Determination of Benzoylaconitine Compounds in the Decoctions of Aconiti Lateralis Radix Praeparata(Heishun Pieces),Trichosanthis Fructus and Their Combination

[Objectives]This study was conducted to determine the contents of benzoylmesaconine,benzoylaconitine and benzoylhypacoitine in the decoctions of Heishun pieces,Trichosanthis Fructus and their combination.[Methods]Heishun pieces,Trichosanthis Fructus and their combination were extracted for different time periods,and then grouped.HPLC was performed using an Agilent ZORBAX SB-C 18 chromatographic column(4.6 mm×250 mm,5μm)and acetonitrile-0.02 mol/L sodium dihydrogen phosphate as the mobile phase at a flow rate of 1 mL/min and a column temperature of 30℃,and the sample volume was 20μL.The detection wavelength was 230 nm.[Results]The total amounts of benzoylmesaconine,benzoylaconitine and benzoylhypacoitine in the single decoction group of Heishun pieces were all significantly different from those in the combined decoction group at corresponding time.[Conclusions]The total content of the benzoylaconitine type increased significantly after the combined decoction of Heishun pieces and Fructus Trichosanthis,which proves the scientificity of"eighteen incompatible medicaments,19 counteraction"in traditional Chinese medicine to some extent.

Protective Effects of Zingiberis and Acniti Praeparatae Decoction on Myocardial IschemiaReperfusion Injury in Rats

This study aimed to investigate the protective effects of zin-giberis and acniti praeparatae decoction on oxidative stress injury induced by my-ocardial ischemia reperfusion in rats. [Method] Myocardial ischemia-reperfusion was performed by ligation of the left anterior descending coronary artery for 30 min, fol-lowed by reperfusion for 60 min. The effects of zingiberis and acniti praeparatae decoction on ECG ST segment, myocardial infarction percentage, malondialdehyde （MDA） content in the serum, superoxide dismutase （SOD） activity and other indica-tors were observed. [Result] Zingiberis and acniti praeparatae decoction could effec-tively inhibit ECG ST segment elevation caused by myocardial ischemia-reperfusion injuries, reduce the percentage of myocardial infarction, decline the content of MDA in the serum, and increase the activity of SOD. [Conclusion] Zingiberis and acniti praeparatae decoction exhibits protective effects on oxidative injuries caused by my-ocardial ischemia-reperfusion injuries in rats, which may be involved in reducing the formation of myocardial free radicals and enhancing antioxidant capacity of my-ocardium.

Radix Polygoni Multiflori Praeparata and Dioscorea Bulbifera Rhizome Decoctions Display Combined Effects Detected by a Three-Probe Drug Cocktail with Substrates of Rat Hepatic Cytochrome P450 Enzymes

Objectives: Radix Polygoni Multiflori Praeparata (RPMP) and Dioscorea Bulbifera Rhizomes (DBR) are used in Chinese herbal medicine and have been frequently reported for adverse reactions on liver. In this research, we aimed to evaluate in vivo effects of RPMP and DBR on rat cytochrome P450 enzymes (CYP1A2, CYP2E1 and CYP3A2) with their respective substrates as probes. Methods: Rats were orally administered RPMP, DBR and RPMP/DBR combination at 12, 10 and (12 + 10) g/kg, respectively, or saline as a control, once daily for 7 days. Thereafter, a cocktail containing 10 mg/kg caffeine, 20 mg/kg chlorzoxazone and 10 mg/kg dapsone was tail vein injected to rats. At defined time points, plasma drug concentrations were simultaneously evaluated by HPLC. Pharmacokinetic parameters simulated by DAS software were used to assess RPMP and/or DBR effects on cytochrome P450 enzymes activity. ANOVA and Dunnett’s test were used for data analysis. Results: Caffeine metabolism was enhanced in RPMP animals and reduced after pretreatment with DBR, but no effect was observed in RPMP/DBR combination group. Chlorzoxazone and dapsone metabolism was enhanced in both RPMP and DBR groups and consequently in combination group. The data suggested that RPMP independently induces rat CYP1A2, CYP2E1 and CYP3A2 activity, while DBR independently inhibits activity of rat CYP1A2 and induces that of CYP2E1 and CYP3A2. RPMP/DBR combination showed no significant benefit compared with the two drugs alone and even showed a neutralized effect in CYP1A2 activity. Conclusions: Caution is needed when RPMP and/or DBR are co-administered with drugs metabolized by human CYP1A2, CYP2E1 and CYP3A4.

