Penehyclidine Hydrochloride Group Research Articles

Penehyclidine hydrochloride alleviates lung ischemia-reperfusion injury by inhibiting pyroptosis

ObjectiveThe aim of this research was to examine how penehyclidine hydrochloride (PHC) impacts the occurrence of pyroptosis in lung tissue cells within a rat model of lung ischemia-reperfusion injury.MethodsTwenty-four Sprague Dawley (SD) rats, weighing 250 g to 270 g, were randomly distributed into three distinct groups as outlined below: a sham operation group (S group), a control group (C group), and a test group (PHC group). Rats in the PHC group received a preliminary intravenous injection of PHC at a dose of 3 mg/kg. At the conclusion of the experiment, lung tissue and blood samples were collected and properly stored for subsequent analysis. The levels of malondialdehyde, superoxide dismutase, and myeloperoxidase in the lung tissue, as well as IL-18 and IL-1β in the blood serum, were assessed using an Elisa kit. Pyroptosis-related proteins, including Caspase1 p20, GSDMD-N, and NLRP3, were detected through the western blot method. Additionally, the dry-to-wet ratio (D/W) of the lung tissue and the findings from the blood gas analysis were also documented.ResultsIn contrast to the control group, the PHC group showed enhancements in oxygenation metrics, reductions in oxidative stress and inflammatory reactions, and a decrease in lung injury. Additionally, the PHC group exhibited lowered levels of pyroptosis-associated proteins, including the N-terminal segment of gasdermin D (GSDMD-N), caspase-1p20, and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3).ConclusionPre-administration of PHC has the potential to mitigate lung ischemia-reperfusion injuries by suppressing the pyroptosis of lung tissue cells, diminishing inflammatory reactions, and enhancing lung function. The primary mechanism behind anti-pyroptotic effect of PHC appears to involve the inhibition of oxidative stress.

Effect of penehyclidine hydrochloride on inflammatory response and oxidative stress in rats with cardiopulmonary bypass related-lung injury.

ABSTRACTPurpose:To investigate the protective effect of penehyclidine hydrochloride (PHC) on cardiopulmonary bypass (CPB)-related lung injury in rats.Methods:Thirty-six rats were divided into control, CPB and PHC groups. The CPB model was established in CPB and PHC groups. In PHC group, 2-mg/kg PHC was added to the pre-filling solution for CPB modeling. At 30 min before CPB (T1), immediately after left hilar opening (T2) and end of experiment (T3), the hemodynamic indexes, blood gas indexes, serum inflammatory factors, lung wet-day ratio and water content and lung tissue oxidative stress indexes were determined.Results:At T2 and T3, compared with CPB group, in PHC group the heart rate and mean arterial pressure increased significantly, the oxygenation index increased significantly, the respiratory index decreased significantly, and the lung wet-day ratio and water content decreased significantly. At T3, compared with CPB group, in PHC groups the serum tumor necrosis factor α, interleukin 6 and interleukin 1β levels decreased significantly, the lung tissue superoxide dismutase level increased significantly, and the myeloperoxidase and malondialdehyde levels decreased significantly.Conclusions:PHC treatment can alleviate the CPB-related lung injury in rats. The mechanisms may be related to its reducing inflammatory response and resisting oxidative stress.

Tumor necrosis factor α‑induced protein 8‑like 2 contributes to penehyclidine hydrochloride pretreatment against lipopolysaccharide‑induced acute lung injury in a mouse model.

