[目的]运用网络药理学及分子对接探讨黄芪散治疗2型糖尿病(T2DM)的作用机制.[方法]通过中药系统药理学分析平台(TCMSP)查找黄芪散组方中黄芪、葛根、桑白皮的的活性成分和作用靶点,通过GeneCards和OMIM数据库查询与T2DM相关的疾病靶点,通过UniProt数据库查询成分和疾病靶点对应的基因名,利用Cytoscape 3.6.1软件构建黄芪散活性成分-靶点网络、蛋白相互作用(PPI)网络,并通过Merge功能构建疾病-成分-靶点网络,筛选出黄芪散治疗T2DM的核心靶点,最后通过DAVID数据库对核心靶点蛋白进行基因本体论(GO)生物过程分析及京都基因与基因组百科全书(KEGG)通路富集分析,预测其作用机制,并绘制GO和KEGG气泡图进行数据可视化.[结果]研究得到包括槲皮素、山柰酚、刺芒柄花素等在内的38个活性成分和ESR1、JUN、PTGS2、AKT1和MAPK1等75个关键靶点.GO功能富集分析得到113个GO条目(P<0.05),主要包括蛋白酪氨酸激酶活性、生长因子活性和核受体活性等生物过程,KEGG富集得到157条信号通路(P<0.05),主要涉及晚期糖基化终末产物(AGEs)-晚期糖基化终末产物受体(RAGE)信号通路、肿瘤坏死因子(TNF)信号通路、磷脂酰肌醇3-激酶(PI3K)/Akt信号通路等.[结论]黄芪散治疗T2DM可能的机制是通过抑制炎症因子分泌,参与抗炎反应,降低氧化应激,激活PI3K/Akt通路等来改善胰岛素抵抗,提高胰岛素敏感性,降低血糖.

目的:研究桑根酮C对游离脂肪酸诱导的人肝癌HepG2细胞脂质蓄积的改善作用.方法:将HepG2细胞分为对照组、模型组、非诺贝特组(10μmol/L)和桑根酮C低、中、高浓度组(2、4、8μmol/L),除对照组外,其余各组细胞加入1 mmol/L游离脂肪酸以诱导建立脂质蓄积模型,且各给药组加入相应含药培养基进行培养.采用油红O染色法观察细胞中脂质蓄积情况,并测定脂质水平和三酰甘油(TG)含量;采用实时荧光定量聚合酶链式反应法和Western blot法检测细胞中过氧化物酶体增殖物激活受体α(PPARα)、肉碱棕榈酰转移酶1(CPT-1)、成脂甾醇元件结合蛋白固醇调节元件结合蛋白1c(SREBP-1c)、脂肪酸合成酶(FAS)、沉默信息调节因子1(SIRT1)、过氧化物酶体增殖物激活受体γ辅助激活物1α(PGC-1α)的mRNA和蛋白表达水平.结果:与对照组比较,模型组细胞核明显萎缩、体积变小,脂滴数明显增加;细胞中脂质水平、TG含量以及SREBP-1c、FAS的mRNA和蛋白表达水平均显著升高(P<0.05或P<0.01),PPARα、CPT-1、SIRT1、PGC-1α的mRNA和蛋白表达水平均显著降低(P<0.01).与模型组比较,桑根酮C各浓度组细胞核萎缩不明显、体积大小正常,脂滴数明显减少;细胞中脂质水平、TG含量以及上述PPARα通路相关基因的mRNA和蛋白(桑根酮C低浓度组的SREBP-1c蛋白除外)表达水平均显著逆转(P<0.05或P<0.01).结论:桑根酮C可改善HepG2细胞的脂质蓄积;其作用机制可能与调节PPARα信号通路、提高细胞脂质氧化能力、抑制脂质合成有关.

The objective of this study was to evaluate the hepatoprotective and metabolic effects of rosmarinic acid (RA) in rats. RA [100 mg/kg body weight (BW)] was intragastrically (i.g.) administered to Sprague-Dawley (SD) rats once a day for seven consecutive days. The rats were then i.g. administered α-naphthylisothiocyanate (ANIT) (80 mg/kg once on the 5th day) to induce acute intrahepatic cholestasis after the last administration of RA. Blood samples were collected at different time points (0.083 h, 0.17 h, 0.33 h, 0.5 h, 0.75 h, 1 h, 1.5 h, 3 h, 4 h, 6 h, 8 h, 12 h, 20 h) after administration, and the levels of RA were estimated by HPLC. Plasma and bile biochemical analysis, bile flow rate, and liver histopathology were measured to evaluate the hepatoprotective effect of RA. The PK-PD curves showed obviously clockwise (AST and ALT) or anticlockwise (TBA, TBIL). Pretreatment with RA at different doses significantly restrained ANIT-induced pathological changes in bile rate, TBA, TBIL, ALT, AST (p < 0.05 or p < 0.01). The relationship between RA concentration and its hepatoprotective effects on acute cholestasis responses was assessed by PK-PD modeling.

目的建立同时测定含中药滴眼液中3种抑菌剂的气相-质谱联用方法。方法采用分时间段SIM监测方式对12种含中药成分的滴眼液中含有的三氯叔丁醇、苯氧乙醇、羟苯乙酯进行检测。DB-5MS毛细管柱(30 mm×0.25mm,0.25μm),进样口温度200℃;炉温80℃保持5 min,10℃/min升温至200℃,保持2 min,40℃/min降温至80℃;柱体积流量1 mL/min;分流比为50∶1。结果 3种抑菌剂在各自的线性范围(0.02~20.0μg/mL,0.05~20.0μg/mL,0.10~10.0μg/mL)内,标准曲线呈良好的线性关系,相关系数均在0.997 2以上。加样回收率位于88.1%~104.1%。结论该方法灵敏度高,为滴眼液的质量监控提供可靠的检测方法。

Previous studies have indicated that borneol has double side effects on the central nervous system (CNS), but the mechanism is unknown. The aim of this study was to clarify the relationship between excitation ratio [contents of excitatory amino acids (AAs) versus that of inhibitory] and the content of natural borneol after a single oral dose. Mice were administered a 1.2 g/kg dose of natural borneol (containing 98% d-borneol) by oral ingestion. Brain samples were collected before administration and at 0.083, 0.167, 0.25, 0.333, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 4 and 5 h after administration. The brain concentration of natural borneol and contents of AA neurotransmitters in mice brain were determined by GC–MS and HPLC-FLU, respectively. After per oral application, natural borneol was absorbed rapidly into the brain and could be determined 5 min after dosing. The maximal brain concentration (86.52 μg/g) was reached after 1 h post-dosing. Natural borneol could affect the contents of AA neurotransmitters in mice brain: l-aspartic acid increased significantly from 0.083 to 1 h after administration, l-glutamic acid increased significantly at 0.333 h and decreased from 1.5 to 5 h, gamma-amino-N-butyric acid increased significantly from 0.167 to 5 h, whereas glycine was not affected. The excitation ratio is the contents of excitatory AAs versus that of inhibitory AAs, which reflects the excitatory or inhibitory state of the body. The excitation ratio elevated transitorily and then declined 0.5 h post-dosing; there were significant differences between 1.5–5 h post-dose compared with pre-dose. The present study indicated that natural borneol could affect the contents of AA neurotransmitters, and the change in excitatory ratio led to borneol’s double side effects on the CNS.

Objective:To explore the influence of cinnabar in Tianwang Buxin Powder(TBP) on the effect of TBP by comparing the effects of TBP with or without cinnabar on the sedation and tranquilizing.Methods:Mice were given TBP and TBP without cinnabar by i.g.,to observe automatic activity,coordinated movement and synergy of sodium pentobarbital with threshold dose and under threshold dose.Results:15 days after administration,automatic activity decreased significantly in high dose group(3 g/kg) of original prescription,sleep duration extended significantly in high dose group of TBP and TBP without cinnabar(2.958 g/kg).There were no significant influence on coordinated movement and synergy of sodium pentobarbital under threshold dose.Conclusion:There was no significant difference on sedation and tranquilizing between TBP and TBP without cinnabar,it is worth to further excavate the medicinal value for toxic Chinese medicine such as cinnabar.

Objective To study the difference of acute toxicity between day and night after oral administration of natural borneol in mice.Methods Mice were randomly divided into day group and night group(n=60),at different time points with different doses of natural borneol,the mortality was observed and the half lethal dose(LD50) was calculated.Results The LD50 of day group was 2 425.9mg/kg,95% confidence interval was 1 939.5~2 940.6 mg/kg;the LD50 of night group was 2 307.3 mg/kg,95% confidence interval was 1 555.6~2 948.2 mg/kg,and the LD50 of day group was larger than night group.Conclusion The acute toxicity is different between day and night for natural borneol in mice,and the toxicity of day group is less than night group.

Objective To study the changes of cerebral borneol content and amino acid neurotransmitters contents in mice after oral administration of natural borneol.Methods Mice were administered natural borneol by intragastric gavage.Before administration and 0.083,0.167,0.25,0.333,0.5,0.75,1,1.5,2,2.5,3 and 4 h after administration,mice brain were sampled to determine borneol content by GC-MS and amino acid neurotransmitters contents by HPLC-FLD.Results Excitation symptoms appeared 5 min after oral administration of natural borneol 1.2 g/kg,lasted 10~15 min and then developed into inhibition.Borneol was detectable in the brain 5 min after administration,reached the the peak at 60 min,the elimination half life was 1.66 h,and the area under the curve was 326.7 μg·h/g.The cerebral content of aspartate(ASP) was increased in mice significantly from 0.083 h to 1 h after administration,the content of glutamic acid(GLU) was increased significantly at 0.333 h and then decreased significantly from 1.5 h to 4 h after administration.The content of γ-aminobutyric acid was(GABA) increased significantly from 0.167 h to 4 h after administration while that of glycine(GLY) was unchanged.Conclusion Natural borneol has two-way actions of excitation and inhibition on central nervous system,and its mechanism is related with the regulation of excitatory / inhibitory neurotransmitters of central nervous system.

Objective To study the changes of the contents of ferulic acid and hydroxysafflor yellow A in the single and mixed decoction of Radix Angelicae Sinensis and Flos Carthami,and to explore the characteristics of Chinese herbs compatibility.Methods A RP-HPLC was performed on a Phenomenex C18 column(4.6 mm×150 mm,5 μm) at 30 ℃.The mobile phase consisted of methanol-0.05 %phosphate solution(25 ∶75 V/V,pH 2.45) at the flow rate of 1.0 mL/min.The automatic sample injector was set at 5 ℃and the ultraviolet detector was operated at 320 nm for ferulic acid and 403 nm for hydroxysafflor yellow A.Results The linearity ranges of ferulic acid was in the range of 0.5 ～10.0 μg/mL(Y=3.8372X+0.0219,r=0.999 8),the average recoveries of adding sample were from 92.68 %to 101.13 %,the RSD(n =6) were less than 3.2 %.The linearity ranges of hydroxysafflor yellow A was in the range of 5.0～100.0 μg/mL(Y=1.3647X-0.3714,r=0.999 9),the average recoveries of adding sample were from 100.17 %to 102.97 %,the RSD(n=6) were less than 2.8 %.The content of ferulic acid in the mixed decoction was 142.64 %higher than that in the single decoction of Radix Angelicae Sinensis,and hydroxysafflor yellow A content was 145.37 %higher than that in the single decoction of Flos Carthami.Conclusion The content of ferulic acid and hydroxysafflor yellow A in mixed decoction is higher than the single decoction of Radix Angelicae Sinensisand and Flos Carthami.The method is simple,fast and accurate.It can be used for the quality control of ferulic acid and hydroxysafflor yellow A simultaneously in extracts of medicinal material.

Aim. To study the pharmacokinetics of dihydroartemisinin (DHA) in Artekin (compound dihydroartemisinin) tablets in Chinese healthy volunteers. Methods. Eighteen healthy volunteers (9 males, 9 females) received Artekin tablets for oral administration. The plasma samples of DHA were analysed by liquid-liquid extraction and determined by HPLC/ESI/MS. Results. The plasma DHA concentration-time curves of single dose and repeated doses of DHA were fitted to a two-compartment open model. The mean pharmacokinetic parameters of DHA in a single dose were: t(1/2(beta))=1.245 +/- 0.495 h, C(max)=243.6 +/- 56.15 microg/l, AUC(0 --> infinity)=450 +/- 69 h x microg/l, V(d)=5.75 +/- 2.2 l/kg and Cl=3.245 +/- 0.38 l/h/kg, while in repeated doses they were: t(1/2(beta))=1.085 +/- 0.298 h, AUC(0 --> infinity)=444.35 +/- 80.43 h x ng/ml, V(d)=4.62 +/- 1.128 ml/kg, Cl=3.0125 +/- 0.875 ml/h/kg, respectively. Conclusion. The study showed that DHA in Artekin was rapidly absorbed, distributed and eliminated in the healthy subjects. The pharmacokinetic properties of DHA in Artekin were not affected by gender in a single dose. While in repeated doses accumulation of DHA did not appear after repeated doses.

The metabolism of borneol is studied by the analysis of incubations of in vitro-prepared rat liver microsomes. A sensitive gas chromatography (GC)-mass spectrometry (MS) method is developed for the identification of borneol and its metabolites. Four novel metabolites, which have not previously been reported, are isolated and confirmed by comparison of the GC-MS method. The biotransformation pathway of borneol in rat liver microsomes is proposed based on the in vitro results.

A sensitive method for the determination of CQP propionic acid in rat plasma was developed and validated after solid-phase extraction. Chromatographic separation was achieved on a reversed-phase Alltima C18 column with the mobile phase of methanol–0.15% (v/v) phosphoric acid solution (pH 2.5) and step gradient elution resulted in a total run time of about 20min. The analytes were detected by using UV detector at 345nm. A good linear relationship was obtained in the concentration range of 50–12,800ng/mL (r=0.9998). The intra-day RSDs and the inter-day RSDs at the concentration of 200, 800, 6400 and 12,800ng/mL were less than 7.0% and 11.0%, respectively. The intra-day accuracy ranged from 96.3 to 106.5% and the inter-day accuracy ranged from 98.6 to 113.4%, respectively. Average extraction recoveries ranged from 83.6 to 94.3% in plasma at the concentrations of 200, 800, 6400 and 12,800ng/mL. This method was successfully applied to the pharmacokinetic studies on rats.

Objective To establish a method for extracting compounds in Fructus Xanthii by accelerated solvent extraction(ASE),and to simultaneously determine the compounds by GC-MS.Methods The extraction was performed by ASE in which the conditions were optimized by orthogonal test,then the compounds in different extracts with different polarity were determined by GC-MS.Results The optimized conditions of ASE for compounds in Xanthium sibiricum fruits were as follows: petroleum ether,ethyl acetate and methanol as extracting solvent in turn in 33 mL extracting tube and the volume of solvent being 65 mL;6 g sample,9 MPa of extracting pressure,90 ℃of extracting temperature,5 min of extracting time,two cycles and one time for each extraction.The GC-MS conditions were as follows: 1 μL extract was analysed by GC-MS(Shimadzu Model QP-2010) instrument fitted with a flame ionisation detector,separation was achieved on a DB-5ms,DB-17ms and DB-WAX column(JW Scientific,30 m,0.25 μm id film thickness,1.4 μm,USA) respectively,the column temperature was maintained at 60 ℃for 3 min and increased to 250 ℃,the scanning range was from m/z 40 to m/z 700(scanning interval: 0.5 s),and the compounds were identified by searching NIST and Wiley registry of mass spectral data.Conclusion There are 61 compounds found in the petroleum ether extract and in which 48 are identified,22 in the ethyl acetate extract and 18 identified,as well as 34 in the methanol extract and 29 identified with the methods of optimized ASE combined with GC-MS.

OBJECTIVE:To study pharmacokinetics of debydroandrographolide, that is a main active component in Chuanxinlian tablet, in healthy Chinese volunteers.METHODS:Eight volunteers were chosen for a single dose of two Chuanxinlian tablet drug concentrations in plasma were measured by HPLC-MS method and the pharmacokinetic parameters were calculated by PK Solution 2.0 software.RESULTS:t(1/2) ka, t(1/2) alpha, t/(1/2) beta, Cmax, tmax and AUCO-t were (0.51 +/- 0.28) h, (0.60 +/- 0.33) h, (3.62 +/- 1.16) h, (147.30 +/- 53.29) microg x L(-1), (1.50 +/- 0.21) h, and (256.63 +/- 64.18) microg x h x L(-1), respectively.CONCLUSION:Dehydroandrographolide has rapid absorption and long elimination rate after oral administration. The results can provide an evidence for the safety and efficiency of Chuanxinlian tablet in clinical application.

The aim of this work was to study the pharmacokinetics of piperaquine in healthy volunteers. Healthy volunteers received piperaquine and tablets of Artekin by oral administration. The plasma samples were analyzed for piperaquine by liquid-liquid extraction and determined by HPLC-UV. The results demonstrated that the plasma drug concentration-time curves of single and multiple dose of piperaquine were fitted to a two-compartment open model. The pharmacokinetics parameters of piperaquine alone in a single dose were: t(1/2(beta))=(317.2-/+126.6)h, AUC(0-->infinity)=(44293-/+12636)h x ng/ml, V(d)=(9490.9-/+2161.9)ml/kg, and Cl=(22.83-/+9.83)ml/h/kg. In Artekin in a single dose these parameters were: t(1/2(beta))=(302.8-/+180.7)h, AUC(0-->infinity)=(46419-/+13670)h x ng/ml, V(d)=(10188.6-/+3520.3)ml/kg, and Cl=(25.48-/+10.89)ml/h/kg, while in Artekin in multiple doses they were: t(1/2(beta))=(298.9-/+101.9)h, AUC(0-->infinity)=(227692-/+56294)h x ng/ml, V(d)=(5031.5-/+1097.8)ml/kg, Cl=(11.91-/+3.046)ml/h/kg, respectively. The absorption and distribution of piperaquine were quick while the elimination was quite slow. There were significant differences in the pharmacokinetics parameters of piperaquine in Artekin between a single dose and multiple doses (p<0.001), suggesting that piperaquine might accumulate in vivo and that attention should be given to its possible adverse drug reactions in clinical treatment.

Objective:To establish a sensitive assay method for determination of piperaquine phosphate in human plasma.Method:Chloroquine phosphate as the internal standard,the plasma was alkalized by 1 mol·L~(-1) sodium hydroxide and extracted with ether,then the organic phase was evaporated to dry thoroughly with soft stream of N_2at 35℃,and the residues were reconstituted with 20μL mobile phase.The reconstitution was chromatographed on the Alltima C_(18) column (4.6 mm×150 mm,5μm)at ambient temperature with mobile phase acetonitrile-0.1% (w/v) trichloroacetic acid solution-phosphoric acid(25:75:0.035,v/v/v),the flow rate was 0.4 mL·min~(-1) and detec- ted at 345 nm.Results:Piperaquine phosphate was well detected in plasma apart from endogenous.The linear range of piperaquine phosphate plasma concentration was 0.02-1.0μg·mL~(-1) (r=0.9993),and the lowest limit of de- tection was 0.01μg·mL~(-1).The method recovery of piperaquine phosphate was 93.4%-106.5% and the extrac- tion recovery was 69.3%-76.3%,the RSDs of intra-day and inter-day were less than 12% for assay respec- tively.Conclusion:The method has good specificity,reproducibility and high sensitivity and it can be applied to de- termine the piperaquine phosphate plasma concentrations accurately.

