Ischemic stroke is a significant public health problem worldwide. Growing evidence has shown gut microbiota served a vital function in ischemic stroke. Rhubarb, always used after processing, is a promising therapy for ischemic stroke. However, the possible mechanism of rhubarb with different cooking methods has remained unclear. Herein, the constitutes of steaming rhubarb (SP), raw rhubarb (RP), and nine steaming nine sun-drying rhubarb (NSP) were identified via LC-QTOF-MS. The middle cerebral artery occlusion (MCAO) mice model was constructed. Infarction area, neurological score, Nissl staining, IBA-1 immunofluorescence, and ELISA were performed to confirm the neuroprotective effect of SP, RP, and NSP. The gut microbiota in fetal was studied via 16sRNA sequencing, and the level of short-chain fatty acids (SCFAs) in brain and gut were measured via GC-MS. The function of microbiota signature was identified through PICRUSt2; the possible mechanism was studied. SP, RP, and NSP alleviated the neurological dysfunction, decreased the inflammation, suppressed dysbiosis of gut microbiota, restored SCFA-producing bacteria, and enhanced the SCFA level in MCAO mice. Moreover, the NSP and SP enriched the proportion of anti-inflammation and beneficial bacteria, deleted the proportion of pro-inflammation. It observed energy metabolism was involved in the possible mechanism of rhubarb; activities of COXI and Na+-K+-ATPase were increased in the brain of NSP and SP treatment mice. Furthermore, the expression of GLUT4 and PFK1 (the energy metabolism-related genes) was elevated in the brain after RP, NSP and SP administration. In this study, it provided proof for the treatment of ischemic stroke with rhubarb. Rhubarb restored the gut microbiota and regulated the expression of GLUT4 and PFK1 to alleviate ischemic stroke.

To explore the effect of the granules made by new-medicinal parts of Crocus sativus(NMPCS) on hyperuricemia(HUA) in rats, the rat model of HUA was established by intramuscular injection of 3% potassium oxonate and intraperitoneal injection of 4% pyrazinamide. The content of serum uric acid was monitored every week for 3 consecutive weeks. After the experiment, the levels of serum uric acid, urine uric acid, serum creatinine, blood urea nitrogen(BUN), and xanthine oxidase(XOD) were determined. The protein and gene expressions of XOD were determined by Western blot method and fluorescence quantitative polymerase chain reaction(qPCR), and the morphological changes in the liver tissue were performed by hematoxylin-eosin(HE) staining. The results showed that as compared with the model group, the levels of serum uric acid in the positive drug group and the low, medium, and high-dose NMPCS groups were lower(P<0.05), the levels of urine uric acid in the high-dose NMPCS group were decreased(P<0.01), and there was no statistical difference in the medium and low-dose NMPCS groups. The levels of BUN in the high and low-dose NMPCS groups were decreased(P<0.05), and the levels of serum creatinine did not change in the administration groups. The positive drug group and the low, medium, and high-dose NMPCS groups significantly reduced the liver damage, with only a few hepatocytes vacuolization and a small number of red blood cells in the central venous area. The nephridial tissue structure was slightly abnormal, with a small number of red blood cell infiltration, and no obvious inflammatory cell infiltration was found in the glomerulus in these groups. No degeneration was found in renal tubular epithelial cells, with mild glomerular and tubular lesions and a small amount of sodium urate deposition and crystallization in the positive drug group and the low, medium, and high-dose NMPCS groups. The relative protein expressions of XOD in the positive drug group and the high dose NMPCS group were decreased(P<0.05), and the relative mRNA expressions of XOD in the positive drug group and the high and low-dose NMPCS groups were decreased as well(P<0.05). The above results show that NMPCS reduces uric acid in rats with HUA by regulating XOD, which provides a certain experimental basis for the development of NMPCS as a new medicine for the treatment of HUA.