The aim of the present study was to investigate the effect of penehyclidine hydrochloride (PHC) pretreatment on mice with lipopolysaccharide (LPS)‑induced acute lung injury (ALI) and its possible underlying mechanisms. Mice were randomly separated into six groups: i)Sham group; ii)LPS group; iii)LPS+PHC group; iv)tumor necrosis factor a‑induced protein 8‑like protein 2 (TIPE2) group; v) LPS+TIPE2 group; and vi) LPS+TIPE2+PHC group. The ALI model was induced using LPS through intratracheal injection. The mice received adenovirus gene to induce the overexpression of TIPE2. After mice were sacrificed, lung injury indices were assessed, and arterial blood, bronchoalveolar lavage fluid and lung tissues were collected for subsequent assays. Expression levels of related proteins were detected by using western blotting. It was found that compared with the sham group, the mice treated with LPS showed increased lung injury and dysfunctions of gas exchange. However, these trends were significantly ameliorated in the LPS+PHC group. Evaluation of protein expression in lung tissues showed that the increased expression of nuclear NF‑κB p65 and p‑c‑Jun N‑terminal kinase (JNK) induced by LPS were suppressed in the LPS+PHC group and the expression of TIPE2 was increased. The mice that received adenovirus gene to induce TIPE2 overexpression could also showed protective effects compared with the mice in the LPS group. However, the expression of TIPE2 decreased rather than increased in LPS group. In the mice pretreated with PHC, the expression of TIPE2 increased in mice with LPS‑induced ALI. To conclude, PHC pretreatment could inhibit the occurrence of inflammation and apoptosis in LPS‑induced ALI. This process may be related to the activation of TIPE2 and the inhibition of NF‑κB and JNK signaling pathway in the lungs of mice.

The Mechanism of Penehyclidine Hydrochloride and Its Effect on the Inflammatory Response of Lung Tissue in Rats with Chronic Obstructive Pulmonary Disease During Mechanical Ventilation.

BackgroundPenehyclidine hydrochloride is a selective antagonist of M1 and M3 receptors. Clinical studies suggest that it is a potential drug for the treatment of chronic obstructive pulmonary disease (COPD). The purpose of this study was to evaluate the effect of penehyclidine hydrochloride on the inflammatory response of lung tissue during mechanical ventilation in rats with COPD and explore the role of the c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) signaling pathway.MethodsEight-week-old male Sprague Dawley rats were exposed to cigarette smoke for 30 minutes every day for two months, and on the first and thirtieth days, 200 ug of lipopolysaccharide was injected into the trachea. Two months later, the rats were randomly divided into the control group (C), model group (M), model + normal saline group (N), and penehyclidine hydrochloride group (H) to undergo anesthesia and mechanical ventilation. In group H, 1 mg/kg of penehyclidine hydrochloride was injected intravenously.ResultsThe results showed that: ① Compared with group C, the other groups all showed typical chronic obstructive pathological changes in the lung tissue; their wet/dry weight ratio (W/D), TNF-α, JNK, and p-JNK levels increased (P < 0.05), and their interleukin (IL)-10 levels decreased (P < 0.05). ② Compared with group M, there was no significant change in the lung tissue indexes in group N (P > 0.05). ③ Compared with group N, the W/D, TNF-α, JNK, and p-JNK levels in group H decreased (P < 0.05), while the levels of IL-10 increased (P < 0.05).ConclusionPenehyclidine hydrochloride can alleviate the pulmonary inflammatory response in rats with COPD undergoing mechanical ventilation. The JNK/SAPK signaling pathway may be involved in this process.

Effect of penehyclidine hydrochloride on IL-6, TNF-α, HIF- 1α and oxidative stress in lung tissue of young rats with acute lung injury

Purpose: To investigate the effect of penehyclidine hydrochloride (PHC) on interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), hypoxia inducible factor-1α (HIF-1α), and oxidative stress levels in lung tissues of acute lung injury (ALI) neonatal rats.Methods: 40 male Sprague-Dawley (SD) rats were assigned to model, low-dose PHC, high-dose PHC, and control groups (n = 10). Levels of IL-6, TNF-α and HIF-1α were evaluated by enzyme-linked immunosorbent assay (ELISA). Pulmonary lesions were determined histologically using H&amp;E staining.Results: The lung tissue levels of IL-6, TNF-α and HIF-1α were significantly higher in model rats than in control rats, and significantly lower in PHC-treated rats than in model group, with decrease in levels as PHC dose increased (p &lt; 0.05). The lung tissue activity of MPO and level of MDA in model rats were significantly higher than those in control rats, but significantly lower in the lung tissues of the two PHC groups than in the model group; decrease in levels occurred as PHC dose increased (p &lt; 0.05).Conclusion: PHC decreases the lung and serum levels of IL-6, TNF-α and HIF-1α in a rat model of ALI, and mitigates pulmonary oxidative stress and lung tissue damage. Thus, penehyclidine hydrochloride may be useful to mitigate ALI-induced damage in patients. However, further studies and clinical trials are required to ascertain this&#x0D; Keywords: Penehyclidine hydrochloride, Alveolar septum, Acute lung injury, Inflammatory cells, IL-6, TNF-α, HIF-1α, Oxidative stress

