Four new 3,4-seco-labdane diterpenoids,nudiflopenes J-M,were isolated from the leaves of Callicarpa nudiflora along with six known compounds.The structures of these diterpenoids were determined by comprehensive spectr...Four new 3,4-seco-labdane diterpenoids,nudiflopenes J-M,were isolated from the leaves of Callicarpa nudiflora along with six known compounds.The structures of these diterpenoids were determined by comprehensive spectroscopic analysis.All the isolated compounds were evaluated for their inhibitory effects on NO production in LPS-stimulated RPMs and RAW264.7 cells.The results suggest that nudiflopenes J-M and other four known compounds showed significant inhibitory effects against NO production comparable to the positive control dexamethasone.展开更多
Objective:To evaluate the antidiabetic effect of Callicarpa nudiflora extract in streptozotoc-ininduced diabetic rats.Methods:The chemical constituents in Callicarpa nudiflora extract were identified by UPLCQ-TOF-MS.C...Objective:To evaluate the antidiabetic effect of Callicarpa nudiflora extract in streptozotoc-ininduced diabetic rats.Methods:The chemical constituents in Callicarpa nudiflora extract were identified by UPLCQ-TOF-MS.Callicarpa nudiflora extract(0.15 and 0.3 g/kg)was orally administered to streptozotocin-induced diabetic rats for 42 d.The effects of Callicarpa nudiflora extract on body weight,blood glucose,serum insulin,total cholesterol,triglyceride,LDL-C and HDL-C were investigated,and its effect on liver and pancreatic pathology was assessed by histopathological analysis.Moreover,the expression levels of adenosine 5’-monophosphate-activated protein kinase(AMPK),glucose transporter type 4(GLUT4),phospho-AMPK/AMPK,and p-acetylcoA carboxylase(P-ACC)/ACC in the skeletal muscles and liver were determined by reverse transcription-polymerase chain reaction,Western blotting,and immunohistochemistry.Results:A total of 34 compounds,including 8 iridoids,14 phenylpropanoids,and 12 flavonoids,were identified from Callicarpa nudiflora extract.Callicarpa nudiflora extract significantly reduced blood glucose and significantly restored all other biochemical parameters to near normal levels,including serum insulin,total cholesterol,triglyceride,LDL-C,and HDL-C.Callicarpa nudiflora extract improved insulin resistance and reversed the damage in the liver and pancreas caused by diabetes.Furthermore,Callicarpa nudiflora extract increased the expression levels of phospho-AMPK,GLUT4,P-ACC,and insulin receptor substrate-1 and decreased the expression level of PPAR r in diabetic rats.Conclusions:Callicarpa nudiflora extract improved oral glucose tolerance,lipid metabolism,insulin resistance,and reversed the diabetes-related damage in the liver and pancreas by activating the AMPK-ACC pathway.展开更多
A phytochemical investigation was carried out on the extract of a medicinal plant Callicarpa nudiflora,resulting in the characterization of five new 3,4-seco-isopimarane(1-5)and one new 3,4-seco-pimarane diterpenoid(6...A phytochemical investigation was carried out on the extract of a medicinal plant Callicarpa nudiflora,resulting in the characterization of five new 3,4-seco-isopimarane(1-5)and one new 3,4-seco-pimarane diterpenoid(6),together with four known compounds.The structures of the new compounds were fully elucidated by extensive analysis of MS,1 D and 2 D NMR spectroscopic data,and time-dependent density functional theory(TDDFT)calculation of electronic circular dichroism(ECD)spectra,and DFT calculations for NMR chemical shifts and optical rotations.展开更多
[Objective] This study aimed to study the coagulation effect of Callicarpa nudiflora. [Method] The effects of CaUtcarpa nudiflora on blood coagulation system of mice were investigated by measuring prothmmbin time （PT...[Objective] This study aimed to study the coagulation effect of Callicarpa nudiflora. [Method] The effects of CaUtcarpa nudiflora on blood coagulation system of mice were investigated by measuring prothmmbin time （PT）, thrombin time（TT）, activated partial thromboplastin time （APTT） and plasma fibrinogen （FIB）, [Result] Compared with the control group, the crude extracts and ：eluted parts of Ca#/carpa nudiflora all significantly shortened PT, APTT and TT, and increased the content of FIB in mice, [Coadadon] The crude extracts and eluted parts of Callicarpa nudiflora have a coagulation-promoting effect through affecting several links of the coagulation process in mice,展开更多
Objective: To study the effect of the extracts from Callicarpa nudiflora on wound healing and the preparation of ointment. Methods: Totally 50 KM mice were randomly divided into five groups, which were negative contro...Objective: To study the effect of the extracts from Callicarpa nudiflora on wound healing and the preparation of ointment. Methods: Totally 50 KM mice were randomly divided into five groups, which were negative control group ,positive control group, high, middle and low dose groups of Callicarpa nudiflora ointment. Mice ear swelling test was conducted to investigate the effect of Callicarpa nudiflora induced by xylene, Bacteriostasis experiments were conducted to observe the bacteriostasis effects of Callicarpa nudiflora on five indicator bacteria, i.e. Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Bacillus subtilis, and Staphylococcus aureus in vitro. A total of 30 new zealand white rabbit with half male and half female were randomly divided into five groups, which were negative control group, positive control group, high, middle and low groups of Callicarpa nudiflora ointment. The rabbits were treated with sterilizing blade and treated with external medicine to observe the effect of the extracts from Callicarpa nudiflora on wound healing. Orthogonal test was used to determine the preparation method of Callicarpa nudiflora ointment,Physical properties, centrifugal stratification test, heat and cold resistance test were conducted. Results:Bacteriostasis experiments in vitro showed high dose group of Callicarpa nudiflora extracts got 16.8 cm antimicrobial zone of Pseudomonas aeruginosa. The experimental results showed that the ear swelling degree of mice induced by xylene was significantly reduced in the middle and high dose groups. All the treatment groups could promote skin healing of rabbit significantly compared with negative control group. There were significantly differences between all the treatment groups and negative control group on the wound healing rate. The optimum preparation process was determined by orthogonal screening test. The optimal formula was as follows: Twain-801.2 g, glycerin monostearate 4.7 g, glycerinum 0.9 g, liquid paraffin 10.8 g, Span801.4 g,solid paraffin 1.4 g,vaseline 0.9 g. The emulsification temperature was 65 ℃,and emulsification time was 25 min. Conclusions: The extract of Callicarpa nudiflora can be used as external preparation and the preparation is in compliance with Chinese pharmacopoeia 2015.展开更多
Four new neo-lignans were isolated from the chloroform portion of 70% ethanol extract of the seed crusts of Trewia nudiflora L., and were determined to be 9'-methoxy-7'-en-3',8:4',7-di-epoxyneolignan-3...Four new neo-lignans were isolated from the chloroform portion of 70% ethanol extract of the seed crusts of Trewia nudiflora L., and were determined to be 9'-methoxy-7'-en-3',8:4',7-di-epoxyneolignan-3,4,9-triol l, 9'-methoxy-7'-en-3',7:4',8-diepoxyneolignan-3,4,9-triol 2, 9'- ethoxy-7'-en-3',8:4',7-diepoxyneolignan-3,4,9-triol 3, 9'-butoxy-7'-en-3',8:4',7-diepoxy-neolignan -3,4,9-triol 4, respectively, on the basis of spectral data. The antimicrobial bioassays showed that 4 had evident inhibitory activity against Mycobacterium tuberculosis.展开更多
Objective:To establish the quality standard of Li folk medicine callicarpa nudiflora powder by modern analytical method.Methods:TLC was used to identify the powder.Five heavy metals and harmful elements(Pb,Cd,As,Hg an...Objective:To establish the quality standard of Li folk medicine callicarpa nudiflora powder by modern analytical method.Methods:TLC was used to identify the powder.Five heavy metals and harmful elements(Pb,Cd,As,Hg and Cu)in the powder were determined by flame atomic absorption spectrometry and hydride atomic fluorescence spectrometry.The content of luteolin and Mullein glycoside in the powder was determined by HPLC.Results:the fluorescent spots of TLC identification of powder were clear,and the separation of the two main components was good.It has good specificity,reproducibility and durability,which can be used for the identification of the powder.The inspection items of heavy metals for the identification of powder were established.It was stipulated that:according to the determination method,the lead should not exceed 5 mg/kg;the cadmium should not exceed 0.3 mg/kg;the arsenic should not exceed 2 mg/kg;the mercury should not exceed 0.2 mg/kg;the copper should not exceed 20 mg/kg.The content of luteolin and Mullein glycoside in the powder were determined by HPLC and the content inspection item of the powder was established.It was stipulated that the content of luteolin and Mullein glycoside in the powder should not be less than 0.48%and 5.83%,which can be used for the content limit inspection of the powder.Conclusion:the quality standard method established by our institute is advanced and operable,and can be used for the quality control of the powder.展开更多
Objective:To screen the best prescription and technology of Callicarpa nudiflora film,and to study and establish the quality standard of the flim.Methods:The orthogonal test was used to investigate the effect of PVA c...Objective:To screen the best prescription and technology of Callicarpa nudiflora film,and to study and establish the quality standard of the flim.Methods:The orthogonal test was used to investigate the effect of PVA content,glycerin content,ethanol and water ratio on the comprehensive score of the coating agent;the general inspection method and content determination method were used to control the quality of Callicarpa nudiflora film,and the quality standard of the film was established.Results:The best formulation was selected as polyvinyl alcohol 6.0g,glycerin 2ml,ethanol:water 1:1.The appearance,stability,heat and cold resistance of the film were good,and the quality standard of the film was established.Conclusion:The formulation is simple and feasible,and the quality control method of the film is stable and feasible.展开更多
目的基于文献计量学方法,分析裸花紫珠的研究现状、热点及发展趋势。方法以裸花紫珠为关键词,分别从中国知网(CNKI)和Web of Science(WOS)核心合集数据库中检索近20年的中英文文献。运用Excel、CiteSpace和VOSviewer软件对裸花紫珠中英...目的基于文献计量学方法,分析裸花紫珠的研究现状、热点及发展趋势。方法以裸花紫珠为关键词,分别从中国知网(CNKI)和Web of Science(WOS)核心合集数据库中检索近20年的中英文文献。运用Excel、CiteSpace和VOSviewer软件对裸花紫珠中英文文献的总发文量、期刊发文量、高被引文献、合作网络、关键词等进行可视化分析。结果共纳入文献451篇,其中中文文献397篇,英文文献54篇。裸花紫珠年发文量整体呈上升趋势,发文作者及机构以谷陟欣(海南九芝堂药业有限公司)为主,发文国家以中国为主。刊文量最多的中英文期刊分别为《中草药》(13篇)和Phytochemistry(5篇)。中英文高被引文献主要围绕化学成分、抗炎、抑菌、止血等药理作用及机制研究。裸花紫珠研究热点集中于活性成分、药理作用及机制、临床应用方面。结论裸花紫珠研究在国内取得了一定进展,未来须加强国际合作,其研究趋势可能为深化药理作用及机制、规范临床试验、开发新剂型提供参考。展开更多
本研究以裸花紫珠(Callicarpae Nudiflorae Folium,CNF)的水提取物(CNF water extract,WCNF)和醇提取物(CNF ethanol extract,ECNF)作用于HepG2.2.15细胞,体外评价CNF提取物对HBV的抑制作用,同时结合网络药理学探究CNF抗HBV的潜在作用...本研究以裸花紫珠(Callicarpae Nudiflorae Folium,CNF)的水提取物(CNF water extract,WCNF)和醇提取物(CNF ethanol extract,ECNF)作用于HepG2.2.15细胞,体外评价CNF提取物对HBV的抑制作用,同时结合网络药理学探究CNF抗HBV的潜在作用机制。体外实验采用CCK-8法测定CNF提取物对细胞的毒性;酶联免疫吸附(enzyme-linked immunosorbent assay,ELISA)法检测乙型肝炎病毒e抗原(hepatitis B e antigen,HBeAg)和乙型肝炎病毒表面抗原(hepatitis B surface antigen,HBsAg)分泌量,实时荧光定量PCR(real-time fluorescence quantitative PCR,RT-PCR)法测定HBV DNA及关键靶点基因表达量。通过文献调研、TCMSP数据库检索CNF化学成分、Gene Cards数据库检索HBV相关靶点开展网络药理学研究;运用String数据库构建蛋白互作网络,DAVID数据库进行GO功能及KEGG通路富集分析,Cytoscape软件构建成分-靶点-通路网络可视化并进行拓扑学分析,AutoDock Vina软件进行CNF抗HBV关键成分与核心靶点的分子对接分析,并通过RT-PCR实验对关键靶点进行实验验证。体外实验结果表明CNF水醇提取物均可抑制HbeAg和HBsAg的分泌量,抑制HBV DNA的复制水平,并且水提物的抑制作用明显地强于醇提物。网络药理学结果显示,熊果酸、槲皮素、木犀草素、咖啡酸等化合物为CNF抗HBV的关键活性成分,肿瘤蛋白53(tumor protein 53,TP53)、白介素-6(interleukin-6,IL-6)、肿瘤坏死因子(tumor necrosis factor,TNF)、热休克蛋白90Alpha家族A类成员1(heat shock protein 90 alpha family class A member 1,HSP90AA1)为核心靶点,涉及磷脂酰肌醇-3-激酶/蛋白激酶B(phosphatidylinositol 3-kinase/protein kinase B,PI3K/AKT)、丝裂原蛋白活化激酶(mitogen activated protein kinase,MAPK)等信号通路。此外,实验结果显示CNF提取物可抑制HBV关键基因IL-6、TNF及HSP90AA1表达并提高TP53基因的mRNA表达水平。以上研究揭示了CNF通过多成分、多靶点、多途径发挥抗HBV的作用。后续将以本研究为基础,进一步对其药效物质基础及作用机制进行深入探讨,为CNF抗HBV的临床应用及新药开发提供坚实的理论依据。展开更多
基金supported by the National Natural Science Foundation of China(No.81503224)Guangdong Key Laboratory for translational Cancer research of Chinese Medicine(No.2018B030322011)+1 种基金Department of education of Guangdong Province(No.2016KZD-XM031)Science and Technology Project of Guangzhou City(No.201707010467)
文摘Four new 3,4-seco-labdane diterpenoids,nudiflopenes J-M,were isolated from the leaves of Callicarpa nudiflora along with six known compounds.The structures of these diterpenoids were determined by comprehensive spectroscopic analysis.All the isolated compounds were evaluated for their inhibitory effects on NO production in LPS-stimulated RPMs and RAW264.7 cells.The results suggest that nudiflopenes J-M and other four known compounds showed significant inhibitory effects against NO production comparable to the positive control dexamethasone.
基金supported by the Key Research&Development Plan of Hainan Province(No.ZDYF2018228)Natural Science Foundation of Guangdong Province(No.2016A030313034).
文摘Objective:To evaluate the antidiabetic effect of Callicarpa nudiflora extract in streptozotoc-ininduced diabetic rats.Methods:The chemical constituents in Callicarpa nudiflora extract were identified by UPLCQ-TOF-MS.Callicarpa nudiflora extract(0.15 and 0.3 g/kg)was orally administered to streptozotocin-induced diabetic rats for 42 d.The effects of Callicarpa nudiflora extract on body weight,blood glucose,serum insulin,total cholesterol,triglyceride,LDL-C and HDL-C were investigated,and its effect on liver and pancreatic pathology was assessed by histopathological analysis.Moreover,the expression levels of adenosine 5’-monophosphate-activated protein kinase(AMPK),glucose transporter type 4(GLUT4),phospho-AMPK/AMPK,and p-acetylcoA carboxylase(P-ACC)/ACC in the skeletal muscles and liver were determined by reverse transcription-polymerase chain reaction,Western blotting,and immunohistochemistry.Results:A total of 34 compounds,including 8 iridoids,14 phenylpropanoids,and 12 flavonoids,were identified from Callicarpa nudiflora extract.Callicarpa nudiflora extract significantly reduced blood glucose and significantly restored all other biochemical parameters to near normal levels,including serum insulin,total cholesterol,triglyceride,LDL-C,and HDL-C.Callicarpa nudiflora extract improved insulin resistance and reversed the damage in the liver and pancreas caused by diabetes.Furthermore,Callicarpa nudiflora extract increased the expression levels of phospho-AMPK,GLUT4,P-ACC,and insulin receptor substrate-1 and decreased the expression level of PPAR r in diabetic rats.Conclusions:Callicarpa nudiflora extract improved oral glucose tolerance,lipid metabolism,insulin resistance,and reversed the diabetes-related damage in the liver and pancreas by activating the AMPK-ACC pathway.
基金supported from the China National Key R&D Program(Nos.2017YFE0195100 and 2018YFC.707000)the National Natural Science Foundation of China(Nos.21920102003 and 81903485)。
文摘A phytochemical investigation was carried out on the extract of a medicinal plant Callicarpa nudiflora,resulting in the characterization of five new 3,4-seco-isopimarane(1-5)and one new 3,4-seco-pimarane diterpenoid(6),together with four known compounds.The structures of the new compounds were fully elucidated by extensive analysis of MS,1 D and 2 D NMR spectroscopic data,and time-dependent density functional theory(TDDFT)calculation of electronic circular dichroism(ECD)spectra,and DFT calculations for NMR chemical shifts and optical rotations.
基金Supported by National Key Technology Research and Development Program(SQ2010BAJY1411-07-05)Special Fund for Modernization of Traditional Chinese Medicine in Hainan Province(2015ZY20)+2 种基金Major Science and Technology Project of Hainan Province(ZDZX2013008-01-04)Director Fund of Guangxi Key Laboratory of Functional Phytochemicals Research and Utilization(ZRJJ2014-7,ZRJJ2015-14)Bagui Scholar Program of Guangxi~~
文摘[Objective] This study aimed to study the coagulation effect of Callicarpa nudiflora. [Method] The effects of CaUtcarpa nudiflora on blood coagulation system of mice were investigated by measuring prothmmbin time （PT）, thrombin time（TT）, activated partial thromboplastin time （APTT） and plasma fibrinogen （FIB）, [Result] Compared with the control group, the crude extracts and ：eluted parts of Ca#/carpa nudiflora all significantly shortened PT, APTT and TT, and increased the content of FIB in mice, [Coadadon] The crude extracts and eluted parts of Callicarpa nudiflora have a coagulation-promoting effect through affecting several links of the coagulation process in mice,
基金No: 2018CXTD339, Hainan Natural Science Foundation Innovation Research Team Project, Research on three external preparations of Callicarpa nudiflora.