Only the dried stigma of the saffron, a flower deemed as the most valuable spice globally, is utilized for industrial production. Hence, there exists a growing interest in utilizing saffron floral bio-residues. The anti-hyperuricemic activity of a flavonoid extract from saffron floral bio-residues was assessed in potassium oxonate-induced hyperuricemia mice. In addition, an ultra-high performance liquid chromatography-triple quadrupole mass spectrometry method was established and validated to determine the pharmacokinetics of five main flavonoids and three phase-II metabolites in rat plasma after oral administration of the flavonoid extract for the first time. Compared with pharmacokinetic parameters of kaempferol-3-O-sophoroside, the most abundant flavonoid in the extract, and its aglycone kaempferol, we observed that coexisting compounds significantly reduced the absorption, accelerated the excretion of kaempferol-3-O-sophoroside, while significantly increasing the absorption and prolonging the residence time of kaempferol in the flavonoid extract. These results suggest the promising potential of the flavonoid extract from saffron floral bio-residues as an anti-hyperuricemic agent. Kaempferol was absorbed in plasma at high concentrations owing to the biotransformation of kaempferol glycosides in vivo.

Background: In our previous study, we found that the combination of a traditional Chinese medicine (TCM) and swimming could prevent atherosclerosis through a synergistic interaction. However, whether the combined application of active components from the fruit of Crataegus pinnatifida Bge. Var. major N.E. Br. and the root of Salvia miltiorrhiza Bge. (CPSM) and swimming has been effective in the prevention and treatment of focal cerebral infraction remained unclear. This work aimed to conduct detailed investigation on the joint effects of CPSM extract with swimming on focal cerebral infraction in rats and its underlying mechanisms. Method: A photochemical method of the combination of Rose Bengal (RB) injection and cold-light source irradiation was performed to establish the rat focal cerebral thrombosis model. The pathological changes of the brain were observed by a DCP-7030 laser multifunction machine, and the protein levels of von Willebrand factor (vWF), vascular cell adhesion molecule-1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1) were detected by Western blotting. Blood samples were collected to assay tissue plasminogen activator (t-PA), plasminogen activator inhibitor type-1 (PAI-1), endothelin-1 (ET-1), 6-keto-prostaglandin F1α (6-keto-PGF1α), and thromboxane B2 (TXB2). Finally, the gene expression of t-PA, PAI-1, and ICAM-1 in human umbilical vein endothelial cells (HUVECs) stimulated by tumor necrosis factor-α (TNF-α) was assayed via real-time (RT) quantitative PCR (qPCR). Results: The joint effects of CPSM extract and swimming demonstrated significant interactions, which including increased blood perfusion, increased serum t-PA and 6-keto-PGF1α, decreased serum PAI-1 and TXB2, decreased protein levels of vWF, VCAM-1 and ICAM-1, and decreased ICAM-1 gene expression. Conclusion: This research demonstrated that the combined therapy of CP and SM extracts with swimming could prevent focal cerebral infraction through interactions on the regulation of vascular endothelial functions and inflammatory factors. It stresses the promising effects of the drugs and shear stress of blood flow in prevention and treatment of thrombosis. The mechanism may be related to regulating the protein expression of vWF, VCAM-1, and ICAM-1, and downregulating the gene expression of ICAM-1.

