Chrysophanol inhibits the progression of gastric cancer by activating nod-like receptor protein-3

Aim:Gastric cancer(GC)is one of the most common malignant tumors.Chrysophanol has been reported to possess antitumor effects on a variety of cancers;however,its role in GC remains unclear.This study aimed to investigate the effects of chrysophanol on the proliferation,pyroptosis,migration,and invasion of GC cells.Methods:Human GC cell lines MKN 28 and AGS cells were treated with different concentrations of chrysophanol,then cell proliferation,migration,invasion and pyroptosis were determined by CCK-8,colony-forming assay,wound healing assay,Transwell assay,and flow cytometry.Cell migration and invasion were reassessed in these transfected cells following the transfection of nod-like receptor protein-3(NLRP3)siRNA in MKN 28 and AGS cells.To examine the downstream signaling pathway of the NLRP3 signaling pathway,NLRP3,caspase-1,gasdermin-D,interleukin(IL)-1β,and IL-18 were detected by quantitative real-time-polymerase chain reaction or western blotting.Results:Chrysophanol inhibited the proliferation of GC cells,caused pyroptosis,inhibited cell migration and invasion,and increased the expression of NLRP3 inflammasomes in GC cells.Knockdown of NLRP3 inhibited the effects of chrysophanol on proliferation,pyroptosis,migration,and invasion of GC cells.Chrysophanol plays an anticancer role by enhancing NLRP3.Conclusions:Chrysophanol exerts anti-neoplastic effects in vitro in GC cells by modulating NLRP3,thus highlighting its therapeutic potential in GC.

Neuroprotective Effect of Chrysophanol as a PI3K/AKT/mTOR Signaling Inhibitor in an Experimental Model of Autologous Blood-induced Intracerebral Hemorrhage

Objective Intracerebral hemorrhage(ICH)refers to predominant,sporadic,and non-traumatic bleeding in the brain parenchyma.The PI3K/AKT/mTOR signaling pathway is an important signal transduction pathway regulated by enzyme-linked receptors and has many biological functions in mammals.It plays a key role in neuronal metabolism,gene expression regulation,and tissue homeostasis in the healthy and diseased brain.Methods In the present study,the role of the PI3K/AKT/mTOR pathway inhibitor chrysophanol(CPH)(10 mg/kg and 20 mg/kg,orally)in the improvement of ICH-associated neurological defects in rats was investigated.Autologous blood(20µL/5 min/unilateral/intracerebroventricular)mimics ICH-like defects involving cellular and molecular dysfunction and neurotransmitter imbalance.The current study also included various behavioral assessments to examine cognition,memory,and motor and neuromuscular coordination.The protein expression levels of PI3K,AKT,and mTOR as well as myelin basic protein and apoptotic markers,such as Bax,Bcl-2,and caspase-3,were examined using ELISA kits.Furthermore,the levels of various neuroinflammatory cytokines and oxidative stress markers were assessed.Additionally,the neurological severity score,brain water content,gross brain pathology,and hematoma size were used to indicate neurological function and brain edema.Results CPH was found to be neuroprotective by restoring neurobehavioral alterations and significantly reducing the elevated PI3K,AKT,and mTOR protein levels,and modulating the apoptotic markers such as Bax,Bcl-2,and caspase-3 in rat brain homogenate.CPH substantially reduced the inflammatory cytokines like interleukin(IL)-1β,IL-6,and tumor necrosis factor-α.CPH administration restored the neurotransmitters GABA,glutamate,acetylcholine,dopamine,and various oxidative stress markers.Conclusion Our results show that CPH may be a promising therapeutic approach for overcoming neuronal damage caused by the overexpression of the PI3K/AKT/mTOR signaling pathway in ICH-induced neurological dysfunctions in rats.

