芍药苷对大鼠心肌缺血再灌注损伤的保护作用及机制研究

目的:研究芍药苷(Paeoniflorin)预处理对大鼠心肌缺血再灌注损伤(MIRI)的保护性作用并对其可能机制进行探讨。方法 :制备大鼠心肌缺血再灌注损伤模型,随机分为模型组、假手术组、芍药苷60mg/kg组、30mg/kg组(n=20);取心脏测定心肌梗死面积,HE染色观察组织细胞结构,免疫组化检测心肌组织中NF-κB和ICAM-1的表达;取血测定血清中LDH、AST、CK和CK-MB含量。结果:与模型组比较,芍药苷60mg/kg组和30mg/kg组心肌梗死面积均明显降低;HE染色切片提示各给药组心肌细胞损伤较模型组为轻,且具有浓度相关性;芍药苷60mg/kg组、30mg/kg组NF-κB、ICAM-1的表达明显降低;芍药苷60mg/kg组血清CK含量降低,同时该组CK-MB亦降低显著。结论:芍药苷预处理对大鼠MIRI具有保护性作用,其机制与抑制细胞膜脂质过氧化,以及通过抑制NF-κB蛋白表达进而影响受NF-κB调控的下游炎性反应途径有关。

芍药苷对Lewis肺癌自发肺转移小鼠的抑瘤作用

目的:应用Lewis肺癌移植瘤小鼠模型,观察芍药苷的抑瘤及抗肿瘤转移作用。方法:C57BL/6J小鼠右腋皮下接种Lewis肺癌瘤株,隔天腹腔注射环磷酰胺,共3次,治疗组连续灌胃芍药苷24天,检测小鼠体重、瘤体大小,实验结束后处死动物,摘取瘤体称重,计算肿瘤生长抑制率。剥离鼠肺,解剖镜下计数肺部正反面及肺叶间的转移瘤集落数。结果:环磷酰胺60 mg/kg及芍药苷40 mg/kg对荷瘤小鼠肿瘤的生长均有明显的抑制作用,抑瘤率分别为57.26%和69.24%。环磷酰胺组及芍药苷组Lewis肺癌荷瘤小鼠肺转移集落明显少于模型组。结论:芍药苷能抑制Lewis肺癌荷瘤小鼠肿瘤生长,并可抑制Lewis肺癌荷瘤小鼠的肺转移。

Population pharmacokinetics of paeoniflorin in guanxin Ⅱ prescription

To evaluate the effect of components in Guanxin Ⅱ prescription on the pharmacokinetic profiles of paeoniflorin. Plasma concentration of Paeoniflorin in rats after intravenous injection of Paronia Pall Extract （PPE） and oral administration of PPE and three types of decoctions in Guanxin Ⅱ prescription, respectively, were determined by HPLC analyses. NONMEM （nonlinear mixed-effect modeling） method was used to analyze full set of pharmacokinetic data directly. A two-compartment model with first-order degradation in absorption compartment was employed for the data analysis. The mean of population parameters, CL1, V1, CL2, V2, Ka0, and Kal, were measured to be 0.509 L/h, 0.104 L, 0.113 L/h, 0.123 L, 0.135/h, and 0.0135/h, respectively. Inter-individual variabilities were estimated and dose formulation （DF） was identified as a significant covariate of Ka 1, Ka0, and V1. It is concluded that the pharmacokinetic behaviors of paeoniflorin in rats can alter with different dose formulations.

Assay of Peoniflorin in Gangbangui Capsule

[Objective] We aimed to establish a rapid determination method of peoniflorin in Gangbangui capsule by using HPLC.[Method] Sinochrom ODS-AP column （5 μm,4.6 mm× 250 mm） was used in our research.The mobile phase was methanol-0.3％ phosphoric acid （35：65）.The column temperature was 30 ℃.The flow rate was 0.8 ml/min.The detection wavelength was 230 nm.[Result] The concentration of peoniflorin in Gangbangui capsule ranged between 0.040 54 and 0.2432 μg/μl and a good linear relation between concentration and peak area was shown （r=0.999 99）.The average recovery was 98.89％ （RSD=2.17％）.[Conclusion] The method was fast,easy and reproducible and could be used as a quality indicator of the compound.

Production of paeoniflorin and albiflorin by callus tissue culture of Paeonia lactiflora Pall

In order to facilitate the preparation of paeoniflorin(PF)and albiflorin(AF),two chief bioactive constituents in Paeonia lactiflora Pall(PL),induction and culture of callus from PL were studied.With a modified woody plant medium supplemented with 0.5 mg·L-16-benzylaminopurine,1.0 mg·L-1naphthylacetic acid,0.1 mg·L-1thidiazuron and 30 g·L-1sucrose,callus was induced from four kinds of explants:leaf,stems,petiole,and root.The potency to form callus varies between different explants and leaf explants exhibits the highest capacity(100%).On the other hand,root-derived callus(R-callus)produces the highest level of total amount of PF and AF,31.8 mg·g-1dry mass,which is higher than the corresponding level in the root of field cultivated PL.Furthermore,the time needed is only 40 days,remarkably shorter than the cultivation time of PL,about 4–5 years.Higher accumulation levels of PF and AF with shorter production time indicate that callus culture of PL is a promising powerful tool for production of PF and AF in the future.