目的:研究炒莱菔子醇提物中芥子酸在正常大鼠体内的药代动力学特征以及在哮喘大鼠病理状态下的药代动力学-药效动力学(PK-PD)相关性.方法:正常大鼠分别单次灌胃低、中、高剂量(4.5,9,18 g·kg-1)的炒莱菔子醇提物,利用UHPLC-MS/MS测定不同时间点血浆中芥子酸含量,采用DAS 3.2.8软件进行数据分析以获取药代动力学参数;采用腹腔注射卵清蛋白佐以氢氧化铝建立大鼠哮喘模型,分别灌胃低、高剂量(4.5,9 g·kg-1)炒莱菔子醇提物,3周后眼底静脉丛取血,分离血浆和血清,UHPLC-MS/MS测定不同时间点血浆中芥子酸含量,利用酶联免疫吸附测定法(ELISA)测定血清中免疫球蛋白E(IgE),白细胞介素-5(IL-5),肿瘤坏死因子-α(TNF-α)活性,采用DAS 3.2.8软件进行PK-PD模型拟合以及数据分析.结果:正常大鼠灌胃给予低、中、高剂量炒莱菔子醇提物后,血浆中芥子酸药峰浓度(Cmax)分别为(29.35±10.32),(62.70±27.47),(137.33±40.95) μg·L-1;药时曲线下面积(AUC0-t)分别为(92.83±27.16),(240.74±75.09),(633.95±195.88) μg·L-1·h;达峰时间(Tmax)分别为(2.58±0.80),(3.00±0),(5.50±1.23)h;与炒莱菔子醇提物低剂量组比,炒莱菔子醇提物中、高剂量组的AUC0-t,平均滞留时间(MRT0-t)均显著增加(P ＜0.05,P＜0.01);芥子酸体内AUC0-t在4.5～18 g·kg-1线性依赖性良好.哮喘大鼠分别灌胃低、高剂量炒莱菔子醇提物后0.083,0.167 h,与模型组比较,血清中IL-5,IgE,TNF-α含量均有不同程度地降低(P＜0.05,P＜0.01);血浆中芥子酸Cmax分别为(58.43±27.94),(61.16±18.79) μg·L-1;AUC0-t分别为(188.75±37.07),(247.90±36.89) μg· L-1·h;AUC0-t,表观分布容积(Vz/F)和清除率(CLz/F)均随剂量增加而增加,最佳药动学模型拟合为血管外给药的一房室模型,PK-PD模型可能适用于间接连接模型.结论:哮喘大鼠体内芥子酸血药浓度与血清中IL-5,IgE,TNF-α含量存在一定的相关性,给药剂量和机能状态(病理或生理状态)均会影响炒莱菔子中芥子酸在大鼠体内的药代动力学行为,并与其平喘药效有一定的关联性.

In this research, high-throughput sequencing was used to investigate the mechanism of Naoxintong Capsules(NXTC) in prevention of post-ischemic inflammation. First, microglia BV-2 inflammatory model was induced by 1.0 μg·mL~(-1) LPS to investigate the effect of intestinal absorption solution of NXTC(NXTCIA) at different concentrations(62.5, 31.25, 15.63, 7.81 μg·mL~(-1)) on LPS-induced BV-2 inflammatory factors in microglia. Then, an RNA-Seq high-throughput sequencing method was performed to identify the differentially expressed mRNAs in microglia BV-2 after pre-treatment with NXTC. GO and KEGG enrichment analysis was used to screen the potential biological processes and related signaling pathways of NXTC in inhibiting inflammation. The results showed that four NXTCIA concentrations could significantly inhibit the release of LPS-induced inflammatory mediators in BV-2 in a dose-dependent manner. Furthermore, high-throughput sequencing results showed that 392 mRNA transcripts were reversed following pre-treatment with NXTC. GO enrichment analysis showed that the transcripts reversed by NXTC were mainly involved in Toll-like receptor signaling pathway, chemokine signaling pathway, and TNF signaling pathway. Taken together, our findings showed that NXTC treatment could provide protective effects against inflammatory response and the mechanism might be related to the regulation of Toll-like receptor signaling pathway, chemokine signaling pathway, and TNF signaling pathway.