Analysis of Respiratory Mechanisms in Chronic Obstructive Pulmonary Disease Patients Under the Effects of Penehyclidine Hydrochloride Combined with Continuous Positive Airway Pressure Ventilation

Objective: Our aim was to evaluate the respiratory mechanical effects of penehyclidine hydrochloride combined with intraoperative pulmonary continuous positive airway pressure (CPAP) ventilation on pulmonary function and postoperative pulmonary complications in patients with chronic obstructive pulmonary disease (COPD) during one-lung ventilation. Methods: Eighty patients with COPD who underwent thoracoscopic surgery under general anesthesia were selected and their conditions were validated using chest X-rays. They were then divided into 4 groups in the light of the random number table (n = 20), the regular group (group-C), the penehyclidine hydrochloride group (Group-P), CPAP ventilation group (Group A) and combination group (PA group). All patients underwent mechanical ventilation after double lumen bronchial intubation, inhaled oxygen concentration 100%, oxygen flow 2 L/min, VT 6~8 ml/kg, RR 10~14 times/min, single lung ventilation (OLV) RR 12~16 times/min, maintaining PETCO2 40~45 mmHg. Before the endotracheal intubation, the P group and the PA group were intravenously infused with penehyclidine hydrochloride 0.01 mg/kg. After the start of the incision, each group was changed to OLV. During the OLV of groups A and PA, the lungs of the operation side were CPAP. The pressure is 2 cm H2O. The respiratory mechanics parameters (Ppeak, Pplat , Raw, Cdyn) were monitored immediately before the incision (T1), OLV 60 min (T2), and OLV 120 min (T3). The blood gas analysis of the radial artery was collected at T2, and the OI, RI was calculated. VD/VT , Aa DO2; venous blood was collected at 120 minutes (OL) of OLV, serum concentrations IL-10, TNF-α, IL-6 were detected by EELISA; pulmonary complications occurred within 72 hours after operation. Results: Compared with group C, the serum concentrations TNF-α, IL-6, IL-10, Ppeak, Pplat , Raw, RI, VD/VT, A-aDO2 and pulmonary complications were lower in group P and PA. Cdyn and OI increased (P &lt; 0.05). The serum concentrations IL-10, TNF-α, IL-6 in group A decreased, the incidence of RI, VD/VT, A-aDO2 and pulmonary complications decreased, and OI increased (P &lt; 0.05); compared with group-P or group-A, the concentration of serum IL-10, TNF-α, IL-6 in the PA group, RI, VD/VT, A-aDO2 and pulmonary complications decreased, and OI increased (P &lt; 0.05). Conclusion: Compared with the single application, penehyclidine hydrochloride combined with CPAP ventilation can improve lung function, inhibit inflammation and reduce lung injury in patients with COPD during thoracoscopic surgery, and help to improve prognosis.

Penehyclidine Hydrochloride Premedication Is Not Associated with Increased Incidence of Post-Operative Cognitive Dysfunction or Delirium:A Systemic Review and Meta-Analysis.

Objective Post-operative cognitive dysfunction (POCD) and post-operative delirium (POD) are two common post-operative cerebral complications. The current meta-analysis was to systematically review the effects of penehyclidine hydrochloride (PHC) on POCD and POD in surgical patients.Methods Electronic databases were searched to identify all randomized controlled trials comparing PHC with atropine/scopolamine/placebo on POCD and POD in surgical patients. Primary outcomes of interest included the incidences of POCD and POD; the secondary outcomes of interest included peri-operative mini-mental state examination (MMSE) scores. Two authors independently extracted peri-operative data, including patients' baseline characteristics, surgical variables, and outcome data. For dichotomous data (POCD and POD occurrence), treatment effects were calculated as odds ratio (OR) and 95% confidential interval (CI). Each outcome was tested for heterogeneity, and randomized-effects or fixed-effects model was used in the presence or absence of significant heterogeneity. For continuous variables (MMSE scores), treatment effects were calculated as weighted mean difference (WMD) and 95% CI. Statistical significance was defined as P<0.05.Results Our search yielded 33 studies including 4017 patients. Meta-analysis showed that, the incidence of POCD in PHC group was comparable to that in saline group (OR=0.97; 95% CI: 0.58-1.64; P=0.92), scopolamine group (OR=0.78; 95% CI: 0.48-1.27; P=0.32) and atropine group (OR=1.20; 95% CI: 0.86-1.67; P=0.29). The incidence of POD in PHC group was comparable to that in saline group (OR=1.53; 95% CI: 0.81-2.90; P=0.19) and scopolamine group (OR=0.53; 95% CI: 0.06-4.56; P=0.56), but higher than that in atropine group (OR=4.49; 95% CI: 1.34-15.01; P=0.01).Conclusions PHC premedication was not associated with increased incidences of POCD or POD as compared to either scopolamine or placebo.

