丹酚酸B和人参皂苷Rg1合用对卒中后出血性转化的抑制作用

目的:对脑卒中患者进行血管再通治疗不仅受时间窗的限制,还伴随着出血性转化的风险。本研究在小鼠大脑中动脉阻塞(MCAO)模型上观察了丹酚酸B和人参皂苷Rg1合用(Salb/Rg1)对脑缺血时间窗和出血性转化的影响。方法:通过脑梗死体积、神经行为障碍和组织形态学检查评估SalB/Rg1的保护作用和时间窗。对MCAO小鼠进行葡萄糖刺激和再灌注考察SalB/Rg1对出血性转化的抑制。SalB/Rg1对出血性转化的抑制作用是通过免疫荧光染色和原位明胶酶谱技术来确定的。结果:通过组织病理学染色根据细胞结构进一步确定SalB/Rg1以剂量依赖的方式显著减少了梗塞体积并改善了神经行为,且SalB/Rg1对卒中的保护时间窗长达9h。其次,SalB/Rg1下调了出血评分、梗死体积并改善异常神经行为。最后,发现SalB/Rg1对出血性转化的抑制与它对神经血管单元完整性的保护相伴随。在梗死的边缘区域,SalB/Rg1减少了星形胶质细胞的激活,保持了内皮细胞之间连接蛋白(Claudin-5)的丰度,大大降低了基质金属肽酶9(MMP-9)的活性。结论:SalB/Rg1是一种很有前景的抗卒中,尤其是抗出血性转化的新策略。

Nonclinical Study of the Active Components of Doxorubicin Hydrochloride Liposome Injection in Vivo

Objectives: A non-clinical study was performed to establish a LC-MS/MS method to determine the in vivo active components of doxorubicin hydrochloride liposome injection in the plasma of Sprague-Dawley rats. Methods: Ten male SD rats were administered tail vein with a single dose of 10 mg/kg, and the concentrations of doxorubicin hydrochloride in plasma, heart, liver, spleen, lung, and kidney were determined by liquid chromatography-tandem mass spectrometry, and the pharmacokinetic parameters were calculated. Results: The final concentration of doxorubicin hydrochloride ranged from 500 ng/mL to 250,000 ng/mL, and the lower limit of quantification was 500 ng/mL;the main pharmacokinetic parameters: T1/2 was (19.282 ± 10.305) h, Cmax was (118514.828 ± 26155.134) ng/mL, AUC0-24 and AUC0-∞ were (1216659.205 ± 192706.268) ng/mL⋅h and (2082244.523 ± 860139.487) ng/mL⋅h, MRT0-24 and MRT0-∞ were (9.237 ± 0.423) h and (26.52 ± 14.015) h, respectively, and clearance (CL) was (0.005 ± 0.002) mL/h⋅ng. Conclusions: The method is simple, rapid, and sensitive, which can be used for the determination of doxorubicin hydrochloride concentration in the plasma of SD rats and pharmacokinetic non-clinical studies.

Simultaneous determination of amino acids in different teas using supercritical fluid chromatography coupled with single quadrupole mass spectrometry

Tea is a widely consumed beverage and has many important physiological properties and potential health benefits. In this study, a novel method based on supercritical fluid chromatography coupled with mass spectrometry (SFC-MS) was developed to simultaneously determine 11 amino acids in different types of tea (green teas, Oolong tea, black tea and Pu-erh tea). The separation conditions for the analysis of the selected amino acids including the column type, temperature and backpressure as well as the type of additive, were carefully optimized. The best separation of the 11 amino acids was obtained by adding water (5%, v/v) and trifluoroacetic acid (0.4%, v/v) to the organic modifier (methanol). Finally, the developed SFC-MS method was fully validated and successfully applied to the determination of these amino acids in six different tea samples. Good linearity (r ≥ 0.993), precision (RSDs≤ 2.99%), accuracy (91.95%-107.09%) as well as good sample stability were observed. The limits of detection ranged from 1.42 to 14.69 ng/mL, while the limits of quantification were between 4.53 and 47.0 ng/mL. The results indicate that the contents of the 11 amino acids in the six different tea samples are greatly influenced by the degree of fermentation. The proposed SFC-MS method shows a great potential for further investigation of tea varieties.