Xueshuantong capsule (XST) is a patented traditional Chinese medicine used for the prevention and treatment of thrombosis. The molecular mechanism of anti-thrombotic effect of XST was investigated through the cross-talk among the platelets/leukocytes, endothelial cells (ECs), and flow shear stress. The Bioflux 1000 system was used to generate two levels of shear stress conditions: 0.1 and 0.9 Pa. Bioflux Metamorph microscopic imaging system was used to analyze the adhesion cell numbers. Protein expressions were detected by western blotting and flow cytometry. The flow-cytometry results showed that under 0.1 Pa flow, XST decreased ADP induced platelets CD62p surface expression in a concentration-dependent manner. Under 0.9 Pa flow, XST at a concentration of 0.15 g⋅L-1 reduced the platelets activation by 29.5%, and aspirin (ASA) showed no inhibitory effects. XST showed similar efficiency on monocytes adhesion both under 0.1 and 0.9 Pa flow conditions, and the inhibition rate was 30.2 and 28.3%, respectively. Under 0.9 Pa flow, the anti-adhesive effects of XST might be associated with the suppression of VE-cadherin and Cx43 in HUVECs. Blood flow not only acts as a drug transporter, but also exerts its effects to influence the pharmacodynamics of XST. Effects of XST on inhibiting platelets activation and suppressing platelets/leukocytes adhesion to injured ECs are not only concentration-dependent, but also shear stress-dependent. The mechanic forces combined with traditional Chinese medicine may be used as a precise treatment for cardiovascular diseases.

Objective: To investigate the antiplatelet aggregation effect of water-soluble tomato concentrate (WSTC) and explore the underlying molecular mechanisms. Materials and Methods: Platelet aggregometry was used to quantify rat platelet aggregation with the maximum aggregation rate in vitro and ex vivo. Then, the fibrinogen (FIB) binding assay was employed to detect the effect of WSTC on the activation of platelet integrin αIIβ3 (GP IIb/IIIa). Furthermore, Western blot was performed to assess the platelet protein levels of phosphoinositide 3-kinase 110 β (PI3K110 β), protein disulfide isomerase (PDI), platelet endothelial cell adhesion molecule 1 (PECAM-1), and β1-Tubulin. Results: WSTC inhibited adenosine diphosphate (ADP) and collagen-induced platelet aggregation in a concentration-dependent manner in vitro, at IC50values of 3.05 g/L and 8.03 g/L, respectively. Significantly reduced ex vivo ADP induced platelet aggregation was observed after oral consumption of WSTC for 4 weeks in rats; average inhibition rates were 24.42%, 21.48%, and 20.87% for 25 mg/Kg, 75 mg/Kg, and 150 mg/Kg WSTC, respectively. It appeared that WSTC had no influence on coagulation function in rats. Incubation with WSTC decreased FIB binding to GP IIb/IIIa by 17.47% and 32.29% at the concentrations of 0.6 and 6 g/L, respectively. WSTC at 0.6 and 6 g/L markedly downregulated PI3K110 β, PDI, and PECAM-1 in platelets, and upregulated β1-Tubulin, in a concentration-dependent manner. Conclusion: WSTC inhibits platelet activation through modulation of platelet skeletal stability and suppresses GP IIb/IIIa receptor-mediated platelet aggregation, likely via the PI3K signaling pathway and PDI inhibition.

Objective: To establish a method coupled with everted gut sacs and isolated vascular, and study the effects of Yindan Xinnaotong Capsule on intestinal absorption of vasoactivity. Methods: The everted intestinal sac method was performed to prepare the intestinal absorption solution. The isolated rat aortic ring model was used to observe the effects of intestinal absorption on the vasoactivity in vitro. Additionally, the main compounds (ginsenoside Rg1, ginsenoside Rb1, ginsenoside Re, ginsenoside Rd, notoginsenoside R1, ginkgolide A, ginkgolide B, ginkgolide C, bilobalide, rutin, and tanshinone IIA) in intestinal absorption solution were determined by UPLC-QQQ. Results: The intestinal absorption liquid of Yindan Xinnaotong Capsule in the concentration range of 100-1 600L had a concentration-dependent vasodilation. Eleven constituents could be absorbed by gut sacs. The cumulative amount of intestinal absorption of all components was the highest in high concentration and the lowest in low concentration. Conclusion: The method is simple and controllable. It can reflect the true biological activity of samples, which provide a reference for the quality evaluation of Yindan Xinnaotong Capsule.