Chrysophanol attenuates lead exposure-induced injury to hippocampal neurons in neonatal mice

Previous studies have shown that chrysophanol protects against learning and memory impairments in lead-exposed adult mice. In the present study, we investigated whether chrysophanol can alleviate learning and memory dysfunction and hippocampal neuronal injury in lead-exposed neonatal mice. At the end of lactation, chrysophanol（0.1, 1.0, 10.0 mg/kg） was administered to the neonatal mice by intraperitoneal injection for 15 days. Chrysophanol significantly alleviated injury to hippocampal neurons and improved learning and memory abilities in the lead-poisoned neonatal mice. Chrysophanol also significantly decreased lead content in blood, brain, heart, spleen, liver and kidney in the lead-exposed neonatal mice. The levels of malondialdehyde in the brain, liver and kidney were significantly reduced, and superoxide dismutase and glutathione peroxidase activities were significantly increased after chrysophanol treatment. Collectively, these findings indicate that chrysophanol can significantly reduce damage to hippocampal neurons in lead-exposed neonatal mice.

Ultrastructural Changes of Sphaerotheca fuliginea (Schlechtend.:Fr.) Pollacci in Cucumber After Treated by Chrysophanol

Chrysophanol （1,8-dihydroxy-3-methylanthraquinone） is a free anthraquinone compound and a secondary metabolite of medicinal plant rhubarb. Chrysophanol has been reported to have both protective and curative activity against Sphaerotheca fuliginea （Schlechtend.：Fr.） Pollacci, the causal agent of cucumber powdery mildew. In this paper the ultrastructure of powdery mildew on cucumber leaves was studied using electron microscopy after the leaves were treated with chrysophanol. Results showed that preventive treatments with chrysophanol affected fungal development, including spore germination, appressorial formation, and penetration. In the curative treatment, chrysophanol affected fungal survival, resulting in broken cell wall of germ tubes, swelling and collapse of hyphal tips, hyphal malformation, delayed and reduced sporulation. The morphological changes induced by chrysophanol at the ultrastructural level were reflected by haustorium deformation, vacuolization, abortion, and necrosis. Host cell walls infected or adjacent to haustoria were thickened. All these morphological changes of S. fuliginea further confirmed the fungicidal activity of chrysophanol on powdery mildew of cucumber.

Advanced Oxidation Protein Products Regulate the Pharmacokinetics of Aloe-emodin,Emodin,Rhein,and Chrysophanol in Chronic Kidney Disease Rats

Background:As accelerators and products of the progression of chronic kidney disease(CKD),advanced oxidation protein products(AOPPs)affect the function of the liver.Huang Gan granules(HGGs)are commonly used to prevent the progression of CKD,but the pharmacokinetics of aloe-emodin,emodin,rhein,and chrysophanol in HGGs in CKD remain unknown.Objective:To investigate the influence and its molecular mechanism of AOPPs on the in vivo pharmacokinetics of aloe-emodin,emodin,rhein,and chrysophanol in HGGs.Methods:We constructed 5/6 nephrectomised(5/6 nx),adenine-induced(adenine)and AOPP-treated rat models.After oral administration of HGG,the concentrations of aloe-emodin,emodin,rhein,and chrysophanol in the plasma samples were detected by high-performance liquid chromatography(HPLC),and their pharmacokinetics were analysed with the PKSolver software.The plasma concentrations of IL-6 and TNF-αare detected by enzyme linked immunosorbent assay(ELISA).The RT-PCR was performed in the HepG2 cells to explore the effect of TNF-αand IL-6 on the mRNA expression of CYP1A2 and CYP3A4.Result:The results showed that the method was suitable for the quantification of four anthraquinones in plasma and excreta samples with satisfactory linear(R R^(2)>0.9931),precision(<9.4%)and accuracy(±10%).In 5/6 nx,adenine and AOPPs-treated rats,the concentrations of TNF-αand IL-6 were increased.In 5/6 nx and adenine rats,the pharmacokinetic parameters(t_(1/2),MRT_(0-∞)and AUC_(0-∞))of aloe-emodin,emodin,rhein,and chryso-phanol were,respectively,significantly increased and correlated with the concentration of AOPPs.In AOPPs-treated rats,the concentration of AOPPs was significantly increased and the pharmacokinetic parameters of four anthraquinones were also increased.Conclusion:In summary,inflammatory cytokine production may be one of the important causes in AOPPs’regulat-ing the pharmacokinetic of aloe-emodin,emodin,rhein,and chrysophanol in the CKD rats.Studies of aloe-emodin,emodin,rhein,and chrysophanol in CKD facilitate the appropriate prescription of HGGs in the clinical.