Synthesis, Crystal Structure and Na^+/K^+-ATPase Inhibitory Activity of △^(14,15)-anhydro-24-thiocarbonylbufalin

The title compound △14,15-anhydro-24-thiocarbonylbufalin （1） was prepared by the reaction of natural product bufalin with Lawesson reagent. The crystal structure of 1, C24H3002S＇C24H3002S, was determined by single-crystal X-ray diffraction analysis. It belongs to monoclinic, space group C2, with a = 30.9845（2）, b = 6.8036（3）, c = 22.5791（15）/k, V= 4241.7（4） A3, Mr = 384.21, Z = 4, Dc= 1.204 g/cm3,μ = 1.463 mm4, F（000） = 1664, S = 1.064, R = 0.0487 and wR = 0.0645 for 4683 unique reflections, of which 3757 were observed （I 〉 2σ（/））. The asymmetric unit contains two independent molecules （ⅠandⅡ）, which are closely similar to each other except for the orientation of the lactone ring. Both conformations of I and II are in good agreement with the solution structure in methanol as indicated by 1H-NMR analysis. Due to the presence of heavy atom sulfur in the molecules, the final refinement resulted in a small Flack parameter 0.02（3）, permitting the assignments of the absolute configuration. In the solid state, intermolecular hydrogen bonds involving thiocarbonyl group in the lactone moiety and the hydroxyl groups in the steroid moiety ester linked adjacent molecules into a three-dimensional network. Compound 1 showed weak inhibition on Na＋/K＋-ATPase in contrast to the strong inhibitory activity of the parent compound bufalin, suggesting that the carbonyl group in lactone moiety and the hydroxyl group atC-14 play important roles for the inhibition of Na＋/K＊-ATPase.

Study on the Preparation and Characterization of Glycyrrhizic Acid Liposomes

The purpose of this study was to prepare glycyrrhizic acid nanoliposomes and evaluate the encapsulation efficiency and other properties of glycyrrhizic acid nanoliposomes, which would provide a reference for further research on Glycyrrhizic Acid in the treatment of liver diseases. Firstly, the preparation conditions of glycyrrhizic acid liposomes were optimized by the orthogonal design method. Then, glycyrrhizic acid liposomes were prepared by ultrasonic-film dispersion method and the encapsulation efficiency was determined by the HPLC method. Finally, the properties of glycyrrhizic acid nanoliposomes were also studied. The results showed that the optimal preparation conditions of liposomes were as follows: the ratio of drug to lipid was 1:30;Lecithin: cholesterol = 1:1;Hydration medium: pure water;Ultrasonic time: 120 s. The encapsulation efficiency of liposomes was about 90%. The final liposomes were round and uniform in distribution, with an average particle size of about 50 nm and absolute zeta potential of −28.9 mV. In this study, glycyrrhizic acid liposomes were prepared and the optimal preparation conditions were optimized. The encapsulation efficiency of the liposomes under the optimized conditions was determined. The evaluation of the morphology, size, particle size and stability of the liposomes was completed.

Maternal helminth infection protects offspring from high-fat-diet-induced obesity through altered microbiota and SCFAs

Helminth-induced Th2 immunity and gut microbiota have been recently shown to be highly effective in modulating metabolic syndromes in animal models.This study aimed to determine whether maternal immunity and microbial factors affect the induction and development of obesity in offspring.Here,Heligomosomoides polygyrus(Hp)-infected or control female C57BL/6J mice mated with normal males and their offspring were fed a high-fat diet(HFD)for 9 weeks after weaning.Our results showed that Hp-induced maternal outcomes during gestation and lactation significantly impacted offspring metabolic phenotypes.This was evidenced by results showing that offspring from helminth-infected mothers on an HFD(Hp-offspring+HFD)gained significantly less body weight than those from uninfected mothers(Cont-offspring+HFD).Hp-offspring+HFD exhibited no Th2 phenotype but displayed a pattern of gut microbiota composition similar to that of Hp-infected mothers.Cross-fostering experiments confirmed that the helminth-induced maternal attenuation of offspring obesity was mediated through both prenatal and postnatal effects.Our results further showed that helminth-infected dams and their offspring had a markedly altered gut microbiome composition,with increased production of short-chain fatty acids(SCFAs).Intriguingly,Hp-infected mothers and Hp-offspring+HFD showed increased SCFA receptor(GPR)expression in adipose and colonic tissues compared to noninfected mothers and Cont-offspring+HFD,respectively.Moreover,SCFA supplementation to the pups of uninfected control mothers during lactation protected against HFD-induced weight gain,which corresponded with changes in gut bacterial colonization.Collectively,our findings provide new insights into the complex interaction of maternal immune status and gut microbiome,Hp infection,and the immunity and gut microbiome in obese-prone offspring in infant life.