ETHNOPHARMACOLOGICAL RELEVANCE:The danshen (the root of Salvia miltiorrhiza Bge.)-shanzha (the fruit of Crataegus pinnatifida Bge. var. major N.E.Br.) (DS) herb combination is a commonly used traditional Chinese medicine with cardiovascular disease (CVD) treatment potential. MATERIALS AND METHODS:In this study, we investigated the anti-atherosclerotic effects and mechanisms of DS by the integration of network pharmacology and polypharmacology. Eight main components were selected for target fishing by PharmMapper. RESULTS:The network pharmacological study indicated that DS may target 41 proteins and 16 pathways associated with inflammation, lipid metabolism and endothelial protection, which indicates that DS probably adjusts these processes as part of its anti-atherosclerotic activities. Furthermore, this hypothesis was verified by polypharmacology using an atherosclerotic model. Histopathological examination showed that DS inhibited pathological changes in the arteries of atherosclerotic rats and reduced the intima-media thickness (IMT). DS significantly reduced the levels of total cholesterol (TC), triglyceride (TG), and low-density lipoprotein-cholesterol (LDL-C) and increased the high-density lipoprotein-cholesterol (HDL-C) level in the blood. DS also decreased the concentrations of interleukin (IL)-1β and IL-18, indicating anti-inflammation activity. In addition, DS increased the serum levels of nitric oxide (NO) and 6-keto-prostaglandin F1α (6-keto-PGF1α) and decreased the serum levels of endothelin (ET) and thromboxane B2 (TXB2), indicating an endothelial protective effect. CONCLUSIONS:In conclusion, DS has an anti-atherosclerotic ability to lower lipid concentrations and to protect endothelial function, and it also has anti-inflammatory activity.

Background: It has been recognized that exercise training can attenuate the progression of atherosclerosis (AS). The combined application of components from the fruit of Crataegus pinnatifida Bge. Var. major N.E. Br. (CP) and the root of Salvia miltiorrhiza Bge. (SM) has been effective in the prevention and treatment of atherosclerosis. The present work aims to investigate the joint effects of extracts from the fruit of CP and the root of SM with swimming on atherosclerosis in rats. Method: To establish a rat atherosclerosis model, a combined method of partial ligation of the left common carotid artery leading to low shear stress and a high-fat diet was employed. Blood samples were collected to detect the blood lipid profile, which included total cholesterol (T-CHO), low-density lipoprotein cholesterol (LDL-C), triglycerides (TG) and high-density lipoprotein cholesterol (HDL-C); endothelial cytokines such as 6-keto-prostaglandin F1α (PGF1α), endothelin (ET), thromboxane B2 (TXB2), plasminogen activator inhibitor-1 (PAI-1), and von Willebrand factor (vWF); and inflammatory cytokines such as interleukin-1β (IL-1β), interleukin-6 (IL-6) and interleukin-10 (IL-10). Finally, the common carotid arteries of the rats were removed to observe pathological changes via oil red O staining, and the gene expression of t-PA, PAI-1, and vWF was assayed via real-time (RT) quantitative polymerase chain reaction (qPCR). Results: The joint effects of CPSM extract and swimming indicated significant interactions, including (1) decreased serum T-CHO, TG, and LDL-C; (2) decreased IL-6 and increased IL-10; (3) decreased TXB2, PAI-1 and vWF; three-dimensional analysis showed that gene expression of PAI-1 was inhibited, vWF gene expression was downregulated, and COX-1 gene expression was increased; and (4) decreased lipoprotein retention in the carotid artery wall. Conclusion: This research demonstrates that the combined therapy of CP and SM extracts with swimming can improve blood lipid levels and endothelial functions and attenuate the early signs of atherosclerosis in a rat model of atherosclerosis. The regulation of the gene expression of PAI, vWF and COX-1 may be the underlying cause of the effect. Methodologically speaking, three-dimensional surface plots of the joint effects of CPSM extract and swimming on parameters with quadratic fitting yielded a more accurate equation for describing the dose-response relationship in biomechanopharmacology. Such plots are likely worth using in pharmacology to quantify the effects induced by two medicinal factors.

目的:研究三黄泻心汤活血化瘀优势方对氧化低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞(HUVECs)损伤的影响并探讨其抗动脉粥样硬化的作用机制.方法:采用体外培养的HUVECs,分为空白组,ox-LDL损伤组(模型组),三黄泻心汤活血化瘀优势方15.63,31.25,62.50 mg· L-1组;用200 mg·L-1 ox-LDL孵育24 h,造成HUVECs损伤模型;细胞存活率采用噻唑蓝(MTT)比色法测定,放射免疫法检测细胞上清液中内皮素(ET-1),6-酮-前列腺素F1α(6-keto-PGF1α),血栓烷素B2(TXB2)含量,半自动生化分析仪检测抗氧化应激指标超氧化物歧化酶(SOD),丙二醛(MDA),一氧化氮(NO)水平.采用试剂盒荧光定量法检测活性氧(ROS)含量,检测细胞间黏附分子-1(ICAM-1),血管细胞黏附分子-1(VCAM-1)及单核细胞趋化因子-1(MCP-1)水平酶联免疫吸附测定.细胞凋亡率及线粒体膜电位变化采用流式细胞术,采用蛋白免疫印迹法(Western blot)检测凋亡相关蛋白B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2),Bcl-2相关X蛋白(Bcl-2-associated X protein,Bax),半胱氨酸蛋白酶-3(cysteine aspartate-specific protease-3,Caspase-3),Caspase-9表达.结果:与空白组比较,模型组HUVECs存活率降低;MDA,MCP-1,ICAM-1,VCAM-1的水平升高,SOD和NO含量降低,ROS的分泌增加,细胞凋亡率升高,Bax,Caspase-3和Caspase-9的表达量增加,Bcl-2的表达量减少(P＜0.05,P＜0.01).与模型组比较,不同质量浓度的三黄泻心汤活血化瘀优势方使HUVECs存活率明显上升,显著降低MDA,MCP-1,ICAM-1,VCAM-1的水平;提高SOD和NO含量,且减少ROS的分泌,降低细胞凋亡率,降低Bax,Caspase-3和Caspase-9的表达量,增加Bcl-2的表达量(P＜0.05,P＜0.01).结论:在ox-LDL诱导损伤的HUVECs模型上,三黄泻心汤活血化瘀优势方具有抑制细胞凋亡,减轻炎性反应和抗氧化的作用,可能与抗动脉粥样硬化有密切关系.

Yindanxinnaotong capsule (YDXNT), a traditional Chinese formula, has been used to treat cardio-cerebrovascular diseases for several decades. Previous research has focused on evaluating the pharmacological properties and main compounds of YDXNT in vitro and in vivo. However, the multiple bioactive compounds in vivo remain poorly understood. In the present research, an integrative strategy using UPLC-Q-TOF-MS combined with UPLC-QqQ-MS was employed to detect the absorbed constituents and investigate the pharmacokinetics of main compounds in the plasma after oral administration of YDXNT. UPLC-Q-TOF-MS was developed to detect the absorbed constituents and their metabolites in the plasma after oral administration in rats. A total of 52 constituents, including 44 prototype compounds and 8 metabolites, were identified or tentatively characterized. Then, nine main compounds (quercetin, isorhamnetin, kaempferol, ginkgolide A, ginkgolide B, ginkgolide C, bilobalide, tanshinone IIA, and salvianolic acid B) were chosen to further investigate the pharmacokinetic behavior of YDXNT using UPLC-QqQ-MS. The concentration of nine main constituents were in the range of 27.85-76.54 ng mL(-1). This research provides a systematic approach for rapid qualitative analysis of absorbed constituents and for evaluating the pharmacokinetics of the main ingredients of YDXNT following its oral administration. More importantly, this work provides key information on the identification of bioactive compounds and the clarification of their action mechanisms, as well as on the pharmacological actions of YDXNT.

