Protective mechanism of Paeoniae Radix Alba against chemical liver injury based on network pharmacology,molecular docking,and in vitro experiments

Objective:To explore and validate the potential targets of Paeoniae Radix Alba(P.Radix,Bai Shao)in protecting against chemical liver injury through network pharmacology,molecular docking technology,and in vitro cell experiments.Methods:Network pharmacology was used to identify the common potential targets of P.Radix and chemical liver injury.Molecular docking was used to fit the components,which were subsequently verified in vitro.A cell model of hepatic fibrosis was established by activating hepatic stellate cell(HSC)-LX2 cells with 10 ng/mL transforming growth factor-β1.The cells were exposed to different concentrations of total glucosides of paeony(TGP),the active substance of P.Radix,and then evaluated using the cell counting kit-8 assay,enzyme-linked immunosorbent assay,and western blot.Results:Analysis through network pharmacology revealed 13 key compounds of P.Radix,and the potential targets for preventing chemical liver injury were IL-6,AKT serine/threonine kinase 1,jun protooncogene,heat shock protein 90 alpha family class A member 1(HSP90AA1),peroxisome proliferator activated receptor gamma(PPARG),PTGS2,and CASP3.Gene Ontology(GO)enrichment analysis indicated the involvement of response to drugs,membrane rafts,and peptide binding.Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis revealed that the main pathways involved lipid and atherosclerosis and chemical carcinogenesis-receptor activation.Paeoniflorin and albiflorin exhibited strong affinity for HSP90AA1,PTGS2,PPARG,and CASP3.Different concentrations of TGP can inhibit the expression of COL-I,COL-III,IL-6,TNF-a,IL-1β,HSP-90a,and PTGS2 while increasing the expression of PPAR-γand CASP3 in activated HSC-LX2 cells.Conclusion:P.Radix primarily can regulate targets such as HSP90AA1,PTGS2,PPARG,CASP3.TGP,the main active compound of P.Radix,protects against chemical liver injury by reducing the inflammatory response,activating apoptotic proteins,and promoting the apoptosis of activated HSCs.

Pharmacokinetic and tissue distribution studies of paeoniflorin and albiflorin in rats after oral administration of total glycosides of paeony by HPLC-MS/MS

A simple, sensitive and specific high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS）method combined with solid phase extraction has been developed and validated for the simultaneous quantification of paeoniflorinand albiflorin in rats plasma and tissue homogenate after administration of total glycosides of paeony （TGP）. Chromatographicseparation was achieved on a C15 column （150 mmx4.6 mm, 5 pro） with mobile phase consisted of acetonitrile-0.05% formicacid aqueous （20： 80, v/v） at a flow rate of 0.5 mL/min. The analytes were detected with triple-quadrupole mass spectrometer usingturbo ion spray with negative ionization in the multiple reaction-monitoring （MRM） modes. The results of method validationconformed to the requirements for the determination of biological samples. This method was successfully applied to thepharmacokinetics and tissue distribution study of TGP in rats. The results showed that paeonifiorin and albiflorin wereabsorbed and reached peak value quickly, and had the similar pharmacokinetic characteristics. Both of them underwent a rapid andwide distribution in the tissues throughout the whole body, among which stomach was the main distribution tissue. Moreover, theyhad the ability to cross the blood-brain barrier.

Benefits and Safety of Tripterygium Glycosides and Total Glucosides of Paeony for Rheumatoid Arthritis: An Overview of Systematic Reviews

Objective: To summarize the evidence from systematic reviews(SRs) on the benefits and safety of Tripterygium glycosides(TG) and total glucosides of paeony(TGP), commonly used to treat rheumatoid arthritis(RA) in China, for patients with RA. Methods: SRs of randomized controlled trials(RCTs) on TG or TGP in treating RA were included, by searching 8 databases from their inception until December 2017. Two authors extracted data independently. We assessed the quality of SRs using AMSTAR and graded the quality of evidence according to the GRADE approach. Results: Eleven SRs containing an average of 7.6 RCTs, involving a total of 7,012 participants were included in this overview. On the basis of included SRs, TG and TGP could improve the following indexes for RA patients: American College of Rheumatology(ACR) 20 response rate, ACR50 response rate and ACR70 response rate, swollen joint count, tender joint count, erythrocyte sedimentation rate and C-reactive protein. Moreover, TGP could reduce incidence of hepatotoxicity. The most common adverse effects of TG were gastrointestinal discomfort and gonad toxicity, while for TGP was mild to moderate diarrhea. The overall quality of evidence for these findings ranged from "low" to "moderate". Conclusions: TG and TGP might be 2 potentially effective complementary and alternative drugs for patients with RA. Nevertheless, due to gonad toxicity, TG should only be considered in elderly patients or patients without reproductive needs. More evidence from high quality RCTs and SRs is warranted to support the use of TG and TGP for RA patients.

