The UHPLC-LTQ-Orbitrap high resolution mass spectrometer was used to explore the chemical compositions in safflower. The rapid separation of the compositions was conducted by the UHPLC, following by high resolution full scan and MS2 scan, under the positive and negative ion mode. The chemical formula of compositions were deduced by full scan data in less than 5, then the potential structures were confirmed by the MS2 data. Forty-nine compounds were detected, of which 26 was identified, and 5 compounds was validated by the standard substances.
Carthamus tinctorius ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Tandem Mass Spectrometry ; methods

Gushukang is a compound Chinese herbal preparation. Pharmacological studies indicated that Gushukang can increase bone density, inhibit bone resorption, promote bone formation, and restore bone microstructure, as well as improve bone biomechanical parameters and promote healing of bone fracture. Clinical observations demonstrated that it has favorable efficacy in preventing and treating osteoporosis.
Bone Density ; drug effects ; Bone Resorption ; prevention & control ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Osteogenesis ; drug effects ; Osteoporosis ; drug therapy ; physiopathology ; Phytotherapy

To explore the prevention effect of the joint combination of Yindanxinnaotong soft capsule (YDXNT) and exercise (swimming) on atherosclerotic rats. The method of 3 x 3 factorial design, including two factors (YDXNT and swimming) and three levels (0, 1, 2 g x kg(-1) YDXNT; 0, 0.5, 1 h swimming), was mainly adopted. The atherosclerotic rat model was established by ligating their left common carotid arteries and feeding high-fat diet. After 8 weeks, blood samples were collected from their thoracic aorta to determine blood viscosity, plasma viscosity, fibrinogen (FIB), nitric oxide (NO), 6-keto-PGF(1alpha) endothelin (ET) and thromboxane B2 (TXB2). The tissues of left common carotid arteries of the rats were collected to detect the positive expression of SM22alpha and determine the semi-quantitation through the immunohistochemical staining. The result showed that the combination of YDXNT and swimming can significantly decrease the plasma viscosity (F = 3.241, P = 0.017), the high and low shear blood viscosity (F = 6.444, P = 0.001; F = 3.002, P = 0.024) and FIB (F = 4.046, P = 0.005). The increased NO and 6-keto-PGF(1alpha) and the decreased ET and TXB2 indicated a significant interaction (P < 0.05). The swimming showed an obvious main effect in the expression of up-regulated protein SM22alpha (F = 8.088, P = 0.001). The study suggested that the combined administration of YDXNT and swimming could improve the hemorheological parameters of atherosclerotic rats, protect the vascular endothelium, inhibit the vascular remodeling in atherosclerosis and positively prevent the atherosclerosis.
Animals ; Atherosclerosis ; drug therapy ; genetics ; metabolism ; prevention & control ; Blood Viscosity ; drug effects ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; Exercise Therapy ; Humans ; Male ; Microfilament Proteins ; genetics ; metabolism ; Muscle Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Swimming

AIMTo investigate the inhibitory effect of egg white lysozyme (LZM) on ceftazidime (CFT)-induced release of endotoxin from Pseudomonas aeruginosa.

METHODSP. aeruginosa PAO1 was inoculated in nutrition broth or diluted rabbit blood free of antibiotics in the presence or absence of LZM and incubated at 37 degrees C on a water bath shaker. beta-Lactam antibiotic, CFT, was added to cultures at 3.5 h (nutrition broth culture) or 5 h (diluted rabbit blood culture) after inoculation. After 3 h of CFT treatment, the supernatants from different bacterial cultures were prepared by centrifuge and the concentrations of endotoxin in the supernatants were measured. The bacterial supernatants were also added to a murine macrophage cell line RAW 264.7 or intravenously injected into carrageenin-sensitized mice. Tumor necrosis factor-alpha (TNF alpha) and nitric oxide (NO) concentrations in RAW 264.7 supernatants or in mouse sera were tested.

RESULTSCFT treatment alone obviously inhibited the growth of P. aeruginosa PAO1 accompanied by strong and rapid bacteriolysis and released relatively high concentration of endotoxin from bacteria both in nutrition broth and in diluted rabbit blood cultures. The bacterial supernatant from CFT treatment alone yielded high concentrations of TNF alpha both in RAW 264.7 cells and in mice and high level of NO in RAW 264.7 cells. Treatment with the combination of LZM and CFT evidently blocked the lysis of bacteria and reduced the release of endotoxin without decreasing bactericidal activity of CFT. TNF alpha and NO productivity of the supernatants prepared from the LZM/CFT combinative treated bacterial cultures were significantly decreased both in RAW 264.7 cells and in mice indicating that the inflammatory activity was reduced.