Effect of penehyclidine hydrochloride on hyperoxia-induced acute lung injury and the relationship with endoplasmic reticulum stress-dependent apoptosis in infantile rats

Objective To evaluate the effect of penehyclidine hydrochloride on hyperoxia-induced acute lung injury and the relationship with endoplasmic reticulum stress-dependent apoptosis in infantile rats. Methods Ninety-six clean-grade male infantile Sprague-Dawley rats, weighing 40-50 g, aged 14 days, were allocated into 4 groups (n=24 each) using a random number table method: control group (C group), penehyclidine hydrochloride group (P group), hyperoxia group (HO group) and hyperoxia plus penehyclidine hydrochloride group (HP group). Infantile rats were intravenously injected with penehyclidine hydrochloride (0.3 mg/kg) at the same time point every day for 3 consecutive days in P and HP groups.Infantile rats were injected with the equal volume of normal saline instead of penehyclidine hydrochloride in C and H groups.Acute lung injury was induced by inhaling oxygen at concentration greater than 90% for 72 h starting from 4th day after administration.Infantile rats were sacrificed at the end of inhaling, and lungs were removed for examination of the pathological changes and for determination of weight to dry weight ratio (W/D ratio), index of quantitative evaluation for alveolar damage (IQA), pneumonocyte apoptosis (using TUNEL), expression of glucose-regulated protein 78 (GRP78) and CCAAT/enhancer-binding protein homologous protein(CHOP) protein and mRNA in lung tissues (by Western blot or using reverse transcription-polymerase chain reaction). The mitochondrial injury score was assessed, and apoptotic index (AI) was determined. Results Compared with C group, the W/D ratio, IQA, AI and mitochondrial injury score were significantly increased, and the expression of GRP78 and CHOP protein and mRNA was up-regulated in HO and HP groups(P 0.05). Compared with HO group, the W/D ratio, IQA, AI and mitochondrial injury score were significantly decreased, the expression of GRP78 and CHOP protein and mRNA was down-regulated (P<0.05), and the pathological changes of lung tissues were significantly attenuated in HP group. Conclusion Penehyclidine hydrochloride can mitigate hyperoxia-induced acute lung injury, and the mechanism may be related to inhibiting endoplasmic reticulum stress-dependent apoptosis in infantile rats. Key words: Cholinergic antagonist; Acute lung injury; Hyperoxia; Endoplasmic reticulum stress; Apoptosis

Effects of penehyclidine hydrochloride on Nrf2/ARE signaling pathway during endotoxin-induced acute lung injury in neonate rats