HPLC法测定十三味逐瘀合剂中游离大黄酚含量

目的建立十三味逐瘀合剂中大黄酚高效液相含量测定的方法。方法采用高效液相色谱法(HPLC法)对十三味逐瘀合剂中游离大黄酚含量进行测定,从提取溶剂、取样量、提取时间等影响因素中选出最优条件,建立完整的含量测定方法。结果用thermos C18色谱柱(250 mm×4.6 mm,5μm);以甲醇-0.1%磷酸(75∶25,V/V)为流动相,采用等度洗脱法,254 nm波长的色谱条件;以甲醇为浸膏提取溶剂,超声30 min的提取方法;大黄酚质量浓度在1.63~16.32μg/mL(R^(2)=0.9995)范围内具有良好的线性关系;平均回收率为104.17%;测得十三味逐瘀合剂中游离大黄酚平均含量为15.32μg/mL。结论本实验所建立的方法操作简单,专属性强,重现性好,可用于十三味逐瘀合剂中游离大黄酚的含量测定。

Chrysophanol protects against doxorubicin-induced cardiotoxicity by suppressing cellular PARylation

The clinical application of doxorubicin(DOX) in cancer chemotherapy is limited by its lifethreatening cardiotoxic effects. Chrysophanol(CHR), an anthraquinone compound isolated from the rhizome of Rheum palmatum L., is considered to play a broad role in a variety of biological processes.However, the effects of CHR’s cardioprotection in DOX-induced cardiomyopathy is poorly understood. In this study, we found that the cardiac apoptosis, mitochondrial injury and cellular PARylation levels were significantly increased in H9 C2 cells treated by Dox, while these effects were suppressed by CHR. Similar results were observed when PARP1 activity was suppressed by its inhibitors 3-aminobenzamide(3 AB)and ABT888. Ectopic expression of PARP1 effectively blocked this CHR’s cardioprotection against DOX-induced cardiomyocyte injury in H9 C2 cells. Furthermore, pre-administration with both CHR and 3 AB relieved DOX-induced cardiac apoptosis, mitochondrial impairment and heart dysfunction in Sprague–Dawley rat model. These results revealed that CHR protects against DOX-induced cardiotoxicity by suppressing cellular PARylation and provided critical evidence that PARylation may be a novel target for DOX-induced cardiomyopathy.

Photogeneration of the free radicals and singlet oxygen by chrysophanol from rheum

In the current work, chryosphanol (MHAQ) was isolated and purified from rheum, and the photosensitization activities were studied. In nitrogen-saturated DMSO solution, irradia- tion of chryosphanol (MHAQ) with visible light (>430 nm) produced the semiquinone radical anions (MHAQ??), which was intensified significantly by an electron donor NADH, suggesting electron transfer between the excited and ground state photosensitizer molecules. In an air-saturated DMSO solution, superoxide radical anions (O2 ), trapped by DMPO, were detected. ?? It was proved that O2 was not originated from the singlet oxygen (1O2) but dependent on DMPO, ?? chrysophanol, oxygen and the irradiation time. The singlet oxygen and OH could also be pro- ? duced by the photosensitization of the photosensitizer. These suggest that chrysophanol pos- sesses the photosensitization activity via Type I mechanism and Type II mechanism. To evaluate the photosensitization activities of MHAQ, emodin and mixed anthraquinone derivatives ex- tracted from rheum, the relative productivities of O2 were estimated to be 1.8, 1.1 and 1.0 re- ?? spectively and the quantum yields for singlet oxygen to be 0.36, 0.53 and 0.14 respectively. Therefore, these low-cost pigments may be potentially used as phototherapeutic medicine for some kind of vascular diseases or photo-activated pesticides.

Chrysophanol localizes in mitochondria to promote cell death through upregulation of mitochondrial cyclophilin D in HepG2 cells