Research Progress in Toxicity and Toxicity-reducing and Efficacy-enhancing Compatibility of Radix Aconiti Lateralis Praeparata

Radix Aconiti Lateralis Praeparata contains alkaloids,flavonoids,polysaccharides,organic acids and organic alkalis,among which diester alkaloids are important effective ingredients,which can be used for treating heart failure,rheumatic heart disease,coronary heart disease,hypotension,shock and other diseases.However,it also contains the main ingredients that produce toxic effects and are highly toxic to the cardiovascular system,nervous system,digestive system,reproductive system,embryos,and livers and kidneys.In this study,we analyzed the toxicity-reducing and efficacy-enhancing effects of the compatibility of Radix Aconiti Lateralis Praeparata with with single herbs Glycyrrhiza uralensis Fisch.,Panax ginseng C.A.Meyer,Zingiber ojjicinale Rosc.and Rheum palmatum L.,aiming to provide references for further exploring the appropriate compatibility conditions of traditional Chinese medicine in the future and improving the safety of the clinical application of Radix Aconiti Lateralis Praeparata.

Deciphering the potential mechanism of Radix Rehmanniae Praeparata for treating cancer-related anemia based on network pharmacology and molecular docking technology

Background:Rehmanniae Radix Praeparata(RRP,Shu Dihuang in Cinese)is a traditional Chinese herb with multiple pharmacological effects and is commonly used to treat blood deficiency syndrome,such as cancer-related anemia(CRA),alone or in combination with other herbs.However,its main active ingredients and mechanisms of action in treating CRA remain unknown.This study aims to elucidate RRP’s potential mechanism and main active components in treating CRA by using network pharmacology and molecular docking technology system.Methods:The main components of RRP were obtained by the TCMSP database and literature search,and active components and potential targets were obtained by the SwissADME and SwissTargetPridiction databases.CRA targets were collected through GeneCards,DisGeNET,and DrugBank databases.Protein-protein interaction networks of potential targets were constructed via STRING 11.5 and analyzed visually with Cytoscape 3.9.1.The Metascape platform was used for Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis,which were subsequently visualized with Cytoscape 3.9.1 or SangerBox platform.Moreover,Autodock Vina was used for the molecular docking of potential targets and main active ingredients that were visualized with PyMOL software.Results:In this study,31 main active ingredients of PPR were screened,and 155 related targets related to CRA were unearthed.Protein-protein interaction results showed that PPR’s core proteins for CRA intervention correlate to STAT3,SRC,MAPK3,MAPK1,PIK3R1,PIK3CA,and AKT.Multiple signaling pathways were closely related to the treatment of CRA intervened by PPR,including the PI3K-Akt signaling pathway,HIF-1 signaling pathway,JAK-STAT3 signaling pathway,TNF-αsignaling,cytokine signaling pathway and NF-kappB signaling pathway,which are closely involved in the proliferation and differentiation of hematopoietic stem cell and inflammatory response.Molecular docking results showed that these potential targets had good conformation with the core active components of RRP for treating CRA.Conclusion:This study revealed RRP’s main active components and potential molecular mechanisms in treating CRA,providing a reference for subsequent basic research.

Rehmanniae Radix Praeparata aqueous extract improves hepatic ischemia/reperfusion injury by restoring intracellular iron homeostasis