CONCLUSIONLZM can effectively prevent CFT-induced bacteriolysis, endotoxin release and subsequent proinflammatory factor production but without decreasing bactericidal activity of CFT, resulting in the disassociation of bactericidal activity and bacteriolysis. Thus, LZM might be important for preventing endotoxemia in Gram-negative sepsis with the treatment of antibiotics.

Animals ; Bacteriolysis ; drug effects ; Ceftazidime ; pharmacology ; Egg White ; analysis ; Endotoxins ; metabolism ; Mice ; Muramidase ; isolation & purification ; pharmacology ; Nitric Oxide ; metabolism ; Pseudomonas aeruginosa ; metabolism ; physiology ; Rabbits ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo compare therapeutic effects of traditional moxibustion and moxibustion on temperature-sensitive points for lumbar disc herniation.

METHODSOne hundred and twenty cases were randomly divided into a temperature-sensitive point group and a traditional moxibustion group, 60 cases in each group. The temperature-sensitive point group was treated with moxibustion on the temperature-sensitive points in the temperature-sensitive high incidence area such as waist and lower limbs, once each day; the traditional moxibustion group was treated with warming moxibustion at Jiaji (EX-B 2), Ciliao (BL 32), Zhibian (BL 54), Huantiao (GB 30), Weizhong (BL 40), Yanglingquan (GB 34), Kunlun (BI. 60), once each day, 7 days constituting one course. The therapeutic effect and the recurrence rate were observed after one course treatment and six months later.

RESULTSAfter treatment for one course, the cured-markedly effective rate was 65.00% in the temperature-sensitive point group, superior to 50.0% in the traditional moxibustion group, with a significant difference between the two groups (P < 0.05); all effective cases were followed-up for six months, the cured-markedly effective rate and the recurrence rate were 62.3% and 26.4% in the temperature-sensitive point group, and 34.2% and 46.3% in the traditional moxibustion group, the therapeutic effect of the temperature-sensitive point group being superior to the traditional moxibustion group, and the recurrence rate was lower than the traditional moxibustion group (both P < 0.05).

CONCLUSIONMoxibustion on temperature-sensitive points is a effective therapy for lumbar disc herniation, with stable therapeutic effect and low recurrence rate.

Acupuncture Points ; Adult ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc Displacement ; therapy ; Lumbar Vertebrae ; Male ; Middle Aged ; Moxibustion ; methods ; Temperature ; Treatment Outcome ; Young Adult

OBJECTIVEThe aim of this study was to investigate the protective effect of Chuanxiong-pathalide A on the injury of endothelial cell induced by ischemia and reperfusion in isolated rat hearts.

METHODThe isolated rat hearts were perfused under constant pressure with Chuanxiong-pathalide A at the concentrations of 0.012 5 mg x mL(-1), 0.025 mg x mL(-1) and 0.05 mg x mL(-1) within 10 min followed by a 10-min washout period before the induction of a 30-min normothermic global ischemia and 60 min reperfusion.

RESULTPretreatment with Chuanxiong-pathalide A produced a reduction in the incidence of reperfusion-induced ventricular fibrillation (VF) and ventricular tachycardia (VT). Pretreatment of the hearts with high dose of Chuanxiong-pathalide A (0.05 mg x mL(-1)) prior to the ischemia and reperfusion, reduced the incidence of reperfusion-induced VF and VT to 37.5% as compared with non-pretreated control group (P < 0.05). The duration of occurrence of VF and VT in the group pretreated with Chuanxiong-pathalide A at dosages of 0.012 5 mg x mL(-1), 0.025 mg x mL(-1) and 0.05 mg x mL(-1) were (7.50 +/- 1.61), (1.64 +/- 0.67) and (1.06 +/- 0.70) min respectively, which were significantly shorter than (23.51 +/- 3.99) min in non-pretreated control group; 0.05 mg x mL(-1) of Chuanxiong-pathalide A increased coronary flow in comparison with the control group. The content of SOD in the group pretreated with 0.05 mg x mL(-1) of Chuanxiong-pathalide A was (59.6 +/- 18.7) U x mg(-1), significant lower than (92.3 +/- 19.0) U x mg(-1) in the non-pretreated group. The contents of LDH and MDA were (2 378.0 +/- 196.3) U x g(-1) and (12.1 +/- 1.3) nmol x mg(-1) in the non-pretreated group, which were much higher than the values of (1 669.4 +/- 192.5) U x g(-1) and (6.9 +/- 0.8) nmol x mg(-1) in the group pretreated with Chuanxiong-pathalide A respectively. In addition, enzymeimmunoassays showed an decreased production of IL-1beta and the ratio of TXB2/6-Keto-PGF1alpha.