Objective:Chrysophanol(Chry) displays potent anticancer activity in human cancer cells and animal models,but the cellular targets of Chry have not been fully defined.Herein,we speculated whether mitochondria were a target involved in Chry-induced cytotoxicity.Methods:Human liver cancer cell line HepG2 was incubated.The cytotoxicity was evaluated by MTT assay.Mitochondria localization was evaluated by a confocal microscopy.Mitochondrial membrane potential ΔΨm was detected by TMRE staining and determined by the flow cytometer.The levels of ATP,mitochondrial superoxide anions,and GSH/GSSG were determined according to the assay kits.The apoptosis were evaluated through Hoechst33342/PI and Annexin V/PI staining,respectively.The expression of cyclophilin D(CyPD) was determined by immunoblot method,and the interaction between CyPD and Chry was analyzed by molecule docking procedure.Results:Chry itself mainly localized in mitocho ndria to cause mitochondrial dysfunction and cell death in HepG2 cells.As regard to the mechanism,cyclosporin A as the inhibitor for the formation of mitochondrial permeability transition pore(mPTP) moderately suppressed cell death,indicating mPTP involved in the process of cell death.Further,Chry enhanced the protein expression of Cyclophilin D(CyPD) which is a molecular componentry and a modulator of mPTP,while antioxidant N-acetyl-L-cysteine inhibited the expression of CyPD.Molecule docking procedure disclosed two hydrogen-bonds existed in CyPD-Chry complex with-11.94 kal/mol of the binding affinity value.Besides,the mtDNA-deficient HepG_2-ρ0 cells were much resistant to Chry-induced cell death,indicating mtDNA at least partly participated in cell death.A combination of Chry and VP-16 produced the synergism effect toward cell viability andΔΨm,while Chry combined with Cis-Pt elicited the antagonism effect.Conclusion:Taken together,enrichment in mitochondria and actions on mPTP,CyPD and mtDNA provides an insight into the anticancer mechanism of Chry.The combination therapy for Chry with clinical drugs may deserve to further explore.

HPLC法测定骨伤跌打康复凝胶贴膏剂中7种成分含量

目的 建立测定骨伤跌打康复凝胶贴膏剂中7种主要成分质量分数的HPLC法。方法 色谱柱为Agilent ZORBA×SB-C18S tableBond Analytical柱(4.6 mm×250 mm, 5μm);测定波长为290 nm,流动相为乙腈-0.1%甲酸水梯度洗脱;流速为1.0 mL/min;柱温为35℃;进样量为10μL。结果 骨伤跌打康复凝胶贴膏剂中的原儿茶酸、咖啡酸、阿魏酸、盐酸小檗碱、大黄酸、大黄素、大黄酚7种主要有效成分在浓度线性范围内与色谱峰面积的标准曲线线性关系良好;原儿茶酸、咖啡酸、阿魏酸、盐酸小檗碱、大黄酸、大黄素、大黄酚的平均回收率分别为93.79%、95.86%、96.12%、95.31%、96.58%、107.43%、97.12%,7种成分的平均回收率均在93.79%~107.43%范围内。结论 建立的方法适合作为骨伤跌打康复凝胶贴膏剂质量控制的方法。

生肌化瘀凝胶质量标准研究

目的建立生肌化瘀凝胶的质量控制方法。方法采用薄层色谱法对样品中的血竭、大黄、白芷、丹参、紫草、黄芪进行定性鉴别;采用高效液相色谱法测定样品中血竭素和大黄酚的含量,血竭素和大黄酚的色谱柱分别为Agilent Zorbax-SB-C18柱(250 mm×4.6 mm,15μm)和Zorbax Eclipse XDB-C18柱(250 mm×4.6 mm,5μm),流动相分别为乙腈-0.05 mol/L磷酸二氢钠溶液(50∶50,V/V)和甲醇-0.1%磷酸水溶液(85∶15,V/V),流速为1 mL/min,检测波长分别为440 nm和254 nm,柱温分别为40℃和30℃,进样量为10μL。结果薄层色谱图中特征斑点显色清晰,分离度好,且阴性对照无干扰。血竭素、大黄酚的质量浓度分别在0.02585~0.09048 mg/mL和0.001~0.060 mg/mL范围内与峰面积线性关系良好(r=0.9999,n=6);精密度、稳定性、重复性试验结果的RSD均低于3.0%;平均加样回收率分别为101.88%和96.82%,RSD分别为2.50%和0.79%(n=9)。6批样品中,血竭素、大黄酚的平均含量分别为0.0396 mg/mL和0.0037 mg/mL。结论该方法专属性强、结果准确,可用于生肌化瘀凝胶的质量控制。

