ABSTRACT
<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>We 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.</p><p><b>RESULTS</b>The 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.</p><p><b>CONCLUSION</b>Evidence 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).</p>
Subject(s)
Animals , Female , Humans , Male , Mice , Middle Aged , Rats , Cell Survival , Genetics , Cytokines , Bodily Secretions , Drugs, Chinese Herbal , Pharmacology , Gene Expression Regulation , Healthy Volunteers , Inflammation Mediators , Metabolism , Inhibitory Concentration 50 , Macrophages , Metabolism , RNA, Messenger , Genetics , Metabolism , Rats, Sprague-Dawley , Syndrome , Toll-Like Receptors , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To 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.</p><p><b>METHOD</b>RAW 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.</p><p><b>RESULT</b>Shensu 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.</p><p><b>CONCLUSION</b>Depressing the expression of TLR3, MyD88, TRAM and TRIF mRNA may be the elementary basis of Shensu Yin to play heat-clearing and detoxicating effect.</p>
Subject(s)
Animals , Male , Mice , Rats , Adaptor Proteins, Vesicular Transport , Genetics , Cell Line , Drug Combinations , Drugs, Chinese Herbal , Pharmacology , Interferon-beta , Bodily Secretions , Lipopolysaccharides , Pharmacology , Macrophages , Cell Biology , Metabolism , Myeloid Differentiation Factor 88 , Genetics , Plants, Medicinal , Chemistry , Poly I-C , Pharmacology , RNA, Messenger , Genetics , Random Allocation , Rats, Sprague-Dawley , Receptors, Interleukin , Genetics , Signal Transduction , Toll-Like Receptor 3 , Genetics , Toll-Like Receptor 4 , Genetics , Tumor Necrosis Factor-alpha , Bodily SecretionsABSTRACT
<p><b>OBJECTIVE</b>To 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.</p><p><b>METHOD</b>rCMEC 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.</p><p><b>RESULT</b>Positive 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.</p><p><b>CONCLUSION</b>2-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.</p>