ABSTRACT
OBJECTIVE In this study we explored the role of Epac1-Rap1 pathway in the acute myocardial ischemia/reperfusion injury (MIRI) in vitro and in vivo. METHODS An acute myocardial ischemia/reperfusion injury model was established by the ligation of left anterior descending coronary. Myocardial architecture, fibers and apoptosis was evaluated by the Masson trichrome staining, Sirius red staining and TUNEL assay.H9c2 cells were subjected to hypoxia for 5 h followed by 1-h reoxygen-ation in vitro.Cell viability was measured by MTT assay and cellular injury was evaluated by measuring the release of lactate dehydrogenase (LDH). Western blot, real-time PCR and immunofluorescence were used to detect the expressions of Epac1 and relative downstream molecules.RESULTS Myocardial IR-induced cardiac apoptosis and accumulation of Epac1 and Rap1 in rat IR injury model.Direct Epac activation by 8-CPT(8-(4-chlorophenylthio)-2′-O-methyl-cAMP)exacerbated cardiomyocyte death and dysfunction following hypoxia-reoxygenation(H/R),selective activation of Epac in response to H/R was evident which enriched for cytosolic/membrane proteins and mRNA. Harmacological inhibitor of Epac (ESI-09)significantly ameliorated myocardial injury with the decline of Epac expression.Epac inhibitor and agonist studies also implicated the effect of Rap1, which is downstream of Epac in this pathway. The expression of Rap1 elevated when activated by Epac agonist and was blocked by Epac inhibitor. The same result was true for myocyte CaMK-II and intracellular calcium ions activation.Moreover,ESI-09 also increased ERK1/2 phosphorylation. CONCLUSION Our study reveal that Epac1/Rap1 signaling pathway is involved in the pathogenesis of myocardial I/R injury in rats,which provides evidence on the development of therapeutic strategies target this pathway for myocardial I/R injury.
ABSTRACT
In this paper, menstruation prescriptions were selected from "Fu Qingzhu's Obstetrics and Gynecology" and analyzed by using GRI algorithm, correlation analysis, hierarchical clustering method through SPSS, Clementine and traditional Chinese medicine (TCM) inheritance auxiliary systems, in order to screen out 15 menopathy prescriptions, which involve 45 traditional Chinese medicine herbs. In the study, blood-tonifying and qi-tonifying herbs were found to be frequent in the prescriptions. The most frequent single herb was white paeony root, accounting for 9.6% in the total number of prescriptions; The most frequent herb pairs were white paeony root-radix rehmanniae preparata and paeony root-angelica sinensis. Among Fu Shan's menopathy prescriptions, 61 herbal pairs showed a correlation coefficient exceeding 0.05, which evolved into 16 pairs of core combinations. The analysis showed that menopathy prescriptions in volume 1 of "Fu Qingzhu's Obstetrics and Gynecology" focused on tonic traditional Chinese medicines involving liver, spleen and kidney and were adjusted according to changes in qi, blood, cold, hot and wet, which could provide a specific reference for further studies on Fu Shan's academic thoughts and traditional Chinese medicine clinical treatment of menopathy.
Subject(s)
Female , Humans , Books , History , China , Drug Prescriptions , History , Drug Therapy , History , Drugs, Chinese Herbal , Chemistry , Therapeutic Uses , Gynecology , History, Ancient , Medicine in Literature , MenstruationABSTRACT
<p><b>OBJECTIVE</b>To evaluate the effect of stretch on sarco(endo)plasmic reticulum Ca2+-Mg2+ ATPases activity and mRNA level and study the remodeling reaction of muscle in a variety of mechanical environments.</p><p><b>METHODS</b>Myoblast from maxillofacial skeletal muscle of one-week-old male Sprague-Dawley rat was cultured and stretched cyclicly using a four-point bend device. Inorganic Phosphorus test was used to compare the activity of Ca2+-Mg2+ ATPases of myoblast before and after stretch. RT-PCR was also used to observe the Ca2+-Mg2+ ATPases mRNA level.</p><p><b>RESULTS</b>The activity of Ca2+-Mg2+ ATPases of myoblast down regulated significantly in 4 hours. During the period of 8 hours to 24 hours, up-regulation followed then returned to control level at the 48 hour point. RT-PCR showed that Ca2+-Mg2+ ATPases mRNA level were elevated by stretch, particularly at 2 hour and 48 hour point.</p><p><b>CONCLUSION</b>The results suggested a transcriptional control of Ca2+-Mg2+ ATPases activity was involved in the muscle remodeling process induced by stretch.</p>