Aerobic exercise combined with huwentoxin-I upregulates phase-Ⅱ detoxification enzymes to alleviate obstructive jaundice-induced central nervous system injury in mice / 南方医科大学学报

OBJECTIVE@#To explore the effects of aerobic exercise combined with huwentoxin-I (HWTX-I)-mediated Keap1-Nrf2-ARE pathway on phase II detoxification enzymes HO-1 and NQO1 and their protective effects against obstructive jaundice (OJ)-induced central nervous system injury in mice.@*METHODS@#50 male KM mice were randomly divided into blank group (GO), model group (M), aerobic exercise group (T), HWTX-I group (H), and aerobic exercise combined with HWTX-I group (TH). Mouse models of OJ were established with surgical suture for 72 h in the mice in all the groups except for the blank control group. The mice received interventions by aerobic exercise and tail vein injection of HWTX-I (0.05 μg/g) and were assessed by behavioral observation, Clark's neurological function scores, enzyme-linked immunosorbent assay (ELISA), brain tissue Nissl staining, hippocampal tissue Western blotting, and liver tissue mRNA expression profiling and sequencing.@*RESULTS@#The mice in group M had obvious jaundice symptoms after the operation with significantly increased Clark's neurological score ( < 0.01). Compared with those in group M, the mice in group T, group H, and group TH showed significantly decreased serum levels of ALT, AST, TBIL, and TBA ( < 0.01) with increased contents of 5-HT and BDNF and decreased contents of S100B and NSE in the hippocampus ( < 0.01). Synergistic effects between aerobic exercise and HWTX-I were noted on the above parameters except for the liver function indicators. Interventions with aerobic exercise and HWTX-I, alone or in combination, obviously lessened pathologies in the brain tissue induced by OJ, and the combined treatment produced the strongest effect. The treatment also increased the expression levels of Nrf2, HO-1, and NQO1 mRNA and protein in brain tissues ( < 0.01 or 0.05) with a synergistic effect between aerobic exercise and HWTX-I. Illumina high-throughput sequencing showed that the differentially expressed factors participated mainly in such neural regulatory pathways as neuroactive ligand-receptor interaction, GABAergic synapses, dopaminergic synapses, synaptic vesicle circulation, and axon guidance, involving tissue cell neuronal signal transduction, apoptosis inhibition, immune response, and toxicity. Aerobic exercise and HWTX-I synergistically increased the accumulation of the signal pathways related with neuron damage repair and proliferation.@*CONCLUSIONS@#Aerobic exercise combined with HWTX-I can up-regulate the expression of phase Ⅱ detoxification enzymes HO-1 and NQO1 through the Keap1-Nrf2-ARE pathway to protect the central nervous system against OJ-induced damage in mice.

The diagnostic value of lupus anticoagulant ratio, D-dimer and soluble endothelial protein C receptor for early deep vein thrombosis / 中国医师进修杂志

Objective:To investigate the early predictive value of lupus anticoagulant (LA) ratio, D-dimer (D-D) and soluble endothelial protein C receptor (sEPCR) on deep vein thrombosis (DVT).Methods:Thirty hundred and fifty patients who performed surgery for lower extremity fracture and suspected DVT in Zhejiang Rongjun Hospital from October 2018 to October 2019 were enrolled. With deep vein contrast of the lower extremity as the gold standard, 82 patients with confirmed DVT were treated as the observation group and 268 patients without DVT as the control group. The levels of LA, D-D and sEPCR of two groups were detected by coagulation, immunoturbidimetry and enzyme linked immunosorbent assay—sandwich technique respectively. Indexes of the two groups were compared. Pearson correlation was used to analyze the relationship between plasma levels of LA, D-D and sEPCR, and the predictive value of plasma sEPCR, LA ratio and D-D level on DVT was evaluated by receiver operator characteristic (ROC) curve.Results:The plasma sEPCR, LA ratio and D-D levels in the observation group were significantly higher than those in the control group [(143.30 ± 11.28) μg/L vs.(112.56 ± 14.62) μg/L, 1.51 ± 0.24 vs. 1.22 ± 0.18, (1 013.00 ± 319.54) μg/L vs. (425.17 ± 100.36) μg/L] with statistically significant differences ( P < 0.05). There was no significant differences in activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT) between the two groups ( P > 0.05). In the observation group, plasma sEPCR level was positively correlated with LA ratio and D-D level ( r = 0.280, P = 0.011; r = 0.563, P < 0.001), and LA ratio was positively correlated with D-D level( r = 0.741, P < 0.001). The area under curve (AUC) of D-D in diagnosis of DVT was 0.940, and the sensitivity and specificity were 87.80% and 87.69% when the cut-off value was 569.43 μg/L. The AUC of LA ratio in the diagnosis of DVT was the smallest, which was 0.912, the sensitivity and specificity were 87.80% and 91.25% when the cut-off value was 1.23. The sensitivity was 95.12% and specificity was 95.00% of sEPCR and LA ratio combined with DD in diagnosis of DVT. Conclusions:LA and D-D combined with sEPCR has high predictive value for DVT.