目的 研究流动剪应力与活血化瘀药物丹参山楂提取物联合应用抗血栓的作用机制.方法 采用生物力药理学的研究方法,利用BioFlux1000控剪应力微流培养系统,分别施加0.1、0.9 Pa剪应力并联合不同剂量丹参山楂提取物对血管内皮细胞(HUVECs)进行预处理后,用ox-LDL诱导内皮细胞损伤,采用酶联免疫吸附法(ELISA)检测细胞上清液中PAI-1、t-PA、ICAM-1的含量;采用RT-PCR法分别测定PAI-1、t-PA、ICAM-1基因表达.结果 丹参山楂提取物能下调内皮细胞中PAI-1 mRNA的相对表达,减少内皮细胞中PAI-1的分泌;与低剪应力相比,正常剪应力联合大剂量丹参山楂提取物能够上调内皮细胞中t-PA mRNA的表达,增加内皮细胞分泌t-PA;在低剪应力条件下,丹参山楂提取物可明显抑制ICAM-1的表达.结论 正常水平剪应力联合丹参山楂提取物能够通过调节纤溶系统的稳定性来抑制血栓形成;丹参山楂提取物能够通过抑制内皮细胞中PAI-1的表达产生抗血栓效应;在病理剪应力条件下,丹参山楂提取物对抑制ICAM-1表达的药效更加显著.

目的 研究家兔离体动脉血管段在不同剪应力和不同平均压力组合作用下分泌PAI-1、t-PA、vWF水平及基因表达水平的规律.方法 手术分离实验家兔腹主动脉、颈总动脉,采用离体血管灌流装置,对家兔动脉血管实施不同剪应力(范围2～22 dyn·cm-2)和不同压力(范围9.0～48.4 mmHg);放免法测定灌流剪切液中vWF、PAI-1、t-PA的含量;RT-PCR法测定应力加载动脉血管vWF,PAI-1和t-PA基因的表达水平.结果 低水平剪应力联合低压力,高水平剪应力联合高压力,中等水平剪应力联合中等压力3个组合作用于内皮细胞,可抑制PAI-1的表达;中等水平剪应力和中等压力联合作用,可促进t-PA的表达;高水平的血流剪应力与高压力联合作用,可抑制vWF的表达.结论 离体家兔动脉血管灌流时,血流剪应力与压力联合作用可影响血管内皮细胞PAI-1,t-PA,vWF的分泌,其作用机制则与应力调节相应基因的表达有关.

Guanxinjing capsules (GXJCs) are used in traditional Chinese medicine as a common therapy for coronary heart disease (CHD) complicated with depression. In this study, we aimed to identify the main active constituents in GXJCs and to investigate the mechanisms of GXJC action on CHD complicated with depression. The chemical constituent profile of the GXJC was identified by UHPLC-LTQ-Orbitrap assay, and oral bioavailability was evaluated to screen the GXJC drug-like chemical constituents. A total of 16 GXJC drug-like chemical constituents were identified. Then, putative targets of the GXJC drug-like chemical constituents were predicted using MedChem Studio, with 870 genes found to be the putative targets of these molecules. After that, a GXJC putative target-known CHD/depression therapeutic target network was constructed, and four topological features, including degree, betweenness, closeness and K-coreness, were calculated. According to the topological feature values of the GXJC putative targets, 14 main active constituents were identified because their corresponding putative targets had topological importance in the GXJC putative target-known CHD/depression therapeutic target network, which were defined as the candidate targets of GXJC against CHD complicated with depression. Functionally, these candidate targets were significantly involved in several CHD/depression-related pathways, including repairing pathological vascular changes, reducing platelet aggregation and inflammation, and affecting patient depression. This study identified a list of main active constituents of GXJC acting on CHD complicated with depression using an integrative pharmacology-based approach that combined active chemical constituent identification, drug target prediction and network analysis. This method may offer an efficient way to understand the pharmacological mechanisms of traditional Chinese medicine prescriptions.

To observe the protective effect of scutellarin ethyl ester on focal cerebral ischemia injury induced by middle cerebral artery occlusion in rats(MCAOR), and explore its mechanism. Totally 84 male SD rats were randomly divided into seven groups： sham-operated group, model group,positive drug group(niomdipine,12 mg•kg⁻¹), Brevisapin tablets group(48 mg•kg⁻¹), and high, middle and low-dose scutellarin ethyl ester groups(100, 50, 25 mg•kg⁻¹). The MCAOR model was prepared by using thread embolism method to observe the neurological function of rats, the area of cerebral infarction was measured with TTC, and the levels of MDA, SOD and NO in serum were detected with semiautomatic biochemistry analyzer.Ox-LDL and TNF-α cell injury models was established by treating HUVECs with 200 mg•L⁻¹ ox-LDL and 100 μg•L⁻¹ TNF-α,and the levels of MDA, SOD, NO, ET, 6-keto-PGF1α,TXB2, IL-1, IL-6, IL-8, ICAM-1 and PECAM-1 in the cell supernatant were determined. The results showed that scutellarin ethyl ester could effectively improve the neurological function of MCAOR rats, and significantly reduce the area of cerebral infarction. Compared with the model group, activities of SOD and NO in serum increased, while content of MDA decreased. In the cell supernatant, activities of SOD, 6-keto-PGF1α and NO increased, content of IL-1, IL-6, IL-8, ICAM-1, PECAM-1, TXB2, ET and MDA decreased, which indicated that scutellarin ethyl ester has a certain protective effect on focal cerebral ischemia injury induced by middle cerebral artery occlusion in rats, and its mechanism may be related to antioxidative stress, improvement of endothelial function and reduction in inflammatory reaction.

To estabish ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for simultaneous determination of quercetin(QCT), isorhamnetin(ISR), kaempferol(KMF), ginkgolide A(GA), ginkgolide B(GB), ginkgolide C(GC) and bilobalide(BB) in rat plasma and investigate the pharmacokinetic process of seven compounds after oral administration of Yindan Xinnaotong Ruanjiaonang, The results indicated that all calibrations curves showed good linearity (r≥0.997 1). RSD of intra-day and inter-day precisions were all within 11%. The matrix effects and extraction recovery were in the range of 93.28%-103.6% and 72.43%-95.77% respectively. The peak concentration (Cmax) of QCT, ISR, KMF, GA, GB, GC and BB were (45.02±11.28), (49.90±13.82), (27.85±8.38), (76.31±18.19), (76.54±15.43), (35.35±10.28), (48.70±12.34) μg•L⁻¹, respectively. The peak time (tmax) of seven constituents were (0.33±0.11), (0.50±0.23), (0.33±0.14), (0.75±0.29), (1.0±0.35), (1.5±0.23), (0.75±0.50) h, respectively. UPLC-MS/MS method established in this research was proved to be so rapid and sensitive that it can be applied to the pharmacokinetic study of seven bioactive constituents in Yindan Xinnaotong Ruanjiaonang.

Objective To study the effects of microemulsion/ethosomes on transdermal absorption properties and efficacy of Huoxue Zhitong Cataplasm. Methods The improved Franz diffusion cells were used for the in-vitro permeation experiment with rat skins as the barriers, which was used to evaluate the transdermal absorption properties. In the erxeriment, the contents of paeonol, eugenol and methyl salicylate were used as markers, and detected by ultra performance liquid chromatography to evaluate the transdermal absorption effects. The anti-inflammatory and analgesia activity were evaluated through the writhing plate experiments. Results The cumulative release rate of paeonol in Huoxue Zhitong Cataplasm, Microemulsion Huoxue Zhitong Cataplasm and Ethosomes Huoxue Zhitong Cataplasm were, in order, 65.30%, 61.30%and 60.20%in 24 h;eugenol were, in order, 51.08%, 54.71% and 55.66% in 24 h; methyl salicylate were, in order, 49.20%, 65.17% and 72.15% in 24 h. Furthermore, Microemulsion Huoxue Zhitong Cataplasm high-dose group and Ethosomes Huoxue Zhitong Cataplasm medium-dose group had good effects on reducing the inflammatory exudate of peritoneal capillary and capillary permeability (P<0.05) in animal models. Conclusion Huoxue Zhitong Cataplasm based on microemulsion/ethosomesnano-technology has good transdermal absorption properties and efficacy.