Hepatic ischemia/reperfusion injury(HIRI)is a common pathophysiological condition occurring during or after liver resection and transplantation,leading to hepatic viability impairment and functional deterioration.Recently,ferroptosis,a newly recognized form of programmed cell death,has been implicated in IRI.Rehmanniae Radix Praeparata(RRP),extensively used in Chinese herbal medicine for its hepatoprotective,anti-inflammatory,and antioxidant properties,presents a potential therapeutic approach.However,the mechanisms by which RRP mitigates HIRI,particularly through the regulation of ferroptosis,remain unclear.In this study,we developed a HIRI mouse model and monocrotaline(MCT)-and erastin-induced in vitro hepatocyte injury models.We conducted whole-genome transcriptome analysis to elucidate the protective effects and mechanisms of RRP on HIRI.The RRP aqueous extract was characterized by the presence of acteoside,rehmannioside D,and 5-hydroxymethylfurfural.Our results demonstrate that the RRP aqueous extract ameliorated oxidative stress,reduced intracellular iron accumulation,and attenuated HIRI-induced liver damage.Additionally,RRP significantly inhibited hepatocyte death by restoring intracellular iron homeostasis both in vivo and in vitro.Mechanistically,the RRP aqueous extract reduced intrahepatocellular iron accumulation by inhibiting ZIP14-mediated iron uptake,promoting hepcidin-and ferroportin-mediated iron efflux,and ameliorating mitochondrial iron aggregation through upregulation of Cisd1 expression.Moreover,siRNA-mediated inhibition of hamp synergistically enhanced the RRP aqueous extract's inhibitory effect on ferroptosis.In conclusion,our study elucidates the mechanisms by which RRP aqueous extracts alleviate HIRI,highlighting the restoration of iron metabolic balance.These findings position RRP as a promising candidate for clinical intervention in HIRI treatment.

A feasible,economical,and accurate analytical method for simultaneous determination of six alkaloid markers in Aconiti Lateralis Radix Praeparata from different manufacturing sources and processing ways

Aconiti Lateralis Radix Praeparata(Fuzi) is a commonly used traditional Chinese medicine in clinic for its potency in restoring yang and rescuing from collapse. Aconiti alkaloids, mainly including monoester-diterpenoidaconitines(MDAs) and diester-diterpenoidaconitines(DDAs), are considered to act as both bioactive and toxic constituents. In the present study, a feasible, economical, and accurate HPLC method for simultaneous determination of six alkaloid markers using the Single Standard for Determination of Multi-Components(SSDMC) method was developed and fully validated. Benzoylmesaconine was used as the unique reference standard. This method was proven as accurate(recovery varying between 97.5%-101.8%, RSD < 3%), precise(RSD 0.63%-2.05%), and linear(R > 0.999 9) over the concentration ranges, and subsequently applied to quantitative evaluation of 62 batches of samples, among which 45 batches were from good manufacturing practice(GMP) facilities and 17 batches from the drug market. The contents were then analyzed by principal component analysis(PCA) and homogeneity test. The present study provided valuable information for improving the quality standard of Aconiti Lateralis Radix Praeparata. The developed method also has the potential in analysis of other Aconitum species, such as Aconitum carmichaelii(prepared parent root) and Aconitum kusnezoffii(prepared root).

Correlation between 5-hydroxymethylfurfural content and color of Rehmanniae Radix and Rehmanniae Radix Praeparata

In the present study,we explored the correlation between 5-hydroxymethylfurfural(5-HMF)and color of Rehmanniae Radix(RR)and Rehmanniae Radix Praeparata(RRP).The color was observed by human eyes under sunlight.The chromatic value and color difference were detected by a colorimeter.The water content was determined by an oven-drying method.The 5-HMF content was determined by an HPLC method.The correlation between the 5-HMF content and the color was analyzed by bivariate correlation analysis.The results showed that the color was greyish-brown and dark brown for RR,and it was black for RRP.The deepening color of RRP was reflected in the decrease of L^*,a^*and b^*values.The water content was in accord with the requirement of Chinese Pharmacopoeia.The 5-HMF contents of RR and RRP were 0.9711–25.71μg/g and 213.4–4010μg/g,respectively.The average 5-HMF contents of RR and RRP were 8.059μg/g and 1433μg/g,respectively.The 5-HMF content in RRP was higher than that in RR.The correlations between 5-HMF content and L^*,a^*and b^*values were significant.The color of RRP was deeper than that of RR.The 5-HMF content of RRP was higher than 0.02%,while it was less than 0.02%of RR,which could be used as a marker component for judging RR and RRP.

