    Table 2 Topological properties of MXSGD compounds acting on influenza target proteins

    From the above results,we conclude that the top three core targets during influenza treatment are Tp53,MAPK1 and RELA.These three core targets are also MXSGD targets during influenza treatment,and the top three core targets of MXSGD during influenza treatment were Tp53,AKT1 and IL-6.The five targets Tp53,MAPK1,RELA,AKT1 and IL-6 were selected for docking and comparison with the corresponding active compounds and with oseltamivir and amantadine,which are commonly used in clinical research for treating influenza(Figure 7).The docking results show that,compared with oseltamivir and amantadine,MXSGD compounds produced lower binding energy and were more stably bound to their respective receptors.The lowest binding energy was observed in RELA-Licochalcone A,followed by RELA-Quercetin and RELA-Naringenin.The specific docking site is shown in Figure 8.The binding site of RELA and Licochalcone A is amino acid 441 of the RELA protein,the binding site of RELA and Quercetin is constituted by amino acids 64 and 80 of the RELA protein,and the binding site of RELA and Naringenin is constituted by amino acids 322 and 325 of the RELA protein.

    Table 2 Continued

    4 Discussion

    Network pharmacology integrates the three aspects target,drug and disease,using them as intermediate nodes,and the links between the three aspects are regarded as edges,thereby producing specific information through complex network analysis methods[25-29].This approach may be used to visualize the multi-compound,multi-target and multi-path effects of TCM through network construction.MXSGD was first mentioned inTreatise on Cold Damage Diseases(Shang Han Lun,《傷寒論》),and it is predominantly used to treat symptoms such as cough,asthma and even nasal ale flaps or high fever[30].Recent research showed that MXSGD can be used to treat a variety of lung diseases[31-36].The current study is based on network pharmacology and analyzes network characteristics through the connection and relationship of nodes in biological networks in order to explore the mechanism of MXSGD in influenza treatments.

    Table 3 KEGG pathway enrichment during MXSGD-based influenza treatment

    According to the above results,treatment of influenza using MXSGD involves 52 main targets,12 biological processes,19 cellular components,14 molecular functions and 20 major KEGG pathways.We analyzed topological properties of 52 important targets and assessed closeness centrality and average length of the shortest path from each node to other nodes in the topology of network nodes.The degree indicates the number of neighboring nodes of any one node[37].The advantage of this indicator value is that it is intuitive and simple to calculate,however,it does not necessarily reflect the significance of any one node in the network accurately.“Betweenness centrality” is based on complex calculations to characterize the role of the node within the network,to indicate the number of shortest paths between all pairs of connected nodes,and to estimate the load of a node within the network[38].After comprehensive analysis of topological properties,Tp53,AKT1,IL-6,JUN,MAPK8 and MAPK1,among others,were identified as important targets.Studies have shown that microRNA(miRNA)commonly occurs in viruses and eukaryotes and participates in the regulation of almost all cell physiological processes that have been studied in the past[39-44].Abnormal miRNA expression has been observed during infection with influenza,and miRNAs are strongly correlated with regulation of AKT1,JUN and other genes.MXSGD was suggested to interact with genes such as AKTI and JUN to regulate miRNA and thereby inhibit influenza viruses.As shown in Figure 6,interleukin signaling pathways including those of IL-2,IL-4,IL-6 and IL-10 can affect interleukin signaling through the STAT gene[45,46],and previous studies showed that STAT1 can control early-stage replication of influenza viruses[47].The interleukin family plays an important role in cytokine signaling pathways of the immune system,suggesting that interleukins are involved in regulating the immune system,and MXSGD can interfere with excessive immune responses by intervening interleukins.The Tp53 gene was found to exert antiviral effects by inhibiting virus replication and increasing innate and adaptive immune competence through IFN[48].Thus,MXSGD may help counteract influenza infections by regulating Tp53 expression so as to inhibit virus replication and improve the respective immune functions.Taken together,we conclude that MXSGD may exert anti-influenza virus effects,predominantly by regulating RNA expression to inhibit virus replication and overexpression of inflammation factors,and regulate the immune system.