Pharmacological effects of Paeoniflorin and Albiflorin on IL-3, GM-CSF, IL-6 and TNF-α in the rats of syndrome of stagnation of liver qi and blood deficiency

Objective： To observe the effect of paeoniflorin （PF）, albiflorin （AF） on the hemogram, visceral index and hematopoiesis cytokine in the rats of syndrome of stagnation of liver qi and blood deficiency, and to discuss the material base and mechanism of effect of nourishing blood and smoothing the liver of Baishao （Radix Paeoniae Alba）. Methods： Male SD rats were randomly divided into groups according to the sucrose preference test and body weight （n = 12）. Except the normal control, the other groups were treated with the chronic stress stimulation combined with radiation respectively to establish the model of syndrome of stagnation of liver qi and blood deficiency. The body weight, visceral index and the quantity of Leucocyte, Red Blood Cells, Hemoglobin in peripheral hemogram were monitored, then plasma and serum were separated. Radioimmunoassay was used to analyze the levels of Lnterleukin-3, Granulocyte-macrophage Colony-stimulating Factor, Lnterleukin-6 and Tumor Necrosis Factor-α in plasma. Results： Compared with that of model group, 30 mg·kg^-1 PF and 30 mg·kg^-1 AF of the weight, spleen index, quantity of Leucocyte were increased significantly （P 〈 0.05, P 〈 0.01）. The results of Radioimmunoassay showed that the levels of Interleukin-3 increased （P 〈 0.05, P 〈 0.05） and the levels of Tumor Necrosis Factor-α decreased in both 30 mg·kg^-1 PF and 30 mg·kg^-1 AF groups （P 〈 0.05, P 〈 0.05）. Conclusion： The effect of PF and AF on the regulation of bone marrow hematopoietic system and immune system play a role in the blood of rats with syndrome of stagnation of liver qi and blood deficiency, which suggests that both of them are the main active ingredients of nourishing blood and smoothing the liver of Baishao.

Albiflorin attenuates inflammatory injury by regulating the TLR4 signaling pathway and its negative regulating factor Tollip in experimental models of ulcerative colitis

Albiflorin （AF） is the main active component extracted from Paeoniae Radix Alba. This study investigated the efficacy of AF in attenuating inflammatory injury by regulating the TLR4 signaling pathway and its negative regulating factor Tollip in an experimental ulcerative colitis （UC） model. We administrated trinitrobenzene sulfonic acid for 21 d to induce UC in rats. The efficacy of AF in attenuating UC was assessed using various biochemical markers, such as tumor necrosis factor-α （TNF-α）, interleukin- 1 （IL- 1）, interleukin- 10 （IL- 10）, 5-hydroxytryptamine （5-HT）, and tissue myeloperoxidase （MPO）, along with histopathological studies on toll-like receptor-4 （TLR-4） signaling pathway and its negative regulating factor Tollip. The results showed that AF can significantly downregulate the levels of TNF-α, IL-1, IL-10, and 5-HT. AF decreased the activation of TLR4, MyD88, and NF-κB p65 protein expression by increasing Tollip expression. AF can relieve symptoms of UC by suppressing the activation of the TLR4 signaling pathway and upregulating its negative regulating factor Tollip. Therefore, AF may be a potential natural product for treating UC.

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.

Insights into the neutral loss of 30 Da of Paeonia monoterpene glycosides in electrospray ionization tandem mass spectrometry and the rapid screening of monoterpene glycosides by LC/MS/MS

Monoterpene glycosides are the major bioactive compounds of Paeonia lactiflora Pall（P. lactiflora）. Characteristic neutral loss of 30 Da has been extensively reported for monoterpene glycosides in tandem mass spectrometry. However, little is known about mechanism of this fragmentation. The neutral loss of 30 Da was studied for eleven monoterpene glycosides（1–11） from P. lactiflora by ion trap mass spectrometry in this report. Compounds 1–5 with a hemiacetal structure could readily lose 30 Da at low collision energy of 30% in MS/MS by ion trap mass spectrometry. For compounds 6–11, neutral loss of 30 Da could also be observed at low abundance, but the collision energy had to be increased to 60%. In both cases, high-accuracy mass spectrometry assigned the 30 Da as CH_2O. After careful analysis of the structures and mass spectra, we believe that the neutral loss of 30 Da in compounds 1–5 was due to cleavage of the hemiacetal structure, whereas it was ascribed to the cleavage of 5＇-hydroxymethyl group of the glucosyl residue in other monoterpene glycosides. Furthermore, the characteristic neutral loss of 30 Da at low collision energy was used to screen hemiacetals from crude extracts of P. lactiflora and related plant species. Significant differences among Paeonia species were observed by 30 Da neutral loss analysis.

Sedative and hypnotic effect of freeze-dried paeoniflorin and Sini San freeze-dried powder in pentobarbital sodium-induced mice

OBJECTIVE: To investigate the sedative and hypnotic activity of paeoniflorin and freeze-dried Sini San powder on mice and provide a reliable method for determining the pharmacodynamic material basis of Sini San.METHODS: Male adult mice weighing 20-22 g were used in this study. Three experiments were carried out. Synergism with pentobarbital was used as an index for hypnotic effect. Loss of the righting reflex was used to determine the start of sleep.Sleep latency and sleeping time were recorded in each experiment.RESULTS: The coefficient of variation of the suprathreshold dose(55 mg/kg) was significantly lower than that of the threshold dose. The sleep latency of mice was significantly decreased, and the sleeping time of mice was significantly prolonged. The effects of paeoniflorin and Sini San on prolonging the sleeping time of mice induced by pentobarbital sodium were significantly stronger than those in the control group.CONCLUSION: Paeoniflorin produces significant sedative and hypnotic effects, and there is an obvious dose-effect relationship.