CONCLUSIONOur results show that chuanxiong-pathalide A pretreatment can protect the endothelial function from the injury caused by ischemia and reperfusion.

Animals ; Benzofurans ; isolation & purification ; pharmacology ; Coronary Vessels ; drug effects ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Endothelial Cells ; drug effects ; pathology ; In Vitro Techniques ; Interleukin-1beta ; blood ; Ischemic Preconditioning, Myocardial ; L-Lactate Dehydrogenase ; metabolism ; Ligusticum ; chemistry ; Malondialdehyde ; metabolism ; Myocardial Reperfusion Injury ; complications ; metabolism ; physiopathology ; Myocardium ; metabolism ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Regional Blood Flow ; drug effects ; Tachycardia, Ventricular ; prevention & control ; Ventricular Fibrillation ; prevention & control

OBJECTIVETo observe the antioxidant effects of water extract of Carthamus tinctorius on ox-LDL induced injury in rat cardiac microvascular endothelial cell and detecting oxygen derived free radicals (OFR) to explore the antioxidant mechanisms.

METHODBy using the third generation of rat cardiac microvascular endothelial cells (rCMEC), the protection of water extract of C. tinctorius was investigated after ox-LDL (100 mg x L(-1) induced damage. The supernatant was collected for detecting lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), xanthine oxidase (XOD), glutathione peroxidase (GSH-Px), nitric oxide (NO), nitric oxide synthase (NOS) activity, and cell suspension was collected for detecting reactive oxygen species (ROS) by electron spin resonance (ESR).

RESULTWater extract of C. tinctorius increased the rCMEC survival rate, reduced LDH, MDA and XOD levels, and improved SOD, GSH-Px and NOS activity, while in the cell suspension ROS signal decreased significantly.

CONCLUSIONWater extract of C. tinctorius has antioxidation. The mechanisms are likely related with scavenging of free radicals, enhancing its clearance, enhancing endogenous antioxidant activity.

Animals ; Carthamus tinctorius ; chemistry ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Electron Spin Resonance Spectroscopy ; methods ; Endothelial Cells ; drug effects ; metabolism ; Enzyme Activation ; drug effects ; Glutathione Peroxidase ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Lipoproteins, LDL ; pharmacology ; Microvessels ; cytology ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Water ; chemistry ; Xanthine Oxidase ; metabolism

OBJECTIVETo observe the protective effects of catechin morphon (GCG and EGCG) on hypoxia-reoxygenation induced injury in myocardial cells and to explore the mechanisms.

METHODIn cultured neonatal rat cardiomyocytes, we investigated the preconditioning protection by GCG and EGCG on the spontaneous beating, the survival rate, the release of LDH, MDA, SOD, GSH-Px and the ATP enzyme activity of cardiomyocyte cellular membrane in cultured rat cardiomyocytes treated during the reoxygenation 1h following hypoxia 3 h. The blocking agent of protien kinase C staurosporine (10 nmol x L(-1)) or the deactivator of Gi/o protein pertussis toxin (PTX, 200 microg x mL(-1)) were added before the catechin treatment.

RESULTSPreconditioning by GCG and EGCG increased the spontaneous beating and the survival rate, and decreased the release of LDH and MDA with the rise of SOD and ATP enzyme activity. Inhibition of PKC by staurosporine and Gi/o protein by PTX abolished the protection by catechin with the reduction of the beating, survival rate and activity of SOD, and the increase of the release of LDH and MDA. The results indicated that the activation of signal transduction pathway from PKC and Gi/o protein seemed to be involved in the cardioprotection of preconditioning by GCG and EGCG.

CONCLUSIONThe protection by GCG and EGCG on hypoxia-reoxygenation injury in cultured neonatal rat cardiomyocytes is found, which is related with scavenging of free radicals, and PKC Gi/o signal transduction pathway.

Animals ; Cardiotonic Agents ; chemistry ; pharmacology ; Catechin ; analogs & derivatives ; chemistry ; pharmacology ; Female ; Free Radicals ; metabolism ; Hypoxia ; complications ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Oxygen ; metabolism ; Protein Kinase C ; metabolism ; Proteins ; metabolism ; Rats ; Signal Transduction ; drug effects ; Survival Rate

OBJECTIVETo study the protected effect of Yindan Xinnaotong capsule (YDXNTC) and main components compatibility on myocardium ischemia/reperfusion injury.