    In the KEGG enrichment pathway,MXSGD appeared to affect RNA polymerase by reducing its positive regulatory effect and thus inhibiting virus replication.Influenza viruses may also be inhibited by regulating genes such as AKTI and JUN,among others,to mediate general transcription pathways,RNA polymerase II transcription pathways,gene expression(transcription)pathways,and regulation of ribozyme and nucleotides in biological processes.In addition,the GO functional and KEGG pathway analyses indicated that MXSGD affected the genes IL-2,IL-4,IL-6 and Tp53,among others.These factors can promote immune cell proliferation which can cause excessive inflammatory reactions.MXSGD can thus reduce excessive inflammatory processes by inhibiting gene expression.In a study on lipid and protein metabolism,MXSGD was found to be associated with inhibition of neuraminidase during influenza treatment[49],which subsequently induced production of specific humoral and cellular immune factors.Taken together,MXSGD exerts anti-influenza effects by interfering with virus absorption,inhibiting virus proliferation,regulating the immune system and protecting host cells.

    Molecular docking results in this study indicated that the bond of the target protein rela and the drug was most stable.Combined with Licochalcone A,Quercetin and Naringenin from Ephedra Herba(Ma Huang,麻黃)and Glycyrrhizae Radix Et Rhizoma(Gan Cao,甘草),the binding energy is reduced which should increase its efficiency.Although most compounds of MXSGD stemmed from Glycyrrhizae Radix Et Rhizoma(Gan Cao,甘草),the main compounds with more stable binding to the target originated from Ephedra Herba(Ma Huang,麻黃),which is a sovereign medicine.Previous studies have shown that both licochalcone a and naringenin can inhibit activation of NF-κB and exert anti-inflammatory effects[50,51].NF-κB is a key protein in immune and inflammatory responses,and RELA is a subunit of NFκB[52].Quercetin was found to possess antiviral,antiinflammatory,anti-oxidant and vasodilatory effects[53]; therefore,Licochalcone A,Quercetin and Naringenin of Ephedra Herba(Ma Huang,麻黃)and Glycyrrhizae Radix Et Rhizoma(Gan Cao,甘草)may have certain anti-inflammatory effects by inhibiting the expression of RELA and by preventing excessive inflammation and inflammation-induced tissue damage.

    In the current study,we identified compounds of MXSGD that are potentially active against influenza using systematic pharmacology,and we used a molecular docking method to comprehensively elucidate therapeutic mechanisms of MXSGD in influenza treatments.Our results are mostly consistent with those of previous studies.Although our results are very interesting,further research and experimental data would be needed to clarify mechanisms such as drug dosage effects,whether miRNAs are mediated by the AKT1 gene to inhibit influenza viruses,and whether two-way regulations of interleukins may occur.In addition,the molecular mechanisms of active compounds should be testedin vivoto evaluate potential clinical application.

    Acknowledgements

    We thank for the funding support from the National Natural Science Foundation of China(No.81973670),the Natural Science Foundation of Hunan Province(No.2018JJ2297),the Key Program of Scientific Research Fund of Hunan Provincial Education Department(No.19A370)and the Project of Research Learning and Innovative Experiment for College Students in Hunan(No.2016284,No.2016281,No.2017281and No.2018420).

    Competing Interests

    The authors declare no conflict of interest.

