Chan-su (toad venom) has been used as an analgesic agent in China from ancient to modern times. Bufalin, a non-peptide toxin extracted from toad venom, is considered as one of the analgesic components. The molecular mechanism underlying the anti-nociceptive effects of bufalin remains unclear so far. In this study, we investigated the pharmacological effects of bufalin on pain-related ion channels as well as animal models through patch clamping, calcium imaging and animal behavior observation. Using the whole-cell recording, bufalin caused remarkable suppressive effect on the peak currents of Nav channels (voltage gated sodium channels, VGSCs) of dorsal root ganglion neuroblastomas (ND7-23 cell) in a dose-dependent manner. Bufalin facilitated the voltage-dependent activation and induced a negative shift on the fast inactivation of VGSCs. The recovery kinetics of VGSCs were significantly slowed and the recovery proportion were reduced after administering bufalin. However, bufalin prompted no significant effect not only on Kv4.2, Kv4.3 and BK channels heterologously expressed in HEK293T cells, but also on the capsaicin and allyl isothiocyanate induced Ca2+ influx. What's more, bufalin could observably relieve formalin-induced spontaneous flinching and licking response as well as carrageenan-induced thermal and mechanical hyperalgesia in dose-dependent manner in agreement with the results of in vitro experiments. The present results imply that the remarkable anti-nociceptive effects produced by bufalin are probably ascribed to its specific regulation on Nav channels. Bufalin inhibits the Nav channels in a dose-dependent manner, which will provide references for the optimal dose selection of analgesia drugs.

Due to their pseudo-persistence and their biological activity, pharmaceuticals are emerging contaminants of major concern for the environment. The aim of this review is to provide an updated inventory of the contamination of aquatic environments by 43 drugs representing different classes of pharmaceuticals, such as antibiotics, anti-inflammatory drugs, anti-depressants, sex hormones, lipid regulators and beta-blockers. The data collected is focused on contamination levels reported in marine coastal waters and in waste and river waters flowing into the Mediterranean Sea. The most widely produced/prescribed classes of medicines are compared with the substances most widely searched for in the environment. Ranges of pollution levels according to the type of water body are also presented, to examine the fate in sewage treatment plants and the persistence in the environment of the targeted molecules. Levels of pharmaceuticals ranged from 100 to 10,000 or even 100,000 ng·L-1 in sewage waters, dropping to 1 to 10,000 ng·L-1 in rivers and to not detected to 3000 ng·L-1 in sea water. However, this paper evidences a lack of data for seawater and also for several countries along the southern coast of the Mediterranean Sea. In order to assess the risk for aquatic ecosystems associated with pharmaceuticals, experimental ecotoxicological values obtained using normalized acute and/or chronic bioassays carried out with different trophic levels were collected for each drug. Targeted biological species and associated bioassays are classified on the basis of their sensitivity to each class of compounds. Occurrence and ecotoxicology are then linked by using the Hazard Quotient (HQ) to assess the environmental risk caused by pharmaceuticals in the Mediterranean Basin. Correlations between HQ and frequency of detection of pharmaceuticals highlighted thirteen compounds that are cause for concern in Mediterranean fresh and sea waters, such as 17α-ethinylestradiol, metoprolol, 8 antibiotics and 3 analgesics/anti-inflammatories.

Endogenous opioids activate opioid receptors (ORs) in the enteric nervous system to control intestinal motility and secretion. The mu OR mediates the deleterious side effects of opioid analgesics, including constipation, respiratory depression and addiction. Although the delta OR (DOR) is a promising target for analgesia, the function and regulation of DOR in the colon are poorly understood. This study provides evidence that endogenous opioids activate DOR in myenteric neurons which may regulate colonic motility. The DOR agonists DADLE, Deltorphin II and SNC80 inhibited electrically-evoked contractions and induced neurogenic contractions in the mouse colon. Electrical, chemical and mechanical stimulation of the colon evoked the release of endogenous opioids, which stimulated endocytosis of DOR in the soma and proximal neurites of myenteric neurons of transgenic mice expressing DOR fused to eGFP. In contrast, DOR was not internalized in nerve fibers within the circular muscle. Administration of dextran sulfate sodium induced acute colitis, which was accompanied by DOR endocytosis and an increased density of DOR-positive nerve fibers within the circular muscle. The potency with which SNC80 inhibited neurogenic contractions was significantly enhanced in the inflamed colon. This study demonstrates that DOR-expressing neurons in the mouse colon can be activated by exogenous and endogenous opioids. Activated DOR traffics to endosomes and inhibits neurogenic motility of the colon. DOR signaling is enhanced during intestinal inflammation. This study demonstrates functional expression of DOR by myenteric neurons and supports the therapeutic targeting of DOR in the enteric nervous system.