Yaozong Pan

Typhonii rhizoma, a widely used herb in traditional Chinese medicine, has acute irritating toxicity related to Typhonium giganteum lectin (TGL). TGL exhibits acute inflammatory effects, but the underlying molecular mechanisms are largely unknown. This paper is designed to assess the pro-inflammatory response of TGL on RAW 264.7 cells. RAW 264.7 treated with 6.25, 12.5, 25, and 50 µg/mL TGL showed elevated levels of inflammatory factors (TNF-α, IL-1β) and of p-IκB and p-p65, all dose-dependent, indicating that TGL had a substantial inflammatory effect and mobilized the nuclear factor-κB (NF-κB) pathway. All four TGL treatments also induced the up-regulation of reactive oxygen species (ROS) and cytosolic free Ca²⁺ and down-regulation of mitochondrial membrane potential (MMP). The production of cytokines and p-IκB, p-p65 were reduced by N-acetylcysteine (NAC), an ROS scavenger, which somewhat abrogated ROS production. The results showed the TGL-activated inflammatory signaling pathway NF-κB to be associated with the overproduction of ROS. Moreover, 50 μg/mL treatment with TGL led to cell apoptosis after 1 h and increased necrosis over time. These results provided potential molecular mechanisms for the observed inflammatory response to TGL including up-regulation of ROS and cytosolic free Ca²⁺, down-regulation of MMP, the mobilization of the NF-κB pathway, and the subsequent overproduction of pro-inflammatory factors resulting in apoptosis. Long-term stimulation with TGL resulted in strong toxic effects related to inflammation that induced necrosis in macrophages.

To look for the toxicity fraction of Euphorbia pekinensis and discuss the vinegar processing mechanism. The level of intestinal edema, water content of intestine and stool, IC50 values of IEC-6 were applied to evaluate the toxicity of different fractions. RT-PCR was employed for detecting AQP1, AQP3 mRNA expression. The petroleum ether (PE) fraction and ethyl acetate (EtOAc) fraction could significant cause intestinal edema in mice, increase the water content of duodenum, colon and stool, inhibited the mRNA expression of AQP1 and increased the mRNA level of AQP3 in colon, and the petroleum ether (PE) fraction was more poisonous. After the petroleum ether (PE) fraction was processed with vinegar, the level of intestinal edema, water content of duodenum, colon, stool and inhibition ratio of cells line were reduced. And we compared the composition change after vinegar processing, finding that the conpekinensis.

The Traditional Chinese Medicine herbs Pollen Typhae and Pollen Typhae Carbonisatus have been used as a hemostatic medicine promoting blood clotting for thousands of years. In this study, a reliable, highly sensitive method based on LC-MS/MS has been developed for differentiation of the heating products of total flavonoids in Pollen Typhae (FPT-N). Twenty three peaks were detected and 18 peaks have been structurally identified by comparing retention times, high resolution mass spectrometry data, and fragment ions with those of the reference substances and/or literature data. Additionally, 15 compounds have been quantified by multiple reaction monitoring in the negative ionization mode. It was found that the contents of the characterized compounds differed greatly from each other in FPT-N samples. Among them, the content of huaicarbon B significantly increased at first, while it decreased after heating for 25 min, which could be considered as the characteristic component for distinguishing FPT-N. The present study provided an approach to rapidly distinguish the differences of FPT-N samples. In addition, the actively summarized characteristic fragmentation might help deducing the structure of unknown flavonols compounds. Furthermore, transformation rules of flavonoids during the heating process in carbonisatus development could contribute to hemostatic therapeutic component exploration.

Platycladi cacumen and its processed product have been utilized as a Chinese medicine to treat hemorrhages. In this study, the base peak chromatogram fingerprints of heating products of total flavonoids in Platycladi cacumen were established by high performance liquid chromatography coupled with mass spectroscopy/mass spectroscopy (HPLC-MS/MS), and the hemostatic activities were studied by hemostatic screening tests in vivo. The spectrum-effect relationships between fingerprints and hemostatic activities were analyzed by using canonical correlation analysis to trace the peaks responsible for the significant hemostatic effects. Peak 10 and peak 12 were correlated most closely, thus probably being the main hemostatic compounds. To confirm the reliability of this strategy, the targeted unknown peak was obtained by bioactivity-guided isolation, characterized by MS, ¹H-NMR, ¹³C-NMR, and 2D-NMR spectroscopies, and referred to as cecarbon as a new compound. In addition, the isolated compound exhibited hemostatic effect in a dose-dependent manner with different potencies in vitro and existed in Platycladi cacumen Carbonisatus. A novel dereplication strategy was employed to trace and identify the active compounds of other herbs that have bioactivity enhancement after processing using spectrum–effect relationships and column chromatography.

To explore the antagonistic effect of gingerols against the inflammation induced by lectin from Pinellia ternata. In this study, ELISA method was used to determine the effect of different extracts from gingerols on the release of inflammatory factor TNF-α from macrophages induced by lectin from P. ternata. The fluorescence probe was used to determine the effect of gingerols on the changes in ROS of macrophages induced by lectin from P. ternata. The western-blot method was applied to study the effect of gingerols on the increase in expression of cell receptor interacting protein RIP3 in macrophages induced by lectin from P. ternata. The scanning electron microscope (SEM) was used to study the effect of gingerols on morphological changes in macrophages induced by lectin from P. ternata. According to the results, gingerols can significantly inhibit the release of inflammatory factor from macrophages induced by lectin from P. ternata, ROS overproduction and increase in RIP3 expression. SEM results showed that gingerols can inhibit the cytomorphosis and necrocytosis induced by lectin from P. ternata. Fresh ginger's detoxication may be related to gingerols' effects in inhibiing release of inflammatory factor, ROS overproduction and increase in RIP3 expression caused by macrophages induced by lectin from P. ternata, which are mainly inflammatory development.