HPLC法同时测定肾衰宁胶囊中5种大黄游离蒽醌的含量

目的:建立HPLC法同时测定肾衰宁胶囊中芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚的含量。方法:采用资生堂MGⅡC_(18)色谱柱(5μm,4.6 mm×250 mm),流动相为甲醇(A)-0.1%磷酸水(B),二元梯度洗脱,检测波长为254 nm,柱温30℃,流速为1.0 mL·min^(-1),进样量10μL。结果:芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚的进样量范围分别在0.008614~2.1534μg(r=0.9999)、0.006823~1.7058μg(r=0.9999)、0.008799~1.7598μg(r=0.9999)、0.03771~9.428μg(r=0.9999)和0.003623~0.9058μg(r=0.9999),与峰面积呈良好的线性关系;平均回收率(n=9)分别为97.42%(RSD=0.8%)、103.58%(RSD=1.1%)、101.45%(RSD=0.8%)、103.54%(RSD=0.6%)、103.35%(RSD=1.1%)。结论:建立的方法准确、简便、灵敏,可用于肾衰宁胶囊中芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚的含量测定,可为肾衰宁胶囊的质量控制研究提供有益参考。

The mechanism of Chuanxiong Rhizoma on glaucomatous optic nerve injury based on network pharmacology and molecular docking

Based on network pharmacology,this study predicted the potential molecular mechanism and related pathways of the protective effect of traditional Chuanxiong Rhizoma,a traditional Chinese herb,on glaucomatous optic nerve injury,and conducted in vitro experimental verification of the predicted results of network analysis.We analyzed the molecular mechanism of Chuanxiong Rhizoma in the potential treatment of glaucoma by revealing its main active ingredients and predicting its targets,so as to provide reference for subsequent basic research.Network pharmacological research results showed that the potential hub targets and key signaling pathways of Chuanxiong Rhizoma in the treatment of glaucoma were closely related to biological processes such as apoptosis,autophagy,inflammation,oxidative stress and angiogenesis.Molecular docking showed that many active ingredients,such as chrysophanol(CHR),myricanone and retinol,could combine well with their target proteins by intermolecular forces,especially CHR had strong binding ability with each target.We speculated that the main active component of Chuanxiong Rhizoma might be involved in the regulation of PI3K-Akt,Nod-like receptor,IL-4 and IL-13,MAPK,AGE-RAGE and neurotrophin signaling pathway by regulating of PI3K,Akt,TLR4,RAGE,NTRK2 and other key targets.Furthermore,it may achieve multi-directional intervention on apoptosis/autophagy,inflammation/immunity,oxidative stress and nutrient metabolism of axoplasma flow,and then delay the degeneration of optic nerve injury.In vitro experiments showed that the active component CHR of Chuanxiong Rhizoma could reverse the M1-type polarization and autophagy/apoptosis of mouse microglia(BV2)induced by lipopolysaccharide(LPS)at the transcriptional level.Meanwhile,the expression of inflammatory mediators IL-1βand TNF-αwas inhibited,and the mRNA level of anti-inflammatory factor IL-10 was significantly increased.In addition,CHR down-regulates activation of the RAGE-NOX4 pathway mediated by LPS in reducing oxidative stress.In this study,network pharmacology and molecular docking technology were integrated for the first time to explore the potential molecular mechanism of traditional Chinese herb“Chuanxiong Rhizoma”in the treatment on glaucoma,and CHR was innovatively proposed as an important ingredient in Chuanxiong Rhizoma that plays a protective role in the damage of optic nerve.Preliminary verification was conducted through in vitro experiments.The results suggest that Chuanxiong Rhizoma may interfere with autophagy and apoptosis,inhibit immune inflammation,as well as reduce oxidative stress in the treatment of glaucoma through the active components represented by CHR,so as to resist progressive optic nerve injury.Our study provides theoretical basis for the clinical use of Chinese herbal medicine or its extract in glaucoma,and also lays a solid foundation for the research of Chinese medicine in the field of optic nerve protection.

