Necrostatin-1 attenuates Caspase-1-dependent pyroptosis induced by the RIPK1/ZBP1 pathway in ventilator-induced lung injury.

BACKGROUND: Ventilator-induced lung injury (VILI) is a complex pathophysiological process leading to acute respiratory distress syndrome (ARDS) and poor outcomes in affected patients. As a form of programmed cell death, pyroptosis is proposed to play an important role in the development of ARDS. Here we investigated whether treating mice with the specific RIPK1 inhibitor Necrostatin-1 (Nec-1) before mechanical ventilation could inhibit pyroptosis and alleviate lung injury in a mouse model. METHODOLOGYS: Anesthetized C57BL/6J mice received a transtracheal injection of Nec-1 (5 mg/kg) or vehicle (DMSO) 30 min before the experiment which was ventilated for up to 4 h. Lung damage was assessed macroscopically and histologically with oedema measured as the wet/dry ratio of lung tissues. The release of inflammatory mediators into bronchoalveolar lavage fluid (BALF) was assessed by ELISA measurements of TNF-α,interleukin-1ß (IL-1ß), and IL-6. The expression of RIPK1, ZBP1, caspase-1, and activated (cleaved) caspase-1 were analyzed using western blot and immunohistochemistry, and the levels of gasdermin-D (GSDMD) and IL-1ß were analyzed by immunofluorescence staining. RESULTS: High tidal ventilation produced time-dependent inflammation and lung injury in mice which could be significantly reduced by pretreatment with Nec-1. Notably, Nec-1 reduced the expression of key pyroptosis mediator proteins in lung tissues exposed to mechanical ventilation, including caspase-1, cleaved caspase-1, and GSDMD together with inhibiting the release of inflammatory cytokines. CONCLUSION: Nec-1 pretreatment alleviates pulmonary inflammatory responses and protects the lung from mechanical ventilation damage. The beneficial effects were mediated at least in part by inhibiting caspase-1-dependent pyroptosis through the RIPK1/ZBP1 pathway.

An emission inventory of marine vessels in Shanghai in 2003.

We developed an air pollutant emission inventory for marine vessels in the Shanghai Port in 2003. We estimated emissions under cruising and maneuvering conditions based on two categories of vessels: (1) vessels in the Outer Port, which enter the area following notification of the Shanghai Maritime Safety Administration, a division of the Ministry of Communications of P.R. China; and (2) vessels in internal waterways, which enter those waters following notification of the local Port Administration Centers. Vessels in the Outer Port consist of three subcategories: (1) international vessels that are engaged in foreign commerce; (2) domestic vessels traveling along the downstream portion of the Huangpu River; and (3) domestic vessels traveling along the coast. We also estimate emissions from vessels over 1000 DWT operating under hotelling conditions in the Outer Port. In 2003, the total number of calls was approximately 1.3 million, of which 57% is attributed to vessels in internal waterways and 43% to vessels in the Outer Port. Total marine emissions for NOx, SO2, PM, HC, and CO2 in 2003 are estimated to be 58,160, 51,180, 6960, 4560, and 3,012,800 tons, respectively. Emissions are allocated to 1 km x 1 km grid cells for the 129 km x 102 km Shanghai Port study domain.

Involvement of neuropeptide Y and Y1 receptor in antinociception in the arcuate nucleus of hypothalamus, an immunohistochemical and pharmacological study in intact rats and rats with inflammation.

Neuropeptide Y (NPY) plays an important role in pain modulation at different levels in the central nervous system. In the brain, NPY and NPY receptors distribute abundantly in the arcuate nucleus of hypothalamus (ARC), a structure involved in pain processing. The present study was undertaken to investigate the role of NPY in nociceptive modulation in the ARC of intact rats and rats with carrageenan-induced inflammation. Intra-ARC administration of NPY induced dose-dependent increases in hindpaw withdrawal latencies (HWLs) to thermal and mechanical stimulation in intact rats, which was attenuated by the Y1 receptor antagonist NPY28-36. Intra-ARC administration of NPY also induced dose-dependent increases in HWLs to noxious stimulation in rats with inflammation. Furthermore, intra-ARC injection of either the antiserum against NPY or NPY28-36 induced decreases in HWLs in rats with inflammation, while both of them produced no effects in intact ones. Additionally, there were marked increases of Y1 receptor in the bilateral ARC of rats with inflammation tested by immunohistochemistry, while no significant changes of NPY were observed, implicating that the increased Y1 receptor has an important effect in the NPY-induced antinociception. We also found that intra-ARC injection of Y2 receptor agonist NPY3-36 produced no significant antinociception in either intact rats or rats with inflammation. Together, we demonstrate that NPY exerts an antinociceptive effect in the ARC of intact rats and rats with inflammation. Both Y1 receptor and endogenous released NPY in the ARC are involved in the nociceptive modulation during inflammation.

Plastic changes of calcitonin gene-related peptide in morphine tolerance: behavioral and immunohistochemical study in rats.

The present study was undertaken to investigate the plasticity of calcitonin gene-related peptide (CGRP) in antinociception after morphine tolerance in rats. The hindpaw withdrawal latencies (HWLs) to both thermal and mechanical stimulation increased significantly after intracerebroventricular injection of 2.5 nmol of CGRP in opioid-naive rats, indicating that CGRP produces an antinociceptive effect in the brain. Furthermore, there was an antinociceptive effect after intracerebroventricular injection of 2.5 nmol of CGRP in morphine-tolerant rats. Interestingly, the antinociceptive effect induced by intracerebroventricular injection of CGRP was lower in morphine-tolerant rats than that in opioid-naive rats at the same dose. At the same time, there was downregulation of CGRP-like immunoreactivity in both lateral septal nucleus and central nucleus of amygdala tested by immunohistochemical methods, whereas no significant changes were observed in arcuate nucleus of hypothalamus and periaqueductal gray after morphine treatment in rats. The present study demonstrates plastic changes in both CGRP-induced antinociception and CGRP-like immunoreactivity in rat brain after morphine tolerance, suggesting that CGRP may play an important role in morphine tolerance.

Anti-nociceptive effect of neuropeptide Y in the nucleus accumbens of rats: an involvement of opioid receptors in the effect.

The present study investigated the effect of neuropeptide Y on nociception in the nucleus accumbens of rats. Intra-nucleus accumbens administration of neuropeptide Y induced dose-dependent increases in the hindpaw withdrawal latency (HWL) to thermal and mechanical stimulation in rats. There were no significant changes in the HWL to both stimulation during 60 min after the administration of NPY to outside of the nucleus accumbens. The anti-nociceptive effect of NPY was blocked by subsequent intra-nucleus accumbens injection of the Y1 receptor antagonist neuropeptide Y 28-36, indicating that Y1 receptor is involved in the neuropeptide Y-induced anti-nociception in the nucleus accumbens. Furthermore, the anti-nociceptive effect of neuropeptide Y was attenuated by intra-nucleus accumbens administration of the opioid antagonist naloxone, suggesting an involvement of the endogenous opioid system in the neuropeptide Y-induced anti-nociception in the nucleus accumbens of rats. Moreover, the neuropeptide Y-induced anti-nociception was attenuated by following intra-nucleus accumbens injection of the selective opioid antagonists nor-binaltorphimine and beta-funaltrexamine, but not by naltrindole, illustrating that mu- and kappa-opioid receptors, not the delta-opioid receptor, were involved in the neuropeptide Y-induced anti-nociception in the nucleus accumbens of rats.