Study On The Mechanism Of Gingerols Antagonize The Inflammatory Effect Of Pinella Pedatisecta And Pinellia Ternata

Pinellia ternata (Thunb.) Breit and Pinellia pedatisecta Schott, which belong to poisonous Chinese medicine. Oral intake of the P. ternate or P. pedatisecta will lead to pain and inflammation of the lips, throat, mucosal edema, and even cause death. The toxicity of P. ternate mainly shows as intensive stimulation of mucosal. The results indicated that the irritation toxicity of P. ternate is an inflammatory effect. P. ternate contains needle-like raphides which were named as toxic raphides. Toxic raphides directly stimulate the mucosa, causing severe mucosal damage and release of a large number of inflammatory mediators, causing stimulate pain and inflammatory reaction. Toxic raphides is mainly composed of calcium oxalate and more than 6% proteins. Pinellia ternata lectin (PTL) exhibits acute inflammation effects in vivo. Such pro-inflammatory effects present as massive neutrophils recruitment.Ginger was used to detoxificate to poisonous Chinese medicine of Araceae in clinic. Traditional Chinese medicine theory held that "P. ternata feared ginger, and cooked with ginger for detoxification". Ge Hong held that "P. ternata was toxic and was detoxificated by ginger juice and dry ginger". The traditional Chinese medicine theory may correlated with the anti-inflammatory effects of ginger against toxic raphides and PTL of P. ternate. However, there aren’t such reports so far.This thesis is a further study on the pro-inflammatory mechanism of P. ternate and P. pedatisecta. Meanwhile, the anti-inflammatory mechanism against P. ternate and P. pedatisecta of Gingerols extracted from ginger was well studied to reveal the depth meaning of traditional Chinese medicine theory. Main work and results contain following parts:1 Study on the antagonism of gingerols against inflammatory effect of P. ternate and P. pedatisecta.The mouse peritonitis an toe swelling of rat induced by the raphides and lectinwere applied to investigate the the antagonism of gingerols against inflammatory effect. The result showed that gingerols can decrease the release of inflammatory mediators (PGE2、TNF-a and IL-1β) and toe swelling of rat.SEM was used to observed the morphology changes on the gingerol’s inflyence against P. ternate and P. pedatisecta; Set up of macrophage culture model and study the gingerol’s inflyence on pro-inflammatory factors release; Set up of macrophage-neutrophils co-culture model and neutrophils migration experiment was used to examine the gingerol influence on neutrophils migration against P. ternate and P. pedatisecta stimulation on macrophages; Results showed that the pro-inflammatory effects of P. ternate and P. pedatisecta were caused by activating macrophages, followed by pro-inflammatory factors release, neutrophils migration to inflammation sites and severe inflammation as a result. Moreover, the antagonism of gingerols was to suppress the inflammatory effect caused by P. ternate and P. pedatisecta’s stimulation on macrophages involving the suppression of pro-inflammatory factors release.macrophages activation,neutrophils migration.2 Antagonism mechanism of gingerols against inflammatory effect of P. ternate and P. pedatisectaWestern Blot was used to evaluate the gingerol’s inflyence on expression of NF-κB after PTL and PPL stimulation on macrophages; Use inverted fluorescence microscope and fluorescence microplate to examine the level of ROS released in oxidative stress; Use flow cytometry to study the gingerol’s inflyence on the mitochondrial membrane potential(MMP) and of macrophages stimulated by P. ternate and P. pedatisecta. The results showed that after gingerols stimulation, the level of P65 in cytoplasm significantly raised and P65 in nuclei decreased apparently; ROS release and intracelluar Ca2+ level also decreased in a dose-dependent manner; and the MMP of macrophages significantly raised too.The researches above suggested that the anti-inflammatory effect of gingerol was suppression of the pro-inflammatory effect of P. ternate and P. pedatisecta, involving the suppression of pro-inflammatory factors release, macrophages activation, and neutrophils migration. The anti-inflammatory mechanism of gingerols was to suppressthe inflammatory effect caused by P. ternate and P. pedatisecta’s stimulation on macrophages, involving the suppression of NF-κB and oxidative stress activation, decrease of MMP, and intracelluar Ca2+ leve rising.This thesis is the study on the scientific connotation of the traditional Chinese medicine theory "Ginger detoxificate to the toxicity of poisonous medicines from Araceae". The results thoroughly revealed the pro-inflammatory mechanism of P. ternate and P. pedatisecta’s,and the detoxification mechanism of gingerols, and further confirmed the traditional Chinese medicine theory. The study of this thesis could provide both theoretical basis to Araceae familys’ processing and clinical use, and reference experiment model to the modernization of traditional Chinese medicine.