Objective: To evaluate the effects of penehyclidine hydrochloride on the nuclear factor erythroid 2-related factor 2/antioxidant responsive element (Nrf2/ARE) signaling pathway during endotoxin-induced acute lung injury(ALI) in neonate rats. Methods: Forty 7-day-old Wistar rats weighing 12-18 g were randomly divided into 4 groups (n=10) using a random number table: normal saline group(NS group), acute lung injury(ALI group), penehyclidine hydrochloride group(PHC group) and penehyclidine hydrochloride+ Nrf2 siRNA plasmid group(PNS group). The ALI model was induced with intraperitoneal endotoxin (5.0 mg/kg) in groups ALI, PHC and PNS. In groups PHC and PNS, penehyclidine hydrochloride (2.0 mg/kg) was injected intraperitoneally at 1 h before ALI respectively, while the equal volume of normal saline was administered in groups NS and ALI. The animal of PNS group were inhaled adenovirus packaging of Nrf2-siRNA three times (one time a day) before modeling. At 4 h after endotoxin injection, the rats were sacrificed. The lungs were collected to determine the wet/dry(W/D) lung weight ratio. The expression of Nrf2 and heme oxygen and enzyme 1(HO-1) were determined by Western blotting, contents of tumor necrosis factor-alpha(TNF-α),interleukin10 (IL-10)were determined by enzyme-linked immunosorbent assay(ELISA). The cell apoptosis were determined by transferase-mediated deoxyuridine triphosphate-biotin nick end labeling(TUNEL),and the apoptotic index was calculated. Results: The W/D ratio in NS, ALI, PHC and PNS groups were (4.2±0.1), (9.6±0.7), (6.5±0.6), (8.3±1.3) respectively. The apoptotic index were (3.7±0.5)%, (31.5±3.2)%, (17.6±4.2)%, (28.1±3.5)%respectively.The contents of TNF-α were (10.3±1.6), (98.5±8.5), (68.5±6.7), (89.9±8.5) pg/ml respectively. The contents of IL-10 were (7.9±0.6), (102.8±9.3), (72.5±5.8), (97.7±9.1) pg/ml respectively.The expression of Nrf2 were (23.2±7.6), (79.8±13.0), (155.5±16.7), (12.0±3.3) respectively. The expression of HO-1 were (31.7±8.6), (90.8±10.3), (147.6±22.5), (61.4±9.7) respectively. There were statistically significant differences among different groups (F=86.013, 154.897, 328.810, 374.198, 333.965, 125.274, all P<0.05). Compared with group NS, the W/D ratio, apoptotic index and the contents of TNF-α, IL-10 increased, the expression of Nrf2 and HO-1 up-regulated in group ALI and group PHC (all P<0.05). Compared with group ALI, the W/D ratio, apoptotic index and the contents of TNF-α, IL-10 decreased, the expression of Nrf2 and HO-1 up-regulated in group PHC (all P<0.05). Compared with group ALI, no significant differences were found in the W/D ratio, apoptotic index and the contents of TNF-α, IL-10 in group PNS(all P>0.05), while the expression of Nrf2 and HO-1 down-regulated in group PNS (all P<0.05). Compared with group PHC, the W/D ratio,apoptotic index and the contents of TNF-α,IL-10 increased, the expression of Nrf2 and HO-1 down-regulated in group PNS (all P<0.05). Conclusion: Nrf2/ARE signaling pathway is involved in the reduction of ALI by penehyclidine hydrochloride in neonate rats.

Effects of penehyclidine hydrochloride combined with unilateral high-frequency jet ventilation on pulmonary function and inflammatory response during one-lung ventilation in patients with chronic obstructive pulmonary disease

