To further uncover the scientific significance and molecular mechanism of the Chinese herbs with pungent hot or warm natures, endogenous and exogenous expression systems were established by isolation of dorsal root ganglion (DRG) neurons and transfection of HEK293 cells with TRPV1 channel gene separately. On this basis, the regulation action of capsaicin, one main ingredient from chili pepper, on TRPV1 channel was further explored by using confocal microscope. Besides, the three-sites one-unit technique and method were constructed based on the brown adipose tissue (BAT), anal and tail skin temperatures. Then the effect of capsaicin on mouse energy metabolism was evaluated. Both endogenous and exogenous TRPV1 channel could be activated and this action could be specifically blocked by the TRPV1 channel inhibitor capsazepine. Simultaneously, the mice's core body temperature and BAT temperature fall down and then go up, accompanied by the increase of temperature of the mice's tail skin. Promotion of the energy metabolism by activation of TRPV1 channel might be the common way for the pungent-hot (warm) herbs to demonstrate their natures.
Adipose Tissue, Brown ; drug effects ; physiology ; Animals ; Capsaicin ; analogs & derivatives ; pharmacology ; Energy Metabolism ; Ganglia, Spinal ; cytology ; HEK293 Cells ; Humans ; Mice ; Neurons ; drug effects ; physiology ; Plants, Medicinal ; chemistry ; TRPV Cation Channels ; physiology ; Temperature ; Thermogenesis

OBJECTIVETo study the antiinflammatory effect of a compound TCM (Traditional Chinese Medicine) agent on animal models. The agent contains ant extractive and a blent of three herbal products, herba epimedii, fructus cnidii, and fructus lycii.

METHODThree animal models to induce experimental inflammation in rats, including carrageenin--induced paw edema, cotton-ball granuloma and adjuvant induced arthritis, were chosen to study the antiinflammatory effect of the TCM agent.

RESULTThe TCM agent showed a marked inhibitory effect on edema induced by all three types of inflammation in rats, the inhibitory rate of the TCM agent at the dose of 0.20, 0.40 and 0.80 g.kg-1 in granuloma model bing over 25% at 1 hour post oral administration, and being 23.8%, 22.7%, 39.7% at 6 hour. In addition, the TCM agent also showed a significant preventive as well as therapeutic effect on adjuvant induced arthritis in rats, and improved the pathological changes of the animal joints with the induced arthritis.

CONCLUSIONTCM agent has significant antiinflammatory effects on the three above mentioned animal models.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Ants ; Arthritis ; drug therapy ; Capsules ; Cnidium ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Edema ; drug therapy ; Epimedium ; chemistry ; Granuloma, Foreign-Body ; drug therapy ; Lycium ; chemistry ; Male ; Materia Medica ; therapeutic use ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar

OBJECTIVETo investigate the changes of the activity of both protein kinase A and C and the mechanisms of antipyretic action of Guizhi decoction.

METHODThe fever responses were observed after combination injection of H-89 (a selective inhibitor of PKA) and calphostin C (a selective inhibitor of PKC), and oral pretreatment of Guizhi decoction in fever rats induced by an intra-cerebroventricular (icv) injection of an EP3 agonist, and both PKA and PKC activity in hypothalamus were measured in rats pretreated with Guizhi decoction and vehicle using isotopic tracing assay.

RESULTThe rise in rat body temperature was inhibited by H-89, Calphostin C, and Guizhi decoction, moreover, pretreatment with Guizhi decoction reduced PKA activity obviously. PKC activity in model rats exhibited a tendency to drop compared with that of control group, Oral administration of Guizhi decoction in large dose inhibited the response significantly, while the low dose of Guzhi decoction has no effect on PKC.

CONCLUSIONBoth PKA and PKC may participate in the mechanism of fever induction by EP3 agonist. The decrease of PKA and PKC may contribute to the antipyretic action of Guizhi decoction, some isoenzyme of PKC may play a role in the fever production.