Therapeutic Mechanism of Kai Xin San on Alzheimer's Disease Based on Network Pharmacology and Experimental Validation

Objective: To explore the specific pharmacological molecular mechanisms of Kai Xin San(KXS)on treating Alzheimer’s disease(AD) based on network pharmacology and experimental validation. Methods:The chemical compounds of KXS and their corresponding targets were screened using the Encyclopedia of Traditional Chinese Medicine(ETCM) database. AD-related target proteins were obtained from MalaCards database and DisGeNET databases. Key compounds and targets were identified from the compound-targetdisease network and protein-protein interaction(PPI) network analysis. Functional enrichment analysis predicted the potential key signaling pathways involved in the treatment of AD with KXS. The binding affinities between key ingredients and targets were further verified using molecular docking. Finally, the predicted key signaling pathway was validated experimentally. Positioning navigation and space search experiments were conducted to evaluate the cognitive improvement effect of KXS on AD rats. Western blot was used to further examine and investigate the expression of the key target proteins related to the predicted pathway. Results: In total, 38active compounds and 469 corresponding targets of KXS were screened, and 264 target proteins associated with AD were identified. The compound-target-disease and PPI networks identified key active ingredients and protein targets. Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis suggested a potential effect of KXS in the treatment of AD via the amyloid beta(Aβ)-glycogen synthase kinase-3 beta(GSK3β)-Tau pathway. Molecular docking revealed a high binding affinity between the key ingredients and targets. In vivo,KXS treatment significantly improved cognitive deficits in AD rats induced by Aβ1-42, decreased the levels of Aβ, p-GSK3β, p-Tau and cyclin-dependent kinase 5, and increased the expressions of protein phosphatase 1alpha(PP1A) and PP2A(P<0.05 or P<0.01). Conclusion: KXS exerted neuroprotective effects by regulating the Aβ-GSK3β-Tau signaling pathway, which provides novel insights into the therapeutic mechanism of KXS and a feasible pharmacological strategy for the treatment of AD.

Isolation and chemotaxonomic significance of stenine- and stemoninine-type alkaloids from the roots of Stemona tuberosa

One new stenine-type alkaloid, tuberostemonine D （1）, together with five known stenine-type alkaloids （2-6） and two known stemoninine-type alkaloids （7 and 8）, were isolated from the roots of Stemona tuberosa. Their structures were elucidated by extensive spectroscopic methods （IR, UV, MS, 1D and 2D NMR）, and the structure of I was further confirmed by X-ray diffraction analysis. First simultaneous isolation ofstenine- and stemoninine-type alkaloids not only added a new chemical type and increased the chemical diversity of Stemona tuberosa, but also provided chemotaxonomic clues for the close relationship between genera Stemona and Stichoneuron, and favored retaining them in the same family.

Synthesis of Resveratrol Analogs/Stilbene Derivatives and Their Nitric Oxide Inhibitory and Radical Scavenging Activities

Sixteen resveratrol analogs（stilbene derivatives） including resveratrol were synthesized, in which hy- droxystilbenes, naturally occurring polyphenols, showed protective effects against reactive oxygen and nitrogen spe- cies（ROS/RNS） and hexahydroxystilbene（16） displayed the strongest activities against NO and l,l-diphenyl- 2-picrylhydrazyl（DPPH） with IC5o values of 7.8 and 5.7 μmol/L, respectively.