赤芍总苷药理作用研究进展

赤芍具有清热凉血、散瘀止痛的功效,广泛应用于临床,并于2002年列入可药食同源中药名录。现代研究发现,赤芍总苷系赤芍的主要活性成分,是鸟笼状蒎烷结构的单萜类化合物以及内酯结构的单萜类化合物的总称。10余年来,赤芍总苷不断有新的药理作用发现被报道,但未见系统地总结分析。因此,文章通过归纳国内外相关文献,综述了赤芍总苷自2010年至今被报道的抗肿瘤、抗心脑血管疾病、保肝、抗衰老、改善血流动力学和血液流变学、抗凝、抗血栓以及增强免疫、抗抑郁、抗慢性阻塞性肺疾病、抗溃疡病、抗盆腔炎、舒血管、保护脊髓损伤、保护血管内皮、治疗扁平苔藓及改善糖尿病肾病肾纤维化等药理作用,并提出研究不足与展望,以期为赤芍总苷深入地研究和开发应用提供参考。

Preparation, characterization and pharmacokinetic studies of total paeony glycoside nanocrystals

Total paeony glycoside（TPG） is obtained from Radix Paeoniae Rubra with a variety of bioactivities. However, the low solubility and bioavailability limit its application. The present study aimed to develop TPG nanocrystals to increase the dissolution and then improve the oral bioavailability. TPG nanocrystals were prepared via precipitation and high-pressure homogenization method. The physical-chemical properties of the optimal TPG nanocrystals in terms of particle size, zeta potential, morphology and crystallinity were evaluated. The results showed that TPG nanocrystals had a mean particle size of（210.2±2.5） nm, a polydispersity index of 0.191±0.033 and a zeta potential of（–22.4±1.2） mV. The result of differential scanning calorimetry showed that the nanocrystals were still in crystalline state after the preparation procedure. Transmission electron microscopy（TEM） results showed that the nanosuspension was in spherical shape. The pharmacokinetics of TPG nanocrystals for rats was investigated by liquid chromatography-tandem mass spectroscopy（LC-MS/MS）. Compared with the TPG coarse suspension, TPG nanocrystals exhibited significant increase in AUC0–∞（approximately 1.85-fold）. Taken together, TPG nanocrystals could be used as a promising drug delivery system due to the enhanced oral bioavailability of TPG.

基于Box-Behnken响应面法优化白芍总苷脂质体凝胶的制备工艺

目的:优化白芍总苷脂质体凝胶的制备工艺。方法:以卡波姆用量、甘油用量、三乙醇胺用量为考察因素,均匀度、透明度、流动性为质量评价标准,采用Box-Behnken响应面法优化白芍总苷脂质体凝胶的制备工艺。结果:白芍总苷脂质体凝胶最佳制备工艺的卡波姆用量为2 g,甘油用量为90 g,三乙醇胺用量为30 g。结论:优化后的白芍总苷脂质体凝胶制备工艺制备出的白芍总苷脂质体凝胶外观均匀、透明度高、流动性好。优化后的白芍总苷脂质体凝胶制备工艺稳定可行,可以代替现有的传统制备工艺,应用于临床治疗。

白芍总苷提取和纯化工艺的研究

采用L9（3^4）的正交试验设计，采用水浸提，以加水量、提取时间、提取次数，作为主要考虑因素，确定了白芍总苷的最佳提取工艺为：加入12倍量的水煎煮三次，每次3h。其中提取次数是主要影响因素。在此基础上，采用D101大孔吸附树脂分离纯化，最终白芍中芍药苷转移率可达80．5％。

白芍总甙耐缺氧作用的实验研究

安徽毫县产白芍提取的白芍总甙(TGPs5～40mg/kg)呈剂量依赖性延长小白鼠常压缺氧存活时间。TGPs(20mg/kg)可延长小白鼠减压缺氧存活时间,降低小鼠整体氧耗量,并有降温作用,TGPs(40mg/kg)可降低小白鼠氰化钾中毒性缺氧的死亡率。氯苯吡胺(5mg/kg)可明显拮抗TGPs耐缺氧作用。TGPs(2.5～5mg/kg)小鼠icv可明显延长小鼠常压缺氧存活时间,此作用可被氯苯吡胺拮抗。