Hematopoietic effect of small molecular fraction of Polygoni multiflori Radix Praeparata in cyclophosphamide-induced anemia mice

The aim of this study is to investigate the protective effects of a small molecular fraction(SMF) of Polygoni multiflori Radix Praeparata(PMRP) in a cyclophosphamide(CTX) induced anemia mouse model. Small molecular fraction of PMRP was prepared and identified by high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF-MS). In pharmacology, we examined the peripheral hemogram and thymus and spleen index. The content of granulocyte-macrophage colony-stimulating factor(GM-CSF) in serum was mensurated by enzyme-linked immunosorbent assay(ELISA);The level of superoxide dismutase(SOD), catalase(CAT), total antioxidant capacity(T-AOC), and malondialdehyde(MDA) in serum and spleen tissue homogenate were detected, and glutathione peroxidase(GSH-PX) was assayed in spleen. The results show that SMF can significantly accelerate the recovery of peripheral hemogram, increase the activity of antioxidant enzymes and GM-CSF in serum and spleen. SMF also increases the number of spleen cells, improves bone marrow pathology. In conclusion, the SMF of PMRP promoted the recovery of hematopoietic function in a CTX-induced anemia mouse, which can support SMF to be used as an adjunct to chemotherapy to counteract its side effects.

Inhibitory effect of Sumu(Lignum Sappan) plus Fuzi(Radix Aconiti Lateralis Praeparata) on a lung carcinoma model

OBJECTIVE: To investigate the inhibitory effect of Sumu(Lignum Sappan) plus Fuzi(Radix Aconiti Lateralis Praeparata)(SF) on the growth and metastasis of Lewis lung carcinoma.METHODS: A lung carcinoma model was established by subcutaneously inoculating Lewis lung carcinoma cells into C57 BL/6 mice. The mice were randomly divided into four groups of 13 mice(control, low-dose, moderate-dose, and high-dose), and gavaged once-daily for 21 consecutive days. The rates of tumor inhibition, metastasis, and metastasis inhibition were observed. The differential expressions of s P-selectin and vascular endothelial growth factor C(VEGFC) were compared between the treatment groups and the control group.RESULTS: The tumor weights differed significantly between the treatment groups and the control group(P < 0.05). Administration of SF in the moderate-dosage and low-dose groups significantly inhibited the expression of s P-selectin and VEGFC(both P < 0.05), suggesting anti-tumor activity.CONCLUSION: SF can inhibit the growth and metastasis of Lewis lung carcinoma.

Study on Hot Property of Aconiti Lateralis Radix Praeparata and Its Compatibility with Zingiberis Rhizoma Based on Animal Temperature Tropism

Objective To establish an objective method for evaluating the intrinsic characteristics between hot and cold properties of herbal drugs and to study the hot property of Aconiti Lateralis Radix Praeparata(ALRP) and its compatibility with Zingiberis Rhizoma(ZR) on animal temperature tropism.Methods The equipment with cold/hot pads was used to investigate the variety of temperature tropism among mice treated by ALRP and its compatibility with ZR.Meanwhile,the activities of adenosine triphosphatase(ATPase),total anti-oxidant capability(T-AOC),and total superoxide dismutase(T-SOD) were measured.Results Compared with the homologous ALRP group,the compatibility between ALRP and ZR(AZ) had a stronger action against the decreasing of remaining rate on hot pad(P < 0.05),suggesting an enhancement of cold tropism.Meanwhile,the internal indexes of Na+-K+-ATPase activity,Ca2+-Mg2+-ATPase activity,and oxygen consuming volume increased significantly(P < 0.05),suggesting an enhancement of energy metabolism.The changes of T-AOC and T-SOD activity suggested that AZ reinforced the anti-oxidative capability of mice.In addition,it could be inferred from cluster analysis that the activity of AZ was concretely stronger than that of homologous ALRP.Conclusion ALRP and ZR are both herbs with hot property,but AZ has an enhanced trend of hot property.This external behavior of cold tropism and the internal activity of energy metabolism and anti-oxidant might reflect the internal hot property in an intuitive and objective way.The changes of ATPase activity of liver tissue might be the mechanism of drug action.