    猜你喜歡
    傷寒論麻黃石膏
    General principle of high-quality academic development of traditional chinese medicine: “carrying on the essence,while pursuing innovations”
    《傷寒論》六經(jīng)溯源與理論淺析
    國(guó)內(nèi)外磷石膏綜合利用現(xiàn)狀
    云南化工(2021年11期)2022-01-12 06:06:06
    孔伯華與石膏
    《傷寒論》實(shí)為115方
    A Patient with End-stage Respiratory Failure Gets Recovery from Mechanical Ventilation by Utilization of Chinese Traditional Medicine
    Systematic Pharmacological Strategies to Explore the Regulatory Mechanism of Ma Xing Shi Gan Decoction on COVID-19
    關(guān)注麻黃的配伍禁忌
    石膏之魂
    西部論叢(2019年17期)2019-10-14 11:09:40
    麻黃-杏仁藥對(duì)成分-效應(yīng)間變化規(guī)律
    中成藥(2018年6期)2018-07-11 03:01:18
    av中文乱码字幕在线| 美女 人体艺术 gogo| 国产精品久久久久久久久免 | 伊人久久精品亚洲午夜| ponron亚洲| 欧美黑人欧美精品刺激| 在线观看午夜福利视频| 亚洲一区高清亚洲精品| 人人妻人人看人人澡| 精品一区二区三区人妻视频| 少妇的逼好多水| 性插视频无遮挡在线免费观看| 久久久国产成人精品二区| 99精品在免费线老司机午夜| 我的老师免费观看完整版| 国产精品一及| 中国美女看黄片| 久久精品国产99精品国产亚洲性色| 国产欧美日韩精品一区二区| 日韩精品青青久久久久久| 亚洲内射少妇av| 午夜精品在线福利| 国产在视频线在精品| 欧美日韩瑟瑟在线播放| 一级a爱片免费观看的视频| 国产精品自产拍在线观看55亚洲| 色视频www国产| 免费av观看视频| 99久久久亚洲精品蜜臀av| 久久99热这里只有精品18| 亚洲欧美日韩卡通动漫| 如何舔出高潮| 国产黄片美女视频| 少妇丰满av| 最近中文字幕高清免费大全6 | 免费黄网站久久成人精品 | 男人的好看免费观看在线视频| 久久伊人香网站| 日韩精品中文字幕看吧| 精华霜和精华液先用哪个| 日韩欧美免费精品| 国产成人av教育| 麻豆av噜噜一区二区三区| 亚洲精品乱码久久久v下载方式| 日本撒尿小便嘘嘘汇集6| 99国产综合亚洲精品| 精品久久久久久久末码| 国内精品久久久久久久电影| 欧美高清性xxxxhd video| 在线播放国产精品三级| 小蜜桃在线观看免费完整版高清| 国产精品不卡视频一区二区 | 欧美日韩中文字幕国产精品一区二区三区| 99久久无色码亚洲精品果冻| 一级黄片播放器| 免费黄网站久久成人精品 | 中文字幕熟女人妻在线| 给我免费播放毛片高清在线观看| 精品日产1卡2卡| 麻豆一二三区av精品| 精品不卡国产一区二区三区| 成人国产一区最新在线观看| 三级国产精品欧美在线观看| 搡老岳熟女国产| 婷婷六月久久综合丁香| 少妇的逼好多水| 最近视频中文字幕2019在线8| 久久精品影院6| 亚洲av一区综合| 深夜精品福利| 亚洲无线在线观看| 国产一区二区三区在线臀色熟女| 欧美一区二区亚洲| 亚洲国产欧美人成| 国产高清有码在线观看视频| 国模一区二区三区四区视频| 青草久久国产| 一级黄片播放器| 校园春色视频在线观看| 欧美极品一区二区三区四区| 