文摘Objective: To study the effect of the extracts from Callicarpa nudiflora on wound healing and the preparation of ointment. Methods: Totally 50 KM mice were randomly divided into five groups, which were negative control group ,positive control group, high, middle and low dose groups of Callicarpa nudiflora ointment. Mice ear swelling test was conducted to investigate the effect of Callicarpa nudiflora induced by xylene, Bacteriostasis experiments were conducted to observe the bacteriostasis effects of Callicarpa nudiflora on five indicator bacteria, i.e. Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Bacillus subtilis, and Staphylococcus aureus in vitro. A total of 30 new zealand white rabbit with half male and half female were randomly divided into five groups, which were negative control group, positive control group, high, middle and low groups of Callicarpa nudiflora ointment. The rabbits were treated with sterilizing blade and treated with external medicine to observe the effect of the extracts from Callicarpa nudiflora on wound healing. Orthogonal test was used to determine the preparation method of Callicarpa nudiflora ointment,Physical properties, centrifugal stratification test, heat and cold resistance test were conducted. Results:Bacteriostasis experiments in vitro showed high dose group of Callicarpa nudiflora extracts got 16.8 cm antimicrobial zone of Pseudomonas aeruginosa. The experimental results showed that the ear swelling degree of mice induced by xylene was significantly reduced in the middle and high dose groups. All the treatment groups could promote skin healing of rabbit significantly compared with negative control group. There were significantly differences between all the treatment groups and negative control group on the wound healing rate. The optimum preparation process was determined by orthogonal screening test. The optimal formula was as follows: Twain-801.2 g, glycerin monostearate 4.7 g, glycerinum 0.9 g, liquid paraffin 10.8 g, Span801.4 g,solid paraffin 1.4 g,vaseline 0.9 g. The emulsification temperature was 65 ℃,and emulsification time was 25 min. Conclusions: The extract of Callicarpa nudiflora can be used as external preparation and the preparation is in compliance with Chinese pharmacopoeia 2015.
文摘Four new neo-lignans were isolated from the chloroform portion of 70% ethanol extract of the seed crusts of Trewia nudiflora L., and were determined to be 9'-methoxy-7'-en-3',8:4',7-di-epoxyneolignan-3,4,9-triol l, 9'-methoxy-7'-en-3',7:4',8-diepoxyneolignan-3,4,9-triol 2, 9'- ethoxy-7'-en-3',8:4',7-diepoxyneolignan-3,4,9-triol 3, 9'-butoxy-7'-en-3',8:4',7-diepoxy-neolignan -3,4,9-triol 4, respectively, on the basis of spectral data. The antimicrobial bioassays showed that 4 had evident inhibitory activity against Mycobacterium tuberculosis.
基金Innovation research team project of Natural Science Foundation of Hainan Province(No.2018CXTD339)。
文摘Objective:To establish the quality standard of Li folk medicine callicarpa nudiflora powder by modern analytical method.Methods:TLC was used to identify the powder.Five heavy metals and harmful elements(Pb,Cd,As,Hg and Cu)in the powder were determined by flame atomic absorption spectrometry and hydride atomic fluorescence spectrometry.The content of luteolin and Mullein glycoside in the powder was determined by HPLC.Results:the fluorescent spots of TLC identification of powder were clear,and the separation of the two main components was good.It has good specificity,reproducibility and durability,which can be used for the identification of the powder.The inspection items of heavy metals for the identification of powder were established.It was stipulated that:according to the determination method,the lead should not exceed 5 mg/kg;the cadmium should not exceed 0.3 mg/kg;the arsenic should not exceed 2 mg/kg;the mercury should not exceed 0.2 mg/kg;the copper should not exceed 20 mg/kg.The content of luteolin and Mullein glycoside in the powder were determined by HPLC and the content inspection item of the powder was established.It was stipulated that the content of luteolin and Mullein glycoside in the powder should not be less than 0.48%and 5.83%,which can be used for the content limit inspection of the powder.Conclusion:the quality standard method established by our institute is advanced and operable,and can be used for the quality control of the powder.
基金Hainan Natural Science Foundation 2018(No.2018CXTD339)
文摘Objective:To screen the best prescription and technology of Callicarpa nudiflora film,and to study and establish the quality standard of the flim.Methods:The orthogonal test was used to investigate the effect of PVA content,glycerin content,ethanol and water ratio on the comprehensive score of the coating agent;the general inspection method and content determination method were used to control the quality of Callicarpa nudiflora film,and the quality standard of the film was established.Results:The best formulation was selected as polyvinyl alcohol 6.0g,glycerin 2ml,ethanol:water 1:1.The appearance,stability,heat and cold resistance of the film were good,and the quality standard of the film was established.Conclusion:The formulation is simple and feasible,and the quality control method of the film is stable and feasible.