Hot Property of Aconiti Lateralis Radix Praeparata and its Compatibility with Zingiberis Rhizoma Based on Animal Temperature Tropism

Abstract: Objective To establish an objective method for evaluating the intrinsic characteristics between hot and cold properties of herbal drugs and to study the hot property of Aconiti Lateralis Radix Praeparata (ALRP) and its compatibility with Zingiberis Rhizoma (ZR) on animal temperature tropism. Methods The equipment with cold/hot pads was used to investigate the variety of temperature tropism among mice treated by ALRP and its compatibility with ZR. Meanwhile, the activities of adenosine triphosphatase (ATPase), total anti-oxidant capability (T-AOC), and total superoxide dismutase (T-SOD) were measured. Results Compared with the homologous ALRP group, the compatibility between ALRP and ZR (AZ) had a stronger action against the decreasing of remaining rate on hot pad (P 【 0.05), suggesting an enhancement of cold tropism. Meanwhile, the internal indexes of Na+-K+-ATPase activity, Ca2+-Mg2+-ATPase activity, and oxygen consuming volume increased significantly (P 【 0.05), suggesting an enhancement of energy metabolism. The changes of T-AOC and T-SOD activity suggested that AZ reinforced the anti-oxidative capability of mice. In addition, it could be inferred from cluster analysis that the activity of AZ was concretely stronger than that of homologous ALRP. Conclusion ALRP and ZR are both herbs with hot property, but AZ has an enhanced trend of hot property. This external behavior of cold tropism and the internal activity of energy metabolism and anti-oxidant might reflect the internal hot property in an intuitive and objective way. The changes of ATPase activity of liver tissue might be the mechanism of drug action.

Screening and Identifying Hepatotoxic Components in Polygoni multiflori Radix and Polygoni multiflori Radix Praeparata

Objective: In this study, the hepatotoxic components of Polygoni multiflori Radix and Polygoni multiflori Radix Praeparata(known as Heshouwu [HSW] and Zhiheshouwu [ZHSW] in China, respectively) were screened, isolated, and identified. Materials and Methods: The ethanol extracts of HSW and ZHSW were separated into 80 fractions according to their polarity in the preparation liquid phase. Chang liver cell line was used to screen the toxic components of HSW and ZHSW in vitro. The obtained toxic mixture was further collected, isolated, and identified to confirm the hepatotoxic compounds of HSW and ZHSW. Results: The identifid hepatotoxic compounds include 2,3,5,4’-tetrah ydroxystilbene-2-O-β-D-glucoside, emodin, physcion-8-O-β-d-glucoside, physcion, and citreorosein, the fist two among them were the main components of HSW and ZHSW. After processing of HSW, the contents of 2,3,5,4’-tetrahydroxystilbene-2-O-β-D-glucoside and emodin in ZHSW were signifiantly decreased. Conclusions: The traditional processing with herb has signifiant effects on the components, especially the toxic components, in the extract of HSW and is an effective method to reduce its toxicity.

陕产附子炮制前后急性毒性及强心作用研究

目的:考察陕产附子炮制前后的急性毒性和强心作用。方法:通过对生附子LD_(50)的测定和自制黑顺片最大耐受量实验比较生附子和自制黑顺片的急性毒性;通过复制慢性心衰大鼠模型,检测大鼠体质量、心脏指数、血流动力学指标,检测血清BNP及cTnI含量以及组织病理学检查比较其强心作用。结果:生附子的半数致死量(LD_(50))为9.45 g/kg,黑顺片的最大耐受量为27.39 g/kg,经炮制后毒性降低。与模型组相比,自制黑顺片组大鼠症状及体征均有改善,能够降低心脏指数、左心室指数降低和左心室舒张压(LVEDP),升高左心室收缩压(LVSP)、收缩期左心室内压上升最大变化速率(+dp/dt_(max))和舒张期左心室内压下降最大变化速率(-dp/dt_(max))(P<0.05,P<0.01);降低血清中脑钠肽(BNP)、心肌肌钙蛋白I(cTnI)含量(P<0.05);心肌纤维排列相对紧密,胞核清晰可见,但有一定断裂,其中有少量空泡存在,未发现炎性细胞浸润,各指标有明显差异。生附子组各指标有一定程度的改善,但差异无统计学意义(P>0.05)。结论:生附子经炮制后,毒性降低,起到减毒的作用;自制黑顺片较生附子强心作用有显著增强。