Analgesics, Non-Narcotic ; pharmacology ; Animals ; Cinnamomum aromaticum ; chemistry ; Cyclic AMP-Dependent Protein Kinases ; metabolism ; Dinoprostone ; analogs & derivatives ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Fever ; chemically induced ; enzymology ; Hypothalamus ; enzymology ; Male ; Plants, Medicinal ; chemistry ; Protein Kinase C ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, Prostaglandin E ; agonists ; Receptors, Prostaglandin E, EP3 Subtype

OBJECTIVETo investigate the influences of Shensu Yin to RAW 264.7 on the expression of TLR3, TLR4 and the factors of the downstream in RAW 264. 7 cells.

METHODRAW 264.7 cell line was stimulated with Lipopolysaccharide and POLY I: C, respectively, and treated with the drug serum of Shensuyin simultaneously. 24 hours later, collected the supernatant and measured the inflammatory factors TNF-alpha and IFN-beta, extracted mRNA and measured the expression of TLR3, TLR4 and other correlated indexes of the downstream, analyzed and evaluated Shensu Yin's substance basis of pharmacodynamic actions.

RESULTShensu Yin drug serum depressed the expression of TLR4, MyD88, TRAF-6, TRAM and TRIF mRNA, as a result, it decreased the amount of TNF-alpha and IFN-beta.

CONCLUSIONDepressing the expression of TLR3, MyD88, TRAM and TRIF mRNA may be the elementary basis of Shensu Yin to play heat-clearing and detoxicating effect.

Adaptor Proteins, Vesicular Transport ; genetics ; Animals ; Cell Line ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Interferon-beta ; secretion ; Lipopolysaccharides ; pharmacology ; Macrophages ; cytology ; drug effects ; metabolism ; Male ; Mice ; Myeloid Differentiation Factor 88 ; genetics ; Plants, Medicinal ; chemistry ; Poly I-C ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Interleukin ; genetics ; Signal Transduction ; drug effects ; Toll-Like Receptor 3 ; genetics ; Toll-Like Receptor 4 ; genetics ; Tumor Necrosis Factor-alpha ; secretion

To observe the effects of phenylallyl compounds on prostaglandin E2 (PGE2) release in mouse cerebral microvascular endothelial cells (bEnd. 3) stimulated by IL-1beta, and to analyze their structure-activity relationship. Different concentrations of phenylallyl compounds were added separately, and the content of PGE2 induced by IL-1beta in the culture media was measured by ELISA assay. The 50% inhibitory concentration (IC50) of PGE2 was calculated. Studies showed that phenylallyl compounds could affect the PGE2 release differently in bEnd. 3 cells induced by IL-1beta. Close relationships were shown between the inhibitory activities and the location and number of the substituent groups. In conclusion, phenylallyl compounds exhibited inhibitory activities at different extent on PGE2 release in bEnd. 3 cells stimulated by IL-1beta and presented certain structure-activity relationship.
Acrolein ; analogs & derivatives ; isolation & purification ; pharmacology ; Animals ; Brain ; blood supply ; Cells, Cultured ; Cinnamates ; isolation & purification ; pharmacology ; Dinoprostone ; antagonists & inhibitors ; secretion ; Drugs, Chinese Herbal ; chemistry ; Endothelial Cells ; cytology ; metabolism ; Inhibitory Concentration 50 ; Interleukin-1beta ; pharmacology ; Mice ; Microvessels ; cytology ; Propanols ; isolation & purification ; pharmacology ; Structure-Activity Relationship

OBJECTIVETo investigate the effect of Guizhi Tang and its active components on the fever induced by EP3 receptor agonist sulprostone in rats.

METHODThe rise in body temperature evoked by a LCV(lateral cerebroventricle)-injection of sulprostone was compared with that of sulprostone induced-fever rats pretreated with Guizgi Tang and its active compounds, cinnamaldehyde, cinnamic acid and total glucosides of paeony.

RESULTPretreatments with Guizhi Tang and cinnamaldehyde inhibited the rise in body temperature induced by sulprostone, while cinnamic acid tended to augment the fever. The sulprostone-induced fever was blocked by an ip pretreatment of total glucosides of paeony even below the basement.