Effects of huwentoxin on tumor necrosis factor apoptotic pathway in the hippocampus of a rat model of cerebral ischemia / 中国组织工程研究

BACKGROUND:Ion channel analytical technique has verified that huwentoxin is an N-type Ca2+channel blocker affecting on presynaptic membrane. OBJECTIVE:To observe the effects of N-type Ca2+channel blocker huwentoxin on expressions of tumor necrosis factorα, tumor necrosis factor receptor I, tumor necrosis factor receptor-related death domain, Fas-related death domain protein and Caspase 8 in the hippocampi of rat models of global cerebral ischemia reperfusion injury. METHODS:Rat models of global cerebral ischemia and subarachnoid catheter were established using Pulsinel i 4-vessel occlusion and then received infusion of huwentoxin or normal saline via a PE10 tube. Morphological changes in the mitochondria and ultrastructure of pyramidal neurons in the hippocampal CA1 region of rats with global cerebral ischemia reperfusion injury were observed using electron microscope. The expressions of tumor necrosis factorα, tumor necrosis factor receptor I, tumor necrosis factor receptor-related death domain, Fas-related death domain protein and Caspase 8 were measured using RT-PCR. RESULTS AND CONCLUSION:Huwentoxin could maintain the basic morphology of mitochondria of rats with global cerebral ischemia reperfusion injury and decrease the expressions of tumor necrosis factorα, tumor necrosis factor receptor I, tumor necrosis factor receptor-related death domain, Fas-related death domain protein and Caspase 8 mRNA. Results suggested that huwentoxin as a novel N-type Ca2+channel blocker could block extracellular Ca2+influx, reduce intracellular Ca2+concentration, diminish a series of pathological lesion induced by intracellular Ca2+overload, protect nerve cells, and lessen the injury to nerve cells of hippocampus after ischemia and hypoxia.

Establishment of global cerebral ischemia combined with intrathecal catheterization models in Sprague-Dawley rats / 中国组织工程研究

BACKGROUND: Cerebral ischemia-reperfusion injury can result in irreversible neuronal function loss, whereas intrathecal administration of analgesia and neuroprotective drugs has been frequently used in the clinic. The animal models undergoing intrathecal administration of neuroprotective substances after cerebral injury are the basis of studies on the effects of neuroprotective substances.OBJECTIVE: To establish animal models of global cerebral ischemia combined with intrathecal catheterization for drug admistration. METHODS: Global cerebral ischemia was induced by four-vessel occlusion method and intrathecal catheterization was performed. Rats were randomly assigned to three groups with 10 rats per group: sham-surgery, model, and huwena toxin-Ⅰ (HWTX-Ⅰ). Rat models of global cerebral ischemia were established and intrathecal catheterization for drug administration was performed in the model and HWTX-Ⅰ groups. After model establishment, rats from the HWTX-Ⅰ group received HWTX-Ⅰ(1.0 μL/kg), and rats from the model group received the same amount of physiological saline. At 4 days after ischemia/reperfusion, Nissl staining was performed to observe the morphological changes of pyramidal neurons in rat hippocampal CA1 region. RESULTS AND CONCLUSION: In the sham-surgery group, numerous pyramidal neurons were densely and orderly arranged, endochylema was blue-stained, and Nissl body staining was even. In the HWTX-Ⅰ group, pyramidal neurons were orderly arranged, sparsely distributed, and some neuronal bodies were atrophic and darkly stained. In the model group, pyramidal neurons were disorderly arranged, and sparsely distributed in the whole CA1 region; in addition, a large number of neurons were atrophic and darkly stained. There was a larger degree of morphological change of hippocampal CA1 region pyramidal neurons in the HWTX-Ⅰ group than in the model group. Results indicate that rat models of global cerebral ischemia combined with intrathecal catheterization were successfully established.

Huwentoxin-Ⅰ: Antinociceptive Effects and Its Comparison with ω-Conotoxin-MVIIA on Acute Visceral Pain in Rats / 中国生物化学与分子生物学报

The antinociceptive effect of epidural administration of huwentoxin-I was elucidated in a tonic visceral pain rat model produced by acute colon inflammation. The nociceptive behaviors were induced by perendoscopically injecting dilute formalin (50 μl) into the depth of the colonic wall in rats. Both ω-conotoxinMVIIA and morphine hydrochloride were given epidurally as positive control while saline as negative control.Similar to ω-conotoxin-MVIIA and hydrochloride morphine, the epidural administration of HWTX-Ⅰ significantly reduced the nociceptive responses in a dose-dependent manner in tonic visceral pain rat model ( P ＜ 0.05). The suppression effects of both huwentoxin- Ⅰ and ω-conotoxin-MVIIA at 20 μg/kg were kept steady compared with the saline group and reached their maximum effects at the doses of 50 ～ 75 μg/kg within 1 hour when the nociception had been observed. It was also found that at the same doses, huwentoxin- Ⅰ was less effective in antinociception than ω-conotoxin-MVIIA. However, ω-conotoxin-MVIIA, but not huwentoxinⅠ , caused an obvious motor dysfunction at these doses. The action of morphine hydrochloride was initiated faster, but lasted for a shorter time than that of huwentoxin- Ⅰ and ω-conotoxin-MVIIA. Thus, huwentoxinⅠ , a potent blocker of neuronal N-type voltage-sensitive calcium channels, induced a remarkable dosedependent restrain effect similar to ω-conotoxin-MVIIA and morphine on the tonic visceral pain produced by colonic wall injection of formalin in conscious rats.