综述苍耳子的种类、化学成分、药理和毒理作用等研究进展。苍耳子对血糖含量、心血管系统、呼吸系统、免疫系统等具有相应的药理作用;除了富含油脂类化合物之外,苍耳子中还有甾醇、倍半萜内酯和水溶性苷类等,这些化合物可能与上述的药理作用密切相关。由于苍耳子临床应用中会出现不良反应,用药期间应加强临床不良反应监测;同时应对苍耳子具体药理作用的物质基础作进一步研究。

Objective To compare the pharmacokinetics of borneolum syntheticum(BS)and borneol in compound Danshen dropping pills in rats.Method Rats were administrated orally with BS or Danshen dropping pills containing equal amount of borneol in a single dose.The blood contents of borneol at a series of time points were assayed by GC-MS and GC-FID after administration.The drug-time curve was estimated by PK Solution 2.0 pharmacokinetic software(USA).Results The pharmacokinetic parameters are quite different between the single dose of BS and the borneol in the compound Danshen dropping pills:the velocity constants of absorption(2.630h-1),distribution(1.788 h-1)and elimination(0.273 h-1)of BS were much greater than those(1.635h-1,0.784h-1,0.116h-1)of the same quantity of borneol in compound Danshen dropping pills,while the Tmax(0.5 h)of former was shorter than that(0.75 h)in the latter,and the Cmax(16.8 μg/mL)in the former was about half of the latter(31.02 μg/mL).And,at the 18h after administration,the former blood concentration(0.043 μg/mL)was approximately one tenth of the latter.Conclusion It is suggested that we should pay attention to the influence of other ingredients on the main objective component in the study of pharmacokinetics of compound traditional chinese drug.

建立了测定人血浆中的双氢青蒿素的液相色谱-质谱联用法.色谱条件:AlltimaC18柱(2.1×150 mm,5 μm,流动相:甲醇-水(体积比85:15);流速:0.2mL/min;柱温:25℃;进样量:20μL.质谱条件:电喷雾离子源(ESI);用于定量分析的离子分别为m/z307(双氢青蒿素),m/z 275(蒿甲醚).样品用液-液萃取方法处理.双氢青蒿素的线性范围为5～200μg·L-1,定量下限为5μg·L-1,日内、日间精密度(RSD)均小于10%,萃取回收率在71.5%～83.2%之间(n=5),分析方法回收率在98.4%～101.9%之间(n=5).本法操作简便、准确、灵敏度高,适用于双氢青蒿素的药代动力学研究.

Ahighperformance liquid chromatograph(HPLC)withanelectrochemical detector(ECD)and a NUˉCLEOSIL R C 18 column(250mm×4mm,i.d.10μm)was used for the determination of histamine(HA)and5-hydroxytryptamine(5-HT)in rat′s hypothalamus.The mobile phase was a mixture of NaH 2 PO 4 (25mmol/L,containing0.5mmol/LdisodiumEDTAand3mmol/Lsodiuml-octanesulfonate at pH4.8)and acetonitrile in a ratio of100∶14(by volume).The flowrate was0.8mL/min,and the detector voltage was set at0.6V.The homogenate of rat′s hypothalamus was centrifuged at4℃.A part of the supernatant fraction was injected for the direct determination of5-HT,and another part of the supernatant was allowed to react with a derivatizaˉtion reagent before the analysis of HA.Good linearity was found in the range of0.075～1.8μg/mL for HA and0.0625～1.25μg/mL for5-HT.The within-day and intra-day RSDs of the method were≤1.0%and11.4%for HA,and≤3.4%and6.3%for5-HT,respectively.The recoveries of HA and5-HT were94.1%～102.3%and72.8%～89.1%,respectively.The average contents of HA and5-HTin hypothalaˉmus of normal male Sprague-Dawley rats were0.280±0.029μg/mL and0.224±0.068μg/mL,respecˉtively.The method is simple,accurate and rapid,and can be used for the determination of HA and5-HT in rat′s hypothalamus.