基于UPLC特征图谱和含量测定的蜜远志标准汤剂质量评价研究

目的建立蜜远志标准汤剂特征图谱以及3种成分质量分数同时测定的UPLC方法,对不同来源蜜远志饮片制成的标准汤剂质量进行评价。方法采用UPLC法建立蜜远志标准汤剂的特征图谱以及细叶远志皂苷、远志酮Ⅲ和3,6′-二芥子酰基蔗糖质量分数测定的方法,并对16批标准汤剂进行测定,采用高分辨质谱对共有峰进行了指认,采用聚类分析及偏最小二乘法分析(PLS-DA)对共有峰进行了分析。结果蜜远志标准汤剂特征图谱标定了9个共有峰,通过高分辨质谱分别指认为:西伯利亚远志糖A5、西伯利亚远志糖A6、远志酮Ⅲ、3,6′-二芥子酰基蔗糖、黄花远志素A、细叶远志皂苷A、远志寡精H、远志寡糖A、1,2,3,7-四甲氧基呫吨酮,相似度结果显示16批样品的相似度均大于0.900。聚类分析及PLS-DA显示不同来源之间的样品可能存在质量差异。结论本研究建立的蜜远志标准汤剂特征图谱和质量分数测定方法简便、快速、准确,可为蜜远志相关制剂的质量评价提供参考。

熟地黄-山茱萸药对通过PI3K/AKT通路对Aβ25-35诱导的N2a细胞凋亡的影响

目的研究熟地黄-山茱萸药对通过磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/AKT)信号通路对β-淀粉样蛋白(Aβ25-35)诱导小鼠脑神经瘤细胞株(N2a)细胞凋亡的抑制作用。方法以Aβ25-35(20μmol/L)处理N2a细胞构建阿尔茨海默病(Alzheimer′s disease,AD)细胞模型,给予低、高浓度的熟地黄-山茱萸醇提物作用24 h。采用CCK8法测定细胞存活率,寻求最佳的Aβ25-35的诱导浓度;Hoechst 33342和Annexin V-FITC/PI染色检测细胞凋亡;比色法检测含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)活性;Western Blot法检测cleaved Caspase-3、p-PI3K、PI3K、p-AKT和AKT蛋白表达。结果与空白对照组比较,经过Aβ25-35处理后的N2a细胞凋亡率和Caspase-3蛋白表达及活性显著升高,p-PI3K/PI3K、p-AKT/AKT比值下降(P<0.05);给予低、高浓度的熟地黄-山茱萸醇提物作用24 h后,N2a细胞的凋亡率和Caspase-3蛋白表达及活性降低,p-PI3K/PI3K、p-AKT/AKT比值升高,与模型组比较差异有统计学意义(P<0.05);与低剂量组比,高剂量组N2a细胞的凋亡率和Caspase-3活性降低,p-AKT/AKT比值升高(P<0.05)。结论熟地黄-山茱萸醇提物可对Aβ25-35诱导的N2a细胞凋亡具有保护作用,其机制可能与促进PI3K/AKT信号通路有关。

扶阳学派运用附子之经验

基于“阳主阴从”观,扶阳学派集百年之经验,将大剂量附子灵活应用于各类急危重症和疑难杂症,取得了显著的临床疗效。文章通过阐述四川江油道地药材附子的辨证运用要点、配伍方法、剂量把握等方面的内容,并结合临床医案,对扶阳学派运用附子的用药经验进行了回顾性总结,旨在为其临床推广和应用提供可借鉴的依据。

熟地黄中促神经干细胞增殖活性的成分研究

目的 探究熟地黄中促进体外培养的神经干细胞(NSCs)增殖的活性成分。方法 采用系统溶剂萃取法及柱色谱法分离熟地黄各部位,CCK-8法测定熟地黄不同化学分离部位及活性部位所获化合物对NSCs增殖活性的影响。结果 熟地黄水提醇沉后水部位经大孔树脂、硅胶、聚酰胺分离后所得FrC5部分具有显著的促进NSCs增殖的活性(P<0.05)。FrC5部分按照溶解性不同分为FrC5-1和FrC5-2,FrC5-1部位相较于其他组别作用较强且与对照组相比差异有统计学意义(P<0.05),与对照组相比对NSCs的增殖率为75.85%。进一步分离得到2个化合物,根据MS、NMR数据及参考文献鉴定得出活性化合物为毛蕊花糖苷和异毛蕊花糖苷。毛蕊花糖苷对NSCs的增殖率为65.04%,异毛蕊花糖苷对NSCs的增殖率为35.86%。结论 毛蕊花糖苷和异毛蕊花糖苷可能是熟地黄中促进NSCs增殖的主要活性成分。