大黄提取物中五种蒽醌类活性成分的同步检测方法

目的:建立高效液相色谱-紫外分析法(HPLC-UV)同步检测大黄提取物中五种蒽醌类活性成分。方法:优化蒽醌类活性成分同步检测方法的溶剂、色谱分离条件,并从标准曲线线性、精密度、准确度、回收率、稳定性及重现性等方面对方法进行了验证,采用验证的方法检测了大黄提取物中五种蒽醌类成分的含量。结果:分别采用氯仿、二甲基亚砜、甲醇作为不同对照品储备液溶剂,采用甲醇∶水(50∶50,V∶V)作为对照品工作溶液溶剂,采用甲醇配制样品溶液。最佳色谱分离条件:采用Symmetry C_(18)(5μm,4.6 mm×250 mm)色谱柱,采用二元泵等度洗脱方式,流动相A为含0.2%甲酸的水溶液,流动相B为乙腈(A∶B体积比为15∶85),流速1.30mL/min,柱温30℃,检测波长254 nm,进样体积20.0μL。方法曲线拟合度好,R2均大于0.9999;6份平行样品芦荟大黄素、大黄素、大黄酸、大黄酚和大黄素甲醚的RSD分别为1.10%、1.05%、0.74%、0.84%和0.82%,回收率为94.54%~98.70%;室温放置22 h以及4℃条件下储存20 d基本稳定,自动进样器温度下存储24 h重新进样重现性良好。采用建立的方法测得大黄冻干粉样品中芦荟大黄素、大黄素、大黄酸、大黄酚和大黄素甲醚的含量分别为0.900、0.639、3.506、0.780、0.304 mg/g。结论:本研究建立了可同步检测芦荟大黄素、大黄素、大黄酸、大黄酚和大黄素甲醚等五种蒽醌类活性成分的高效液相色谱-紫外分析法(HPLC-UV),经验证其具有良好的线性关系、准确性、精密度、稳定性和重现性。

CUMS抑郁小鼠模型海马氧化损伤的研究及大黄酚的保护作用

目的:探讨大黄酚(Chrysophanol,CHR)对于慢性不可预见性温和刺激(CUMS)导致的抑郁小鼠氧化应激的影响。方法:采用CUMS方法建立小鼠抑郁模型,分析不同剂量大黄酚对小鼠抑郁样行为的变化和小鼠海马组织超氧化物歧化酶(SOD)和丙二醛(MDA)水平的影响。结果:CHR能显著增加抑郁模型小鼠的体重,提高糖水偏好百分比,显著降低小鼠海马体中MDA水平,显著升高小鼠海马体SOD水平。结论:大黄酚能有效改善CUMS大鼠的抑郁样行为,其机制可能与氧化应激的调控有关。

大黄酚对CUMS抑郁小鼠行为学的改善作用

目的探讨大黄酚对慢性不可预知性应激刺激(CUMS)导致的小鼠的抑郁行为学的改善作用。方法将40只BALB/c小鼠分为对照组、模型组及大黄酚低(0.1 mg/kg)、中(1 mg/kg)、高剂量组(10 mg/kg),每组各8只。对模型组及大黄酚低、中、高剂量组给予CUMS建立小鼠抑郁模型,给药均采用腹腔注射方法。通过比较各组小鼠体重、糖水偏好度和抑郁样行为变化,测试大黄酚改善抑郁模型的效果。结果实验第21天,与对照组比较,模型组及大黄酚组体重和糖水偏好度显著降低(P<0.05),提示小鼠抑郁模型建立成功。实验第42天,与模型组相比,大黄酚低、中、高剂量组体重和糖水偏好度均显著上升(P<0.05),悬尾不动时间显著缩短(P<0.05),大黄酚中、高剂量组强迫游泳不动时间明显缩短(P<0.05)。结论大黄酚能有效改善CUMS大鼠的抑郁样行为。