CONCLUSIONPresent data suggest that interruption with the down-stream events of EP3 receptor may contribute to the antipyretic action of Guizhi Tang, cinnamaldehyde and the total glucosides of paeony, while cinnamic acid may have no such effect.

Acrolein ; analogs & derivatives ; isolation & purification ; pharmacology ; Analgesics, Non-Narcotic ; isolation & purification ; pharmacology ; Animals ; Body Temperature ; drug effects ; Cinnamates ; isolation & purification ; pharmacology ; Dinoprostone ; analogs & derivatives ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Fever ; chemically induced ; physiopathology ; Glucosides ; isolation & purification ; pharmacology ; Male ; Paeonia ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Receptors, Prostaglandin E ; agonists ; Receptors, Prostaglandin E, EP3 Subtype

OBJECTIVETo observe the effect of 2-methoxycinnamaldehyde (isolated from fraction A of Guizhi Tang) on activity of COX and PGE2 release in rat cerebral microvascular endothelial cells (rCMEC) stimulated by IL-1.

METHODrCMEC were cultured, and identified by immunohistochemistry for von Willebrand factor (VIII factor, a marker for all endothelial cells) in cytoplasm of the cells. Different concentrations of 2-methoxycinnamaldehyde were added respectively and incubated for 3 hours, then stimulated for another 12 hours by IL-1. Activities of COX-1 and COX-2 in rCMEC, and production of PGE2 in the conditioned media were measured by ELISA.

RESULTPositive immunostaining for VIII factor was present diffusely in the cytoplasm of > 90% rCMEC. After being exposed to 30 ng x mL(-1) IL, the activity of COX-2 in rCMEC and the production of PGE2 in conditioned media were higher than those of control group, while there was no difference on activity of COX-1 in the two groups. 2-methoxycinnamaldehyde could down-regulate them in concentration-dependently, and significant differences on the activity of COX-2 and amount of PGE2 were showed in 200 microg x mL(-1) concentration.

CONCLUSION2-methoxycinnamaldehyde can affect the PGE2 release in rCMEC induced by IL-1, which might be related with its inhibition on the activity of COX-2.

Acrolein ; administration & dosage ; analogs & derivatives ; isolation & purification ; pharmacology ; Animals ; Brain ; blood supply ; Cells, Cultured ; Cyclooxygenase 1 ; metabolism ; Cyclooxygenase 2 ; metabolism ; Dinoprostone ; metabolism ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Endothelial Cells ; cytology ; metabolism ; Interleukin-1 ; antagonists & inhibitors ; Male ; Microcirculation ; cytology ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the pathological mechanisms of Guizhi Decoction () syndrome and the therapeutic molecular mechanisms of the Guizhi Decoction, Mahuang Decoction (), Sangju Decoction ( ) and Yinqiao Powder (), as well as the potentially biological basis that Guizhi Decoction is most effective only for the patients with Guizhi Decoction syndrome in clinical practice.

METHODSWe first got serum samples from the patients suffering from both upper respiratory tract infection and Guizhi Decoction syndrome identified by the doctors of Chinese medicine (CM) in the clinic. Four formulas with therapeutic actions of pungent warmth or pungent coolness for superficial syndromes were chosen and four kinds of rat serum samples each containing one of the above-mentioned herbal formulas were collected, then the effects of Guizhi Decoction syndromes' patient serum as well as the effects of sera containing the formulas after being stimulated by the patient serum samples on both the mRNA expression of certain toll-like receptor (TLR) subtypes and the release of some inflammatory cytokines in RAW264.7 cells were tested and analyzed in vitro.

RESULTSThe expression of TLR-3, TLR-4 and TLR-9 mRNA among the 9 tested TLR subforms were up-regulated in the macrophages stimulated by the sera from untreated upper respiratory infection patients with the Guizhi Decoction syndrome (symptomcomplex). The products such as interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α and interferon (IFN)-β from stimulated macrophages through TLR signaling pathways were also increased correspondingly. Interestingly, the changes induced by the Guizhi Decoction syndrome patients' sera were masked significantly after the macrophages were incubated with the sera from donors treated with Guizhi Decoction. Similarly, the three other exterior-releasing formulas were all effective in reversing the up-regulated changes of certain TLR subforms to different degrees, but both the number of targeted TLRs and efficacy of them seemed to be inferior to that of Guizhi Decoction.