炒白芍-炙甘草配伍前后化学成分与抗炎作用相关性研究

目的明确炒白芍-炙甘草配伍对化学成分的影响及其与抗炎作用的相关性。方法采用HPLCDAD建立同时测定炒白芍-炙甘草中芍药苷、芍药内酯苷、苯甲酰芍药苷、氧化芍药苷、甘草酸、甘草苷多成分含量的分析方法;建立佐剂性关节炎大鼠模型,给药组分别灌胃炒白芍、炙甘草、炒白芍-炙甘草药液2周,ELISA检测大鼠血清炎症因子白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、前列腺素E_(2)(PGE_(2))含量;采用析因设计、双变量相关分析、多元线性回归方法研究配伍对化学成分的影响及其与抗炎药效的相关性。结果建立HPLC-DAD分析方法,炒白芍-炙甘草配伍后氧化芍药苷、芍药内酯苷、苯甲酰芍药苷含量增加(P<0.05);与模型组比较,各给药组大鼠血清IL-1β、TNF-α、PGE_(2)含量降低(P<0.01),且配伍组血清IL-1β、TNF-α、PGE_(2)含量低于炒白芍组、炙甘草组(P<0.05,P<0.01);析因设计结果显示炒白芍-炙甘草配伍存在交互效应,双变量相关分析结果显示氧化芍药苷、芍药内酯苷、芍药苷、甘草苷、苯甲酰芍药苷、甘草酸含量分别与IL-1β、TNF-α、PGE_(2)含量呈负相关,多元线性回归结果显示甘草苷、甘草酸与IL-1β、TNF-α、PGE_(2)含量呈负相关。结论本研究建立的炒白芍-炙甘草多成分含量分析方法简便快捷、专属性强、准确可靠,可用于其质量评价与控制。炒白芍-炙甘草配伍可抑制炎症因子IL-1β、TNF-α、PGE_(2)表达,且化学成分含量与抗炎作用具有相关性。

石菖蒲-熟地黄对认知功能障碍模型大鼠的网络药理学作用机制及实验研究

目的:探究石菖蒲-熟地黄对认知功能障碍模型大鼠学习记忆力的影响及作用机制,并进行实验验证。方法:在中药系统药理学数据库与分析平台(TCMSP)、人类基因数据库(GeneCards)、在线人类孟德尔遗传数据库(OMIM)中获取石菖蒲-熟地黄药对治疗认知功能障碍的潜在作用靶点,并构建蛋白质-蛋白质相互作用(PPI)网络,在生物学信息注释库(DAVID)分析平台中对共同靶点进行基因本体(GO)和京都基因和基因组百科全书(KEGG)通路富集分析。基于网络药理学预测结果,用动物实验进行验证。采用双侧脑室注射脂多糖(LPS)的方法模拟中枢神经炎症损伤引起的认知障碍模型,连续灌胃给药8周,假手术组予以注射等量生理盐水。Morris水迷宫观察各组大鼠空间学习记忆力的变化情况;酶联免疫吸附试验(ELISA)检测大鼠血清肿瘤坏死因子-α(TNF-α)含量;蛋白质印迹法(Western Blotting)检测Toll样受体4/核因子κB(TLR4/NF-κB)通路相关蛋白含量。结果:从石菖蒲-熟地黄药对中筛选到26个有效成分,疾病相关靶点64个,涉及晚期糖基化终末产物-晚期糖基化终末产物受体(AGE-RAGE)信号通路、TNF信号通路、白细胞介素17(IL-17)信号通路、Toll样受体通路、NF-κB信号通路等5条主要信号通路。动物实验结果显示,石菖蒲-熟地黄中、高剂量组大鼠逃避潜伏期明显缩短(P<0.05),在目标象限停留时间明显延长(P<0.05),TNF-α的表达水平明显降低(P<0.05);石菖蒲-熟地黄低、中、高剂量组能显著降低TLR4、p-NF-κB p65/NF-κB p65、IL-6的表达(P<0.05)。结论:石菖蒲-熟地黄可能通过TLR4/NF-κB通路减少炎症介质释放,抑制神经炎症来干预认知功能障碍的发生发展,为该药对抗认知功能障碍有效成分的开发和运用提供了方向。