赤芍川芎有效部位对兔动脉粥样硬化基质金属蛋白酶的影响

目的观察芎芍胶囊(XSC,赤芍川芎有效部位)对兔动脉粥样硬化模型中基质金属蛋白酶(matrix metalloproteinases,MMPs)表达水平的影响。方法50只新西兰兔随机分为正常对照组、模型对照组、辛伐他汀组、XSC低剂量组和XSC高剂量组,每组10只。正常对照组普通饲料喂养,其余各组高脂饲料喂养,2周后行腹主动脉球囊导管血管损伤术,建立动脉粥样硬化模型,继续高脂喂养,辛伐他汀组给予辛伐他汀2.5mg/(kg·d),XSC低、高剂量组分别给予芎芍胶囊0.24g/(kg·d)和0.48g/(kg·d),连续给药6周后,各组采血测血脂;酶联免疫法测定血清MMP-3、MMP-9和MMP组织抑制因子-1(TIMP-1)的表达;免疫组化方法检测斑块组织MMP-3及白细胞分化抗原40配体(CD40L)的蛋白表达。结果与模型对照组比较,辛伐他汀组、XSC高、低剂量组血清MMP-3、MMP-9水平显著下降(P<0.05),同时斑块内MMP-3和CD40L表达也明显降低(P<0.05,P<0.01);辛伐他汀组和XSC高、低剂量组较模型对照组对血清总胆固醇均有不同程度的下降(P<0.05,P<0.01),辛伐他汀组还显著降低血清甘油三酯和低密度脂蛋白胆固醇;各组TIMP-1变化差异无统计学意义(P>0.05)。结论赤芍川芎有效部位具有一定稳定斑块的作用,抑制动脉粥样硬化模型兔血管壁中和血清MMP-3、MMP-9及相关因子表达,可能是其可能机制之一。

赤芍总苷对小鼠学习记忆能力的改善作用

目的 观察赤芍总苷对小鼠学习记忆能力的影响。方法 选用东莨菪碱所致小鼠记忆获得 ,环己酰亚胺所致记忆巩固 ,乙醇所致记忆再现障碍模型及戊巴比妥钠所致空间分辨障碍模型。结果 赤芍总苷显著减少上述记忆障碍小鼠受刺激时间 ,延长平台停留期 ,减少错误次数 ,并调节反应潜伏期 ;对空间分辨障碍小鼠 ,赤芍总苷具有一定增加达岸成功率 ,减少错误次数及缩短达岸时间的作用。

赤芍总甙活血化瘀作用的研究

观察了赤芍总甙的活血化瘀作用。研究结果表明,赤芍总甙可以显著改善机体微循环状态,降低血清、血浆粘度,抑制ADP诱导的血小板聚集,延长凝血酶原时间(PT)和活化部分凝血活酶时间(KPTT)。

赤芍总甙对原代培养大鼠神经细胞损伤模型的保护作用

目的探讨赤芍总甙对大鼠神经细胞脑缺血样损伤模型的保护作用及其机制。方法采用组织培养法 ,以原代大鼠大脑皮层神经细胞为材料 ,制备缺糖损伤、缺氧损伤、自由基损伤、咖啡因损伤、一氧化氮损伤及NMDA毒性损伤模型。结果形态学检查发现赤芍总甙 (totalpaeonyglycoside,TPG)对6六种脑缺血样损伤模型中的大鼠神经细胞具有明显保护作用 ,结晶紫染色也提示TPG可显著提高损伤模型中神经细胞的存活数。

赤芍总苷对D-半乳糖衰老小鼠学习记忆及代谢产物的影响

目的 观察赤芍总苷对D 半乳糖所致衰老小鼠学习记忆及与神经介质调节相关酶及代谢产物的影响。方法 选用长期皮下注射D 半乳糖形成的衰老小鼠模型。结果 赤芍总苷对衰老小鼠的学习记忆障碍有显著的改善作用 ,穿梭试验中小鼠主动回避潜伏期和主动回避次数明显增加 ;水迷宫试验发现 ,赤芍总苷可有效提高小鼠的达岸正确率 ,减少小鼠的错误次数。赤芍总苷可显著降低衰老小鼠脑组织中单胺氧化酶 (MAO)水平 ,提高胆碱酯酶 (CHE)活性 ;并抑制脂质过氧化产物丙二醛 (MDA)含量的增加 ,提高脑组织超氧化物岐化酶 (SOD)水平。此外 ,赤芍总苷对衰老小鼠免疫功能低下及脑萎缩状态有显著的改善。