CONCLUSIONEvidence from these experiments might contribute to the scientific explanation of both the pharmacological mechanisms of Guizhi Decoction and also the CM theory that Guizhi Decoction is specifically prescribed for the treatment of Guizhi Decoction syndrome (The gearing formula to the symptom-complex).

Animals ; Cell Survival ; drug effects ; genetics ; Cytokines ; secretion ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression Regulation ; drug effects ; Healthy Volunteers ; Humans ; Inflammation Mediators ; metabolism ; Inhibitory Concentration 50 ; Macrophages ; drug effects ; metabolism ; Male ; Mice ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Syndrome ; Toll-Like Receptors ; genetics ; metabolism

Objective To investigate the resistant mechanism of quinolones on multi-drug resistant Klebsiella caused pneumonia(MDR-KPN).Methods From August 2008 to May 2010,47 strains of MDR-KPN were collected from 6 hospitals in Hangzhou and Huzhou in Zhejiang province in China.Drug target genes to quinolones (gyrA,parC) and quinolone-resistance genes mediated by mobile genetic elements [qnrA,qnrB,qnrS,aac (6')-Ⅰ b-cr,qepA] were analyzed by PCR and verified by DNA sequencing.Results Positive results were found in 47 strains of MDR-KPN,43 strains (91.5％) of gyrA mutation,40 strains(85.1％) ofparC mutation,3 strains (6.4％) of qnrB2,1 strain (2.1％) ofqnrB 4,8 strains (17.0％) ofqnrS 1,5 strains (10.6％) of qnrS 4,2 strains (4.3％)of aac (6')-Ⅰ b-cr respectively.Moreover,5 novel variants of gyrA (GenBank accession number:JN811952,JN811953,JN811954,JN811955,JN811956),5 novel variants of parC (GenBank accession number:JN817432,JN817433,JN817434,JN817435,JN817436)were also identified.In addition,qnrS4 (GenBank accession number:JN836269) appeared to be the novel variants of qnrS.Conclusion Quinolone-resistance-determining region played a key role on the resistance to quinolones in this group of MDR-KPN,and quinolone-resistance genes mediated by mobile genetic elements [qnrB2,qnrB4,qnrS1,qnrS4,aac (6')-Ⅰ b-cr] showed positive in some parts of the strains.This was the first report on emergence of qnrS4 in the world.

Huanshao capsule is widely used in irregular menstruation and has achieved a good effect. Huanshao capsule can promote gonad development in mice, significantly improve the ovarian index in mice, increase estrogen level and reduce FSH level in rats, inhibit the pain response induced by oxytocin and estrogen, inhibit writhing reaction induced by acetic acid pain in mice. Due to the complexity of traditional Chinese medical formula, the pharmacological mechanism of the treatment on the irregular menstruation of the Huanshao capsule is unclear. In this study, the internet-based computation platform (www.tcmip.cn）was used to explore the molecular mechanism of Huanshao capsule on the menstrual. The aim of this study was to find the molecular mechanism of Huanshao capsule in treating menstrual. In the study of the molecular mechanism of Huanshao capsule in the treatment of menstrual by using the internet-based computation platform, Huanshao capsule maybe treat the menstrual by the pathway of endocrine system, GnRH signal transduction pathway, estrogen signal transduction pathway, oxytocin signaling pathway, thyroid hormone signaling pathway, VEGF signaling pathway, FCεRI signaling pathway and purine metabolism and nucleotide metabolism. The early pharmacological study confirmed Huanshao capsule could increase the serum estradiol level and decrease follicle stimulating hormone level and the traditional Chinese medicine pharmacology coincide with the prediction result of internet-based computation platform which roles as the pathway of GnRH signaling pathway and estrogen signal transduction pathway. Other pathway needs further experimental verification.