文摘目的基于文献计量学方法,分析裸花紫珠的研究现状、热点及发展趋势。方法以裸花紫珠为关键词,分别从中国知网(CNKI)和Web of Science(WOS)核心合集数据库中检索近20年的中英文文献。运用Excel、CiteSpace和VOSviewer软件对裸花紫珠中英文文献的总发文量、期刊发文量、高被引文献、合作网络、关键词等进行可视化分析。结果共纳入文献451篇,其中中文文献397篇,英文文献54篇。裸花紫珠年发文量整体呈上升趋势,发文作者及机构以谷陟欣(海南九芝堂药业有限公司)为主,发文国家以中国为主。刊文量最多的中英文期刊分别为《中草药》(13篇)和Phytochemistry(5篇)。中英文高被引文献主要围绕化学成分、抗炎、抑菌、止血等药理作用及机制研究。裸花紫珠研究热点集中于活性成分、药理作用及机制、临床应用方面。结论裸花紫珠研究在国内取得了一定进展,未来须加强国际合作,其研究趋势可能为深化药理作用及机制、规范临床试验、开发新剂型提供参考。
文摘本研究以裸花紫珠(Callicarpae Nudiflorae Folium,CNF)的水提取物(CNF water extract,WCNF)和醇提取物(CNF ethanol extract,ECNF)作用于HepG2.2.15细胞,体外评价CNF提取物对HBV的抑制作用,同时结合网络药理学探究CNF抗HBV的潜在作用机制。体外实验采用CCK-8法测定CNF提取物对细胞的毒性;酶联免疫吸附(enzyme-linked immunosorbent assay,ELISA)法检测乙型肝炎病毒e抗原(hepatitis B e antigen,HBeAg)和乙型肝炎病毒表面抗原(hepatitis B surface antigen,HBsAg)分泌量,实时荧光定量PCR(real-time fluorescence quantitative PCR,RT-PCR)法测定HBV DNA及关键靶点基因表达量。通过文献调研、TCMSP数据库检索CNF化学成分、Gene Cards数据库检索HBV相关靶点开展网络药理学研究;运用String数据库构建蛋白互作网络,DAVID数据库进行GO功能及KEGG通路富集分析,Cytoscape软件构建成分-靶点-通路网络可视化并进行拓扑学分析,AutoDock Vina软件进行CNF抗HBV关键成分与核心靶点的分子对接分析,并通过RT-PCR实验对关键靶点进行实验验证。体外实验结果表明CNF水醇提取物均可抑制HbeAg和HBsAg的分泌量,抑制HBV DNA的复制水平,并且水提物的抑制作用明显地强于醇提物。网络药理学结果显示,熊果酸、槲皮素、木犀草素、咖啡酸等化合物为CNF抗HBV的关键活性成分,肿瘤蛋白53(tumor protein 53,TP53)、白介素-6(interleukin-6,IL-6)、肿瘤坏死因子(tumor necrosis factor,TNF)、热休克蛋白90Alpha家族A类成员1(heat shock protein 90 alpha family class A member 1,HSP90AA1)为核心靶点,涉及磷脂酰肌醇-3-激酶/蛋白激酶B(phosphatidylinositol 3-kinase/protein kinase B,PI3K/AKT)、丝裂原蛋白活化激酶(mitogen activated protein kinase,MAPK)等信号通路。此外,实验结果显示CNF提取物可抑制HBV关键基因IL-6、TNF及HSP90AA1表达并提高TP53基因的mRNA表达水平。以上研究揭示了CNF通过多成分、多靶点、多途径发挥抗HBV的作用。后续将以本研究为基础,进一步对其药效物质基础及作用机制进行深入探讨,为CNF抗HBV的临床应用及新药开发提供坚实的理论依据。