大黄酚通过核因子E2相关因子2/血红素氧合酶-1信号通路对糖尿病足溃疡大鼠新生血管生成及氧化应激的影响

目的:探讨大黄酚通过核因子E2相关因子2(Nrf2)/血红素氧合酶-1(HO-1)信号通路对糖尿病足溃疡大鼠新生血管生成及氧化应激的影响。方法:构建糖尿病足溃疡大鼠模型。将48只建模成功大鼠随机分为模型组、大黄酚组(给予30 mg/kg)、ML385组(Nrf2抑制剂,给予30 mg/kg)、大黄酚+ML385组(给予30 mg/kg大黄酚+30 mg/kg ML385),每组12只。另取12只大鼠作为对照组。各组给予相应药物干预4周。末次给药结束后24 h,测定创面愈合率,酶联免疫吸附法(ELISA)检测血清血管内皮生长因子(VEGF)、肿瘤坏死因子-α(TNF-α)以及创面组织碱性成纤维细胞生长因子(bFGF)、血管性血友病因子(vWF)水平;HE染色观察创面组织病理变化;检测血清中丙二醛(MDA)、超氧化物歧化酶(SOD)水平;蛋白免疫印迹法检测创面组织Nrf2、HO-1蛋白表达。结果:对照组创面组织中炎性细胞较少,成纤维细胞较多,可见丰富的毛细血管。与对照组比较,模型组创面组织中可见大量炎性细胞,新生毛细血管分布较少,创面愈合率、血清VEGF以及创面组织bFGF、vWF、SOD水平和Nrf2、HO-1蛋白表达降低,血清TNF-α、创面组织MDA水平升高(均P<0.05)。与模型组比较,大黄酚组创面组织中可见大量新生毛细血管,少量炎性细胞,创面愈合率、血清VEGF以及创面组织bFGF、vWF、SOD水平和Nrf2、HO-1蛋白表达升高,血清TNF-α、创面组织MDA水平降低(均P<0.05);ML385组创面组织中新生毛细血管显著减少,炎性细胞显著增多,创面愈合率、血清VEGF以及创面组织bFGF、vWF、SOD水平和Nrf2、HO-1蛋白表达降低,血清TNF-α、创面组织MDA水平升高(均P<0.05)。大黄酚+ML385组大鼠创面组织病理变化和上述指标介于大黄酚组与ML385组之间(均P<0.05)。结论:大黄酚能促进糖尿病足溃疡大鼠新生血管的生成,抑制炎症反应和氧化应激,其机制可能与激活Nrf2/HO-1信号通路有关。

UPLC双波长切换法同时测定开胸消食片中6个成分的含量

目的建立UPLC双波长切换法同时测定开胸消食片中的甘草苷、橙皮苷、厚朴酚、和厚朴酚、大黄素和大黄酚的含量。方法采用Wa⁃ters公司的Acquity UPLCR BEH C_(18)柱(150×2.1 mm,1.7μm);流动相为乙腈⁃0.1%磷酸,梯度洗脱;流速0.2 mL·min^(-1),柱温30℃,检测波长280 nm(甘草苷与橙皮苷)和255 nm(厚朴酚、和厚朴酚、大黄素与大黄酚)。结果甘草苷、橙皮苷、厚朴酚、和厚朴酚、大黄素、大黄酚的线性范围分别为0.41~20.71μg·mL^(-1)、5.15~257.31μg·mL^(-1)、2.46~122.96μg·mL^(-1)、0.86~43.11μg·mL^(-1)、0.48~24.00μg·mL^(-1)、3.93~196.64μg·mL^(-1)(r≥0.9995)。6个成分的平均回收率在92.9%~104.7%之间。结论本实验建立的方法操作方便、准确可靠、重复性好并且分离效果好,可用于开胸消食片中6个成分的含量测定。

HPLC法同时测定中消软膏中4种成分含量

目的:建立HPLC法同时测定中消软膏中盐酸小檗碱、芦荟大黄素、大黄酚、欧前胡素4种活性成分的含量。方法:采用超声提取法,提取溶剂为甲醇,流动相为乙腈(A)-0.1%的磷酸水溶液(B),检测波长为254 nm,体积流量0.8 mL/min,柱温30℃,理论塔板数以盐酸小檗碱计算不低于4000。结果:中消软膏中盐酸小檗碱、芦荟大黄素、大黄酚、欧前胡素在线性范围内线性关系良好,平均回收率分别为99.6%、99.8%、99.1%、99.7%,RSD分别为1.0%、0.8%、1.1%、0.3%。结论:该方法简便、快速、精密度和准确度良好,可为中消软膏质量标准提升提供一定依据。

怀牛膝中化合物的分离鉴定

从怀牛膝乙醇提取物中分离出８个化合物，通过理化常数测定和光谱解析并与文献对照，分别鉴定为α－菠甾醇（１），β－谷甾醇（２），大黄酚（３），邻苯二甲酸二正丁酯（４），软脂酸（５），α－菠甾醇葡萄糖甙（６），胡萝卜甙（７）和脱皮甾酮（８），１～７为首次自该植物中分离得到。