Objective To evaluate the effects of penehyclidine hydrochloride (PHCD) combined with high-frequency jet ventilation (HFJV) of the operated lungs on pulmonary function and inflammatory response during one-lung ventilation (OLV) in patients with chronic obstructive pulmonary disease (COPD). Methods Sixty American Society of Anesthesiologists physical status Ⅱ or Ⅲ patients of both sexes with COPD, aged 40-64 yr, with body mass index of 17-26 kg/m2, scheduled for elective video-assisted thoracoscopic surgery under general anesthesia, were divided into 4 groups (n=15 each) using a random number table method: conventional group (group C), PHCD group (group P), HFJV group (group H) and PHCD combined with HFJV group (group PH). After induction of anesthesia, the patients were intubated with a double-lumen tube and then mechanically ventilated, with inspired oxygen concentration 100%, oxygen flow rate 2 L/min, tidal volume 6-8 ml/kg, respiratory rate 10-14 breaths/min, and respiratory rate was set at 12-16 breaths/min and PETCO2 was maintained at 40-45 mmHg during OLV.PHCD 0.01 mg/kg was intravenously injected before intubation in P and PH groups.The ventilation mode was changed to OLV after beginning of skin incision in each group.Ventilation of the lung on the operated side was performed by means of HFJV(driving pressure 0.5 kg/cm2)during OLV in P and PH groups.The pneumodynamic parameters such as airway peak pressure (Ppeak), airway plateau pressure (Pplat), airway resistance (Raw) and dynamic lung compliance (Cdyn) were monitored immediately before skin incision (T1) and at 60 and 120 min of OLV (T2, 3). Arterial blood samples were obtained at T2 for blood gas analysis, and oxygenation index (OI), respiratory index (RI), physiologic dead space fraction (VD/VT) and alveolar-arterial oxygen gradiant (A-aDO2) were calculated.Venous blood samples were drawn at T3 for determination of the serum concentrations of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-8 by enzyme-linked immunosorbent assay.The occurrence of pulmonary complications was recorded within 72 h after operation. Results Compared with group C, the serum concentrations of TNF-α, IL-6 and IL-8, Ppeak, Pplat, Raw, RI, VD/VT, A-aDO2 and incidence of pulmonary complications were significantly decreased, and Cdyn and OI were increased in group P and group PH (P<0.05), and the serum concentrations of TNF-α, IL-6 and IL-8, RI, VD/VT, A-aDO2 and incidence of pulmonary complications were significantly decreased, and OI was increased in group H (P<0.05). Compared with group P or group H, the serum concentrations of TNF-α, IL-6 and IL-8, RI, VD/VT, A-aDO2 and incidence of pulmonary complications were significantly decreased, and OI was increased in group PH (P<0.05). Conclusion PHCD combined with HFJV of the operated lung produces better efficacy in improving respiratory function, inhibiting inflammatory responses, reducing lung injury and is more helpful in improving prognosis than either alone in the patients with COPD. Key words: Cholinergic antagonists; High-frequency jet ventilation; Pulmonary disease, chronic obstructive; Respiratory function tests; Systemic inflammatory response syndrome

Role of M3 receptors in penehyclidine hydrochloride-induced reduction of endotoxin-induced injury to human pulmonary microvascular endothelial cells

Objective To evaluate the role of M3 receptors in penehyclidine hydrochloride (PHC)-induced reduction of lipopolysaccharide (LPS)-induced injury to human pulmonary microvascular endothelial cells (PMVECs). Methods Human PMVECs transfected with M3 shRNA were seeded in 6-well plates (2 ml/hole) or in culture flasks (4 ml/flask) at the density of 1×105 cells/ml and divided into 5 groups (n=5 each) using a random number table method: control group (group C), LPS group, PHC plus LPS group (group P+ LPS), LPS plus M3 shRNA transfection group (group LPS+ shRNA), and PHC plus LPS plus M3 shRNA transfection group (group P+ LPS+ shRNA). Group C received no mediation, and LPS was added at the final concentration of 0.1 μg/ml in the other groups.PHC 2 μg/ml was added at 1 h before adding LPS in P+ LPS and P+ LPS+ shRNA groups.The cells were transfected with plasmid containing 2.5 nmol/L M3 receptor shRNA in LPS+ shRNA group and P+ LPS+ shRNA group.Contents of filamentous actin (F-actin) in endothelial cells were measured by flow cytometry at 1 h after adding LPS.The expression of myosin light chain kinase (MLCK) and VE-cadherin protein was examined by immunofluorescence.The expression of nuclear factor kappa B (NF-κB) p65 and IκB was detected by Western blot.Contents of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were determined by enzyme-linked immunosorbent assay.The M3 receptor mRNA transcription was detected by real-time polymerase chain reaction at 10, 30 and 60 min after adding LPS. Results Compared with group C, F-actin content was significantly decreased, the expression of VE-cadherin and IκB was down-regulated, the contents of TNF-α and IL-6 were increased, and the expression of MLCK and NF-κB p65 was up-regulated in LPS and P+ LPS groups (P 0.05). Compared with group LPS, F-actin content was significantly increased, the expression of VE-cadherin and IκB was up-regulated, the contents of TNF-α and IL-6 were decreased, and the expression of MLCK, NF-κB p65 and M3 receptor mRNA was down-regulated in group P+ LPS and group LPS+ shRNA (P<0.05). Compared with group P+ LPS, F-actin content was significantly increased, the expression of VE-cadherin and IκB protein was up-regulated, TNF-α and IL-6 contents were decreased, and the expