Induction of Acute Ischemic Stroke in Mice Using the Distal Middle Artery Occlusion Technique

Ischemic stroke remains the predominant cause of mortality and functional impairment among the adult populations globally. Only a minority of ischemic ...

Jianghan University

Understanding the process of relapse to abused drugs and ultimately developing treatments that can reduce the incidence of relapse remains the primary goal for the study of substance dependence. Therefore, exploring the metabolite characteristics during the relapse stage is valuable.

Metabolomics analysis of serum in a rat heroin self-administration model undergoing reinforcement based on H-nuclear magnetic resonance spectra.

Drug dependence and withdrawal syndrome induced by abrupt cessation of opioid administration remain a severe obstacle in the clinical treatment of chronic pain and opioid drug addiction. One of the key symptoms during opioid withdrawal is hyperalgesia. The mechanism of opioid withdrawal-induced hyperalgesia remains unclear. P2X2 and P2X3 receptors, members of P2X receptor subunits, act as the integrator of multiple forms of noxious stimuli and play an important role in nociception transduction of chronic neuropathic and inflammatory pain. The process of P2X2 and P2X3 receptor antagonism inhibits inflammatory hyperalgesia, involving the spinal opioid system. However, the role of P2X receptors involved in opioid withdrawal-induced hyperalgesia has seldom been discussed. To explore the role of P2X2 and P2X3 receptors in the opioid-induced hyperalgesia, heroin self-administration rats were adopted, and the thermal and mechanical nociceptive thresholds were evaluated using the paw withdrawal test after abstinence from heroin for 8 days. In addition, the expressions of P2X2 and P2X3 receptors in dorsal root ganglia were analyzed by immunofluorescence. The results showed that after 8 days of abstinence, heroin self-administration rats showed thermal hyperalgesia and mechanical allodynia. Meanwhile, the expressions of the P2X2 and P2X3 receptors in dorsal root ganglia were increased. These results suggest that upregulation of P2X2 and P2X3 receptors might partially play a role in heroin withdrawal-induced hyperalgesia.

Upregulation of P2X2 and P2X3 receptors in rats with hyperalgesia induced by heroin withdrawal.

Accumulating evidence indicates that P2X receptors may serve an important role in pain and nociceptive sensations. However, recent studies of regulation of P2X receptor expression following nerve injury have produced variable or conflicting results. In the present study the alteration of expression of P2X1-6 receptor subunits in retrograde Flurorogold (FG)-labeled L4+L5 dorsal root ganglion (DRG) neurons were evaluated following unilateral chronic constriction injury (CCI) of the rat sciatic nerve using immunohistochemistry combined with a retrograde fluorescence-tracing method. It was demonstrated that there was no significant difference in the proportion of FG-labeled DRG neurons between the sham and CCI groups (P>0.5). The percentages of P2X1-immunoreactive (IR) and P2X2-IR FG-labeled DRG neurons were not significantly different between the sham and CCI groups (41.5±8.2 vs. 45.2±7.4% and 58.1±6.2 vs. 69.1±3.5%, P>0.05). The percentages of P2X3-IR and P2X6-IR FG-labeled DRG neurons significantly increased in the CCI group compared with the sham group (51.6±4.1 vs. 28.5±3.4% and 41.8±2.2 vs. 22.6±3.3%, P>0.01). By contrast, the percentage of P2X4-IR FG-labeled DRG neurons significantly decreased in the CCI group compared with the sham group (29.4±3.3 vs. 45.0±3.7%, P<0.01). The P2X5-IR positive FG-labeled neurons were not detected in the CCI and sham groups. The results of the present study provided the first evidence regarding the regulation of the expression of the P2X1-6 receptor in sensory neurons being directly associated with chronic nerve injury in rats and also suggest that compared with the P2X3 receptor, the P2X2/3 heteromeric receptor is not the major receptor involved in peripheral neuropathic pain sensation. In addition, the possible functional role of P2X6 receptors in peripheral neuropathic pain requires further investigation.

Alteration of P2X1-6 receptor expression in retrograde Fluorogold-labeled DRG neurons from rat chronic neuropathic pain model.

Electroacupuncture (EA) has effective analgesic effects. Our previous study demonstrated that the upregulation of P2X3 receptors in the dorsal root ganglia (DRG) might participate in heroin withdrawal-induced hyperalgesia. The aim of this study is to further explore whether 2 Hz EA reduces heroin relapse associated with its analgesic effect and whether P2X3 receptors in the DRG are involved in this process. 2 Hz EA was adopted to treat the heroin SA rats in the present study. Heroin-seeking and pain sensitivity were evaluated. The expression of P2X3 receptors in the DRG was detected. Our results showed that compared with the control group, the reinstatement, thermal hyperalgesia, and mechanical allodynia of the heroin-addicted group were increased significantly. The expression of P2X3 receptors in the DRG was increased markedly. After being treated using 2 Hz EA, reinstatement was reduced, hyperalgesia was decreased, and the upregulated expression of P2X3 receptors in the DRG had decreased significantly compared to that in the heroin-addicted group. Consequently, our results indicated that 2 Hz EA was an effective method for treating heroin-induced hyperalgesia and helping prevent relapse, and the potential mechanism might be related to the downregulation of P2X3 receptor expression in the DRG.

2 Hz EA Reduces Heroin Withdrawal-Induced Hyperalgesia and Heroin Relapse by Downregulating P2X3 Receptors in DRG Neurons.

The distributions of P2X subtypes during peripheral neuropathic pain conditions and their differential roles are not fully understood. To explore these characteristics, the lumbosacral dorsal root ganglion (DRG) in the chronic constriction injury (CCI) sciatic nerve rat model was studied. Retrograde trace labeling combined with immunofluorescence technology was applied to analyze the distribution of neuropathic nociceptive P2X1-6 receptors. Our results suggest that Fluoro-Gold (FG) retrograde trace labeling is an efficient method for studying lumbosacral DRG neurons in the CCI rat model, especially when the DRG neurons are divided into small, medium, and large subgroups. We found that neuropathic nociceptive lumbosacral DRG neurons (i.e., FG-positive cells) were significantly increased in medium DRG neurons, while they declined in the large DRG neurons in the CCI group. P2X3 receptors were markedly upregulated in medium while P2X2 receptors were significantly decreased in small FG-positive DRG neurons. There were no significant changes in other P2X receptors (including P2X1, P2X4, P2X5, and P2X6). We anticipate that P2X receptors modulate nociceptive sensitivity primarily through P2X3 subtypes that are upregulated in medium neuropathic nociceptive DRG neurons and/or via the downregulation of P2X2 cells in neuropathic nociceptive small DRG neurons.

Retrograde Labeling of Different Distribution Features of DRG P2X2 and P2X3 Receptors in a Neuropathic Pain Rat Model.

Current therapies for ischemic stroke are insufficient due to the lack of specific drugs. This study aimed to investigate the protective activity of polyphenol extracts from  against cerebral ischemia-reperfusion induced damage. Polyphenols of ethyl acetate and n-butanol fractions were extracted from . BV2 microglial cells exposed to oxygen-glucose deprivation/reoxygenation and mice subjected to middle cerebral artery occlusion/reperfusion were treated by TPE and TPB. Cell viability, cell morphology, apoptosis, mitochondrial membrane potential, enzyme activity and signaling pathway related to oxidative stress were observed. We found that TPE and TPB showed strong antioxidant activity in vitro. The protective effects of TPE and TPB on cerebral ischemia-reperfusion injury were demonstrated by enhanced antioxidant enzyme activities, elevated level of the nucleus transportation of nuclear factor erythroid 2-related factor 2 and expressions of antioxidant proteins, with a simultaneous reduction in cell apoptosis and reactive oxygen species level. In conclusion, TPE and TPB exert neuroprotective effects by stimulating the Nrf2 signaling pathway, thereby inhibiting apoptosis.

Neuroprotective Effect of Polyphenol Extracts from  Retz. against Cerebral Ischemia-Reperfusion Injury.

Excessive reactive oxygen species (ROS) production contributes to brain ischemia/reperfusion (I/R) injury through many mechanisms including inflammation, apoptosis, and cellular necrosis. Chebulic acid (CA) isolated from  has been found to have various biological effects, such as antioxidants. In this study, we investigated the mechanism of the anti-hypoxic neuroprotective effect of CA in vitro and in vivo. The results showed that CA could protect against oxygen-glucose deprivation/reoxygenation (OGD/R) induced neurotoxicity in SH-SY5Y cells, as evidenced by the enhancement of cell viability and improvement of total superoxide dismutase (T-SOD) in SH-SY5Y cells. CA also attenuated OGD/R-induced elevations of malondialdehyde (MDA) and ROS in SH-SY5Y cells. Nuclear factor-E2-related factor 2 (Nrf2) is one of the key regulators of endogenous antioxidant defense. CA acted as antioxidants indirectly by upregulating antioxidant-responsive-element (ARE) and Nrf2 nuclear translocation to relieve OGD/R-induced oxidative damage. Furthermore, the results showed that CA treatment resulted in a significant decrease in ischemic infarct volume and improved performance in the motor ability of mice 24 h after stroke. This study provides a new niche targeting drug to oppose ischemic stroke and reveals the promising potential of CA for the control of ischemic stroke in humans.

Chebulic Acid Prevents Hypoxia Insult via Nrf2/ARE Pathway in Ischemic Stroke.

To study the effect and mechanism of electroacupuncture at "Baihui, Yintang and Shuigou" acupoints on learning and memory in Post-Stroke Cognitive Impairment (PSCI) mice.

Changlong Leng

Wuhan Institute of Biomedical Sciences,
School of Medicine,
Wuhan Institute of Biomedical Sciences, School of Medicine

Jianghan University

Induction of Acute Ischemic Stroke in Mice Using the Distal Middle Artery Occlusion Technique

Changlong Leng

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Wuhan Institute of Biomedical Sciences,School of Medicine,Wuhan Institute of Biomedical Sciences, School of Medicine

Jianghan University

Induction of Acute Ischemic Stroke in Mice Using the Distal Middle Artery Occlusion Technique

Wuhan Institute of Biomedical Sciences,
School of Medicine,
Wuhan Institute of Biomedical Sciences, School of Medicine