赤芍总苷对培养乳鼠心肌细胞损伤的抗氧化作用

目的 探讨赤芍总苷对心肌细胞损伤的抗氧化保护作用。方法 以培养的乳鼠心肌细胞加入异丙基肾上腺素造成缺血缺氧损伤模型 ,对比分析正常对照组、药物损伤组、辅酶Q10 阳性对照组、以及不同剂量赤芍总苷保护组培养液中超氧化物歧化酶、丙二醛和一氧化氮含量。结果 损伤组总SOD、CuZn SOD和Mn SOD水平均明显下降 ,MDA和NO含量显著升高 ;而辅酶Q10 组和赤芍总苷组上述指标都有不同程度的改善 ,其中大剂量赤芍总苷组的保护作用接近或优于阳性对照组。结论 赤芍总苷对异丙基肾上腺素造成的心肌损伤具有保护作用 ,且呈现剂量依赖关系 ,作用机制与增强细胞抗氧化作用 ,减少自由基和脂质过氧化物导致的细胞膜损伤有关。

白芍总苷对雷公藤多苷治疗狼疮性肾炎减毒增效作用的实验研究

目的探讨白芍总苷(TGP)对雷公藤多苷(TG)治疗狼疮性肾炎(lupus nephritis,LN)小鼠的减毒增效作用。方法取B6D2F1代杂交小鼠建立慢性移植物抗宿主病(cGVHD)LN小鼠模型,将成模小鼠随机分为3组,模型(LN)组、TG组、TG+TGP组,每组6只,另设正常对照组6只。TG组给予雷公藤多苷灌胃,剂量为30mg·kg-1·d-1;TG+TGP组给予雷公藤多苷及白芍总苷灌胃,剂量分别为30mg·kg-1·d-1和200mg·kg-1·d-1,正常对照组和LN组分别给予等容量生理盐水灌胃,每日给药1次,各组给药方法及时间均相同,共计4周。实验结束时,观察并比较各组小鼠的24h尿蛋白排泄率(UPE)、肝肾功能及肾脏病理改变。结果与LN组相比,TG组、TG+TGP组小鼠的24hUPE、尿素(BUN)、肌酐(SCr)显著降低(P<0.05或P<0.01),TG+TGP组显著低于TG组(P<0.05或P<0.01)。与LN组相比,TG组小鼠的ALT、AST和MDA均显著增高(P<0.01),SOD、GSH均显著降低(P<0.05或P<0.01)。与TG组相比,TG+TGP组的ALT、AST、MDA均显著降低(P<0.01),SOD、GSH均显著增高(P<0.01)。TG+TGP组对LN肾脏病理改变的改善作用显著优于TG组(P<0.05或P<0.01)。结论TGP对TG治疗LN具有减毒增效的作用。

西洋参赤芍配伍对大鼠心肌梗死后早期心室重构心肌纤维化的影响

目的观察益气活血配伍(西洋参茎叶总皂苷+赤芍总苷)对大鼠心肌梗死后早期心室重构炎性因子和心肌纤维化的影响,并从胶原代谢方面探讨可能的作用机制。方法结扎Wistar大鼠冠状动脉前降支制备AMI模型,将90只造模成功的大鼠随机分为6组,每组15只,分别为假手术组(冠状动脉前降支仅穿线不结扎)、模型组、阳性对照组(灌胃缬沙坦7.2 mg·kg^(-1)·d^(-1))、益气组(灌胃西洋参茎叶总皂苷162 mg·kg^(-1)·d^(-1))、活血组(灌胃赤芍总苷54 mg·kg^(-1)·d^(-1))和益气活血组(灌胃西洋参茎叶总皂苷162 mg·kg^(-1)·d^(-1)+赤芍总苷54mg·kg^(-1)·d^(-1))。第2天开始灌胃给药,连续4周,假手术组和模型组灌胃等量蒸馏水。4周后取材,超声心动图检测心脏结构及心功能变化,HE染色观察心肌组织病理变化,免疫组织化学染色法检测缺血心肌Ⅰ、Ⅲ型胶原蛋白的表达。放射性免疫法检测血清心肌损伤标记物B型脑钠尿肽(BNP)、肌钙蛋白T(cTnT),炎症因子肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β),纤维化指标层连蛋白(LA)、透明质酸(HA)、Ⅲ型前胶原氨基末端肽(PⅢNP)的含量。结果与模型组比较:(1)益气活血组大鼠心肌细胞炎性浸润减少,心肌纤维间隙水肿和心肌纤维细胞坏死变性明显减轻;(2)阳性对照组、活血组和益气活血组大鼠左室舒张末期内径(LVDd)均显著降低,左室射血分数(LVEF)均显著升高(P<0.05,P<0.05),益气活血组左室收缩末期内径(LVDs)显著降低(P<0.05);(3)阳性对照组和益气活血组心肌损伤标记物血清BNP水平明显降低(P<0.05);(4)阳性对照组和益气活血组炎性因子血清TNF-α、IL-1β含量显著降低(P<0.05,P<0.01);(5)益气活血组纤维化指标血清LA、HA、PⅢNP含量明显降低(P<0.05);(6)阳性对照组和益气活血组损伤心肌Ⅰ、Ⅲ型胶原蛋白表达明显降低(P<0.05,P<0.01)。结论益气活血配伍可以减轻心梗后心肌纤维化过程中的炎性反应,减少损伤心肌纤维化因子的释放及胶原的堆积从而达到减轻心室重构改善心功能的作用。

赤芍总甙对实验性脑缺血的保护作用

目的:观察赤芍总甙对小鼠实验性脑缺血的影响。方法:选用小鼠断头模型及结扎双侧颈总动脉形成的小鼠脑缺血模型。结果:赤芍总甙明显延长小鼠断头后张口喘气时间,减少脑含水量,并可有效降低脑毛细血管通透性。结论:赤芍总甙对小鼠急性实验性脑缺血具有显著的保护作用。

白芍与柴胡不同比例配伍白芍总苷及苯甲酸水煎出量的比较

[目的]考察白芍与柴胡不同比例配伍共煎芍药总苷及苯甲酸的煎出量,以探讨传统药对白芍配伍柴胡对有效成分及有害成分煎出的影响。[方法]单味白芍、白芍与柴胡配伍(1∶2,1∶1,2∶1),水煎提取,分别直接分离和水解后再分离测定游离苯甲酸及芍药总苷,采用高效液相色谱法(HPLC)测定。[结果]白芍配伍柴胡,游离苯甲酸煎出量均比白芍单煎明显降低,芍药总苷煎出量与白芍单煎比变化不大。[结论]白芍配伍柴胡合煎与白芍单煎相比,可显著降低有害成分苯甲酸的煎出量。

白芍总甙对大鼠佐剂性关节炎及其免疫功能的影响

本文研究了白芍总甙(TGP)对大鼠佐剂性关节炎(AA)的治疗作用及其机理。TGP 50 mg·kg^1·d^1 ig,连续11d对大鼠多发性关节炎有明显的防治作用,同时可使AA大鼠腹腔巨噬细胞升高的H_2O_2和白细胞介素(IL-1)水平下降,并可使AA大鼠低下的胸腺细胞有丝分裂原反应及脾淋巴细胞产生IL-2能力恢复正常。表明TGP对AA大鼠有抗炎和机能依赖性的免疫调节作用。

白芍总苷和维生素B2治疗复发性阿弗他溃疡的疗效评定

目的:观察与比较白芍总苷和维生素B2在治疗复发性阿弗他溃疡(recurrent aphthous ulcer,RAU)中的临床疗效及不良反应。方法:将100例RAU患者随机分为2组,治疗组50例采用白芍总苷治疗,对照组50例采用维生素B2治疗,2组疗程均为6个月。根据中华口腔医学会黏膜病专业委员会的"RAU疗效评价试行标准"进行疗效评价。结果:2组患者在治疗期内的总间歇期均延长,溃疡总数目均减少。2组比较,白芍总苷组疗效优于维生素B2组(P<0.05);白芍总苷对RAU间歇期的延长优于维生素B2(t=4.89,P<0.01);对RAU溃疡数目的减少也优于维生素B2(t'=7.87,P<0.01)。白芍总苷组9例发生轻度不良反应,维生素B2组未观察到不良反应。结论:白芍总苷治疗RAU具有较好的疗效不良反应轻微。