99久久成人亚洲精品观看| 日韩大尺度精品在线看网址| 国产在线男女| 亚洲国产精品合色在线| 两个人的视频大全免费| 五月伊人婷婷丁香| 极品教师在线免费播放| 国产精品伦人一区二区| 99久久九九国产精品国产免费| 久久精品夜夜夜夜夜久久蜜豆| 夜夜爽天天搞| 中文字幕久久专区| 欧美一区二区精品小视频在线| 午夜福利免费观看在线| 首页视频小说图片口味搜索| 国产黄片美女视频| 亚洲av美国av| aaaaa片日本免费| 亚洲无线观看免费| 美女免费视频网站| 一个人观看的视频www高清免费观看| 国内久久婷婷六月综合欲色啪| .国产精品久久| 免费电影在线观看免费观看| 国产成人av教育| 搡老熟女国产l中国老女人| 亚洲成人精品中文字幕电影| 免费人成视频x8x8入口观看| 天堂网av新在线| 亚洲专区中文字幕在线| 99国产综合亚洲精品| 婷婷丁香在线五月| 国产黄片美女视频| 欧美黑人欧美精品刺激| 无人区码免费观看不卡| netflix在线观看网站| 久久国产乱子免费精品| 天天躁日日操中文字幕| 真实男女啪啪啪动态图| 国产爱豆传媒在线观看| 欧美性感艳星| 国产 一区 欧美 日韩| av在线蜜桃| 精品人妻熟女av久视频| 免费在线观看日本一区| 我的老师免费观看完整版| 床上黄色一级片| 亚洲欧美激情综合另类| 久久久久国产精品人妻aⅴ院| 九九久久精品国产亚洲av麻豆| 国产精华一区二区三区| 久久伊人香网站| 观看美女的网站| 午夜精品久久久久久毛片777| 亚洲精品在线观看二区| 国产免费男女视频| 国产aⅴ精品一区二区三区波| 国产亚洲精品久久久com| 欧美黄色淫秽网站| 久久精品国产亚洲av涩爱 | 久久精品91蜜桃| 亚洲男人的天堂狠狠| 一a级毛片在线观看| 一个人看的www免费观看视频| 日本在线视频免费播放| 欧美色视频一区免费| 色精品久久人妻99蜜桃| 午夜亚洲福利在线播放| 天美传媒精品一区二区| 欧美日韩中文字幕国产精品一区二区三区| 久久亚洲精品不卡| 亚洲乱码一区二区免费版| 亚洲国产精品sss在线观看| 免费看a级黄色片| 国产精品人妻久久久久久| 久久久色成人| 亚洲欧美清纯卡通| 精品国内亚洲2022精品成人| 日韩免费av在线播放| 淫秽高清视频在线观看| 亚洲第一欧美日韩一区二区三区| 成人特级黄色片久久久久久久| 日本精品一区二区三区蜜桃| 国产亚洲欧美98| 在线十欧美十亚洲十日本专区| 最新中文字幕久久久久| 美女cb高潮喷水在线观看| 99热这里只有是精品50| 嫩草影视91久久| 美女xxoo啪啪120秒动态图 | 村上凉子中文字幕在线| 日韩av在线大香蕉| 天堂影院成人在线观看| 午夜福利高清视频| 国内毛片毛片毛片毛片毛片| 乱码一卡2卡4卡精品| 国产极品精品免费视频能看的| 国产av不卡久久| 日韩有码中文字幕| 久久久久久久久久成人| 久久99热这里只有精品18| 毛片女人毛片| 特级一级黄色大片| 精品国产三级普通话版| 1024手机看黄色片| 久久久久久久久久成人| 一进一出好大好爽视频| 熟女人妻精品中文字幕| 天堂网av新在线| 国产av麻豆久久久久久久| 国语自产精品视频在线第100页| 99热只有精品国产| 国产中年淑女户外野战色| 欧美绝顶高潮抽搐喷水| 欧美午夜高清在线| 熟女人妻精品中文字幕| 国产毛片a区久久久久| 国产91精品成人一区二区三区| 别揉我奶头~嗯~啊~动态视频| 欧美高清成人免费视频www| 男女下面进入的视频免费午夜| 嫁个100分男人电影在线观看| 亚洲在线观看片| 神马国产精品三级电影在线观看| 中国美女看黄片| 国产欧美日韩精品一区二区| 国产伦精品一区二区三区视频9| 亚洲最大成人手机在线| 国产亚洲精品av在线| 国产主播在线观看一区二区| 国产成人av教育| 特级一级黄色大片| 97超视频在线观看视频| aaaaa片日本免费| 国语自产精品视频在线第100页| 有码 亚洲区| 欧美3d第一页| 黄色丝袜av网址大全| 免费人成在线观看视频色| 2021天堂中文幕一二区在线观| 国模一区二区三区四区视频| 国产伦一二天堂av在线观看| 一二三四社区在线视频社区8| 成年免费大片在线观看| 看十八女毛片水多多多| 90打野战视频偷拍视频| 尤物成人国产欧美一区二区三区| 亚洲欧美日韩无卡精品| 高清毛片免费观看视频网站| 757午夜福利合集在线观看| 精品午夜福利在线看| 桃色一区二区三区在线观看| 精品一区二区三区人妻视频| 日韩欧美 国产精品| 亚洲av第一区精品v没综合| 国产又黄又爽又无遮挡在线| 亚洲精品成人久久久久久| 真人一进一出gif抽搐免费| 在线十欧美十亚洲十日本专区| 人人妻,人人澡人人爽秒播| 欧美日本亚洲视频在线播放| 99久久精品热视频| 精品免费久久久久久久清纯| 麻豆成人午夜福利视频| 国产久久久一区二区三区| 国产伦一二天堂av在线观看| av在线蜜桃| 国产精品,欧美在线| 俺也久久电影网| 韩国av一区二区三区四区| 亚洲,欧美精品.| 欧美黄色淫秽网站| 91久久精品电影网| 国产伦人伦偷精品视频| 成人一区二区视频在线观看| 亚洲,欧美精品.| 国产精品久久久久久久久免 | 毛片一级片免费看久久久久 | 日韩国内少妇激情av| 哪里可以看免费的av片| 欧美高清性xxxxhd video| 在线观看av片永久免费下载| 日本一二三区视频观看| 伊人久久精品亚洲午夜| 琪琪午夜伦伦电影理论片6080| 亚洲精品乱码久久久v下载方式| 午夜福利高清视频| 欧美黄色片欧美黄色片| 亚洲av一区综合| 亚洲精品色激情综合| 嫩草影院精品99| 免费人成视频x8x8入口观看| 毛片一级片免费看久久久久 | 国产亚洲欧美在线一区二区| 我要搜黄色片| 亚洲欧美日韩高清专用| 日本三级黄在线观看| 国产午夜精品论理片| 精品一区二区三区视频在线观看免费| 搡女人真爽免费视频火全软件 | 欧美日韩瑟瑟在线播放| 久久精品国产亚洲av香蕉五月| aaaaa片日本免费| 亚洲一区二区三区色噜噜| 99久久无色码亚洲精品果冻| 亚洲一区二区三区不卡视频| 国产精品日韩av在线免费观看| 九色国产91popny在线| 国产一区二区在线观看日韩| 欧美3d第一页| 色在线成人网| 免费在线观看成人毛片| 日韩人妻高清精品专区| 久久午夜亚洲精品久久| 免费av不卡在线播放| 一夜夜www| netflix在线观看网站| 国产欧美日韩精品亚洲av| 高清在线国产一区| 欧美在线黄色| 婷婷丁香在线五月| 人人妻人人澡欧美一区二区| 日韩有码中文字幕| 人妻夜夜爽99麻豆av| 午夜老司机福利剧场| 日韩国内少妇激情av| 听说在线观看完整版免费高清| 免费人成在线观看视频色| 在线免费观看的www视频| 99热这里只有精品一区| 无遮挡黄片免费观看| 狂野欧美白嫩少妇大欣赏| 欧美日韩黄片免| 精品久久久久久久末码| 国产乱人伦免费视频| 在线观看美女被高潮喷水网站 | 亚洲,欧美精品.| 久久精品国产亚洲av天美| 亚洲成人中文字幕在线播放| 精品午夜福利视频在线观看一区| 久久久久亚洲av毛片大全| 最近在线观看免费完整版| 精品不卡国产一区二区三区| 97热精品久久久久久| 成年版毛片免费区| 日日干狠狠操夜夜爽| 韩国av一区二区三区四区| 久久久久久久午夜电影| 免费人成在线观看视频色| 精品午夜福利在线看| 最近最新中文字幕大全电影3| 午夜a级毛片| 老司机午夜福利在线观看视频| 99国产极品粉嫩在线观看| 狂野欧美白嫩少妇大欣赏| 国产高清视频在线播放一区| 日韩亚洲欧美综合| 亚洲第一区二区三区不卡| 99riav亚洲国产免费| 在线观看66精品国产| 99国产精品一区二区蜜桃av| 黄色配什么色好看| 两个人视频免费观看高清| 内射极品少妇av片p| 亚洲狠狠婷婷综合久久图片| 国产精品人妻久久久久久| 欧美一区二区国产精品久久精品| 亚洲av中文字字幕乱码综合| 亚洲最大成人av| 精品久久久久久久人妻蜜臀av| 国产国拍精品亚洲av在线观看| 国产色爽女视频免费观看| 国产精品久久久久久精品电影| 一卡2卡三卡四卡精品乱码亚洲| 五月伊人婷婷丁香| 精品午夜福利视频在线观看一区| av欧美777| 亚洲avbb在线观看| 午夜a级毛片| 精品久久久久久成人av| 精品国产三级普通话版| 日本三级黄在线观看| 国产精品av视频在线免费观看| 色综合欧美亚洲国产小说| 欧美在线黄色| 内地一区二区视频在线| 国产主播在线观看一区二区| 亚洲天堂国产精品一区在线| 在线免费观看的www视频| 又黄又爽又刺激的免费视频.| 亚洲激情在线av| 国产综合懂色| 五月玫瑰六月丁香| 99精品在免费线老司机午夜| 露出奶头的视频| 午夜日韩欧美国产| 国产精品自产拍在线观看55亚洲| 一个人看视频在线观看www免费| 男女之事视频高清在线观看| 免费无遮挡裸体视频| 欧美黑人巨大hd| 国产白丝娇喘喷水9色精品| 99国产精品一区二区蜜桃av| 麻豆成人av在线观看| 欧美xxxx黑人xx丫x性爽| 在线观看66精品国产| 97热精品久久久久久| 熟女电影av网| 精品欧美国产一区二区三| 欧美日韩黄片免| 国产淫片久久久久久久久 | 欧美xxxx黑人xx丫x性爽| 麻豆成人av在线观看| 俺也久久电影网| 永久网站在线| 亚洲av成人av| 可以在线观看毛片的网站| 国产黄a三级三级三级人| 国产欧美日韩精品一区二区| 国产精品精品国产色婷婷| 亚洲无线观看免费| 欧美zozozo另类| 亚洲精品一卡2卡三卡4卡5卡| 精品久久久久久久末码| 午夜福利在线观看免费完整高清在 | 非洲黑人性xxxx精品又粗又长| 大型黄色视频在线免费观看| 欧美黄色淫秽网站| 一区福利在线观看| 午夜久久久久精精品| 国产精品,欧美在线| 午夜精品一区二区三区免费看| 床上黄色一级片| 在线国产一区二区在线| 免费观看人在逋| 国产毛片a区久久久久| 亚洲五月天丁香| 日本黄色视频三级网站网址| 午夜精品久久久久久毛片777| 嫩草影视91久久| 成年免费大片在线观看| 午夜a级毛片| 午夜久久久久精精品| www.999成人在线观看| 免费观看精品视频网站| 亚洲内射少妇av| 中文字幕av在线有码专区| 国产精品自产拍在线观看55亚洲| 欧美激情久久久久久爽电影| 一边摸一边抽搐一进一小说| 一进一出抽搐gif免费好疼| 欧美xxxx性猛交bbbb| 亚洲精品一区av在线观看| 男人狂女人下面高潮的视频| 久久精品国产亚洲av香蕉五月| 国产高清三级在线| 久久久久久大精品| 成人高潮视频无遮挡免费网站| 国产精品久久视频播放| 欧美日韩黄片免| 在线免费观看不下载黄p国产 | 欧美zozozo另类| 国产av一区在线观看免费| av中文乱码字幕在线| 国产精品美女特级片免费视频播放器| 亚洲va日本ⅴa欧美va伊人久久| 欧美日韩中文字幕国产精品一区二区三区| 亚洲av成人精品一区久久| 国产精品久久久久久精品电影| 中出人妻视频一区二区| av天堂在线播放| 狂野欧美白嫩少妇大欣赏| 12—13女人毛片做爰片一| 久久久久九九精品影院| 国产探花在线观看一区二区| 欧美日韩瑟瑟在线播放| 亚洲av.av天堂| 简卡轻食公司| 亚洲精品在线美女| 久久久久九九精品影院| 成人永久免费在线观看视频| 国产精品日韩av在线免费观看| 亚洲国产精品久久男人天堂| 精品午夜福利视频在线观看一区| 老司机深夜福利视频在线观看| 午夜福利免费观看在线| 国产在线精品亚洲第一网站| 热99在线观看视频| 99热6这里只有精品| 身体一侧抽搐| 午夜免费激情av| 一个人看的www免费观看视频| av视频在线观看入口| 两性午夜刺激爽爽歪歪视频在线观看| 亚洲性夜色夜夜综合| 国产伦精品一区二区三区四那| 国产成年人精品一区二区| 一本精品99久久精品77| 丰满的人妻完整版| 