METHODGlobal ischemia/reperfusion was adopted to induce myocardial ischemia/reperfusion injury (MIRI) in isolated rat heart. Sprague-Dawley (SD) rats were divided into control, model, YDXNTC, Ginkgo biloba extract (GBE) group, ethanol extract of Salvia miltiorrhiza (SM-E) group, aqueous extract of Salvia miltiorrhiza (SM-H) group, mixed compatibility of other components in YDXNTC (MC), GBE and SM-E compatibility (GSEC), GBE and SM-H compatibility (GSHC), and SM-E and SM-H compatibility (SEHC). During the experiment, electrocardiogram was recorded to observe cardiac arrest time, heart resuscitation time, regaining normal rhythm time, the incidence and duration of arrhythmias (VT/VF). At the end of reperfusion, hearts were arrested and homogenated to assay the activity of superoxide dismutase (SOD), and the content of malondialdehyde (MDA), lactate dehydrogenase (LDH), creatine kinase-MB (CK-MB), cardiac troponin I.

RESULT(1) YDXNTC, SM-E, SM-H and MC elevated cardiac arrest time, also reduced rebeating time, restoring normal rhythm time as well as the duration of arrhythmia, but no remarkable impact on VT/VF occurrence. GBE was effective for incidence of VT/VF, also achieved good effect on shortening rebeating time, restoring normal rhythm time and arrhythmia duration. Likewise, obviously reduced rebeating time, restoring normal rhythm time and arrhythmia duration, and evaluated cardiac arrest time were also exhibited in compatibility groups except that no lengthened cardiac arrest time was detected in GSHC. And the incidence of VT/VF was decreased by GSEC. (2) YDXNT, ginkgo biloba extract (GBE), ethanol extract of salvia miltiorrhiza (SM-E), GBE and SM-E compatibility (GSEC), and SM-E and aqueous extract of salvia miltiorrhiza (SM-H) compatibility (SEHC) could improved SOD and decreased MDA level SM-H, mixed compatibility of other elements in YDXNTC (MC) and GBE and SM-H compatibility (GSHC) showed a role on MDA reduction. (3) LDH was declined by YDXNT and SM-H. CK-MB was reduced by GBE, SM-E, SM-H, and GSEC. (4) The release of cTnI was only inhibited by GSEC.

CONCLUSIONYDXNTC, primary materials and main components compatibility has a certain protection effect on MIRI, its mechanism may be related to antioxidant and calcium overload reduction.

Animals ; Arrhythmias, Cardiac ; physiopathology ; prevention & control ; Capsules ; Creatine Kinase, MB Form ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Electrocardiography ; Ginkgo biloba ; chemistry ; Heart ; drug effects ; physiopathology ; In Vitro Techniques ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Myocardial Reperfusion Injury ; metabolism ; physiopathology ; prevention & control ; Myocardium ; metabolism ; pathology ; Phytotherapy ; Plant Extracts ; pharmacology ; Protective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry ; Superoxide Dismutase ; metabolism ; Troponin I ; metabolism

OBJECTIVETo establish a LC-MS/MS method for determining the concentration of tanshinone IIA, salvianolic acid B and paeoniflorin of refined coronary cataplasm in rabbit plasma, in order to determine the concentration of the three main ingredients in blood after transdermal administration and calculate their pharmacokinetic parameters.

METHODRabbits were given refined coronary cataplasm on the basis of 15 g x kg(-1) by transdermal administration to detect the plasma concentration of the three main ingredients using LC-MS/MS. Winnonlin software was used to calculate their major pharmacokinetic parameters.

RESULTTanshinone IIA, salvianolic acid B and paeoniflorin showed good linearity (r>0.999) at 1-100, 50-1 000, 10-1 000 microg x L(-1) respectively in plasma, with average recovery rate of 96.57%, 91.90%, 95.93%, respectively. The RSD within day were less than 15%. After transdermal administration of refined coronary cataplasm in rabbits, the main pharmacokinetic parameters of tanshinone IIA, salvianolic acid B or paeoniflorin were as follows: Cmax (20.85 +/- 12.68), (636.25 +/- 386.91), (787.80 +/- 395.64) microg x L(-1); Tmax (0.49 +/- 0.28), (0.44 +/- 0.27), (0.46 +/- 0.30) h.

CONCLUSIONThe LC-MS/MS method is highly selective and sensitive to determine the concentration of samples in rabbit plasma. The pharmacokinetic characteristics of tanshinone IIA, salvianolic acid B and paeoniflorin are suitable to assess the percutaneous absorption of refined coronary cataplasm.

Animals ; Benzoates ; administration & dosage ; pharmacokinetics ; Benzofurans ; administration & dosage ; pharmacokinetics ; Bridged-Ring Compounds ; administration & dosage ; pharmacokinetics ; Chromatography, Liquid ; methods ; Coronary Disease ; drug therapy ; Diterpenes, Abietane ; administration & dosage ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Glucosides ; administration & dosage ; pharmacokinetics ; Humans ; Male ; Monoterpenes ; Rabbits ; Tandem Mass Spectrometry ; methods