男人狂女人下面高潮的视频| 色在线成人网| 一级av片app| 午夜影院日韩av| 国产亚洲欧美98| 村上凉子中文字幕在线| 色视频www国产| 国产三级在线视频| 夜夜夜夜夜久久久久| 国产精品久久久久久亚洲av鲁大| 简卡轻食公司| 欧美一区二区精品小视频在线| 国产人妻一区二区三区在| 午夜福利高清视频| 简卡轻食公司| 亚洲乱码一区二区免费版| 黄色丝袜av网址大全| 国产亚洲欧美在线一区二区| 99久久九九国产精品国产免费| 亚洲最大成人手机在线| 村上凉子中文字幕在线| av在线老鸭窝| 免费大片18禁| 午夜两性在线视频| 桃色一区二区三区在线观看| 我的女老师完整版在线观看| 亚洲午夜理论影院| 色精品久久人妻99蜜桃| 此物有八面人人有两片| 在线播放国产精品三级| 丰满乱子伦码专区| 99久久九九国产精品国产免费| 精品一区二区三区视频在线| 3wmmmm亚洲av在线观看| 精品一区二区三区av网在线观看| 欧美性感艳星| 中文亚洲av片在线观看爽| 非洲黑人性xxxx精品又粗又长| 亚洲美女视频黄频| 日日摸夜夜添夜夜添小说| 丝袜美腿在线中文| 日韩人妻高清精品专区| 内地一区二区视频在线| 欧美日韩中文字幕国产精品一区二区三区| h日本视频在线播放| 一进一出抽搐动态| 久久亚洲精品不卡| 少妇的逼水好多| 免费人成视频x8x8入口观看| 在线观看av片永久免费下载| 亚洲国产精品久久男人天堂| 90打野战视频偷拍视频| 欧美日韩综合久久久久久 | 成人av在线播放网站| 亚洲精品日韩av片在线观看| 啦啦啦观看免费观看视频高清| 国内毛片毛片毛片毛片毛片| 成年女人看的毛片在线观看| 性欧美人与动物交配| 在线免费观看的www视频| 国产成+人综合+亚洲专区| 国产精华一区二区三区| 村上凉子中文字幕在线| 久久久精品大字幕| 观看美女的网站| 国内精品美女久久久久久| 精品午夜福利视频在线观看一区| 变态另类丝袜制服| 婷婷精品国产亚洲av| 亚洲人成伊人成综合网2020| 一进一出好大好爽视频| 欧美日韩国产亚洲二区| 午夜精品在线福利| 久久国产乱子免费精品| 99精品在免费线老司机午夜| 欧美成人a在线观看| 久久欧美精品欧美久久欧美| 国产综合懂色| 男女之事视频高清在线观看| 一区二区三区高清视频在线| 欧美极品一区二区三区四区| 欧美日韩中文字幕国产精品一区二区三区| 免费av不卡在线播放| xxxwww97欧美| 美女高潮喷水抽搐中文字幕| 少妇熟女aⅴ在线视频| 最近在线观看免费完整版| 一区福利在线观看| netflix在线观看网站| 免费观看的影片在线观看| 亚洲精品成人久久久久久| 国产精品野战在线观看| 亚洲五月天丁香| 最好的美女福利视频网| 精品久久久久久久久久免费视频| а√天堂www在线а√下载| 每晚都被弄得嗷嗷叫到高潮| 亚洲avbb在线观看| 我要看日韩黄色一级片| 一区二区三区激情视频| 国产单亲对白刺激| 有码 亚洲区| 精品不卡国产一区二区三区| 男人狂女人下面高潮的视频| 国产一区二区在线av高清观看| 国产乱人伦免费视频| 亚洲成a人片在线一区二区| 中亚洲国语对白在线视频| 日本五十路高清| 天堂网av新在线| 久久亚洲真实| 免费看a级黄色片| 中文字幕久久专区| 久久天躁狠狠躁夜夜2o2o| 无遮挡黄片免费观看| 美女被艹到高潮喷水动态| 在线观看午夜福利视频| 村上凉子中文字幕在线| 可以在线观看毛片的网站| 在线观看午夜福利视频| 亚洲成人中文字幕在线播放| 精品一区二区三区av网在线观看| 国产精品久久久久久人妻精品电影| 91午夜精品亚洲一区二区三区 | 国产一区二区三区视频了| 色av中文字幕| 乱人视频在线观看| 悠悠久久av| 午夜久久久久精精品| av视频在线观看入口| 午夜免费成人在线视频| 国产 一区 欧美 日韩| 91在线精品国自产拍蜜月| 国产精品久久久久久亚洲av鲁大| 天堂网av新在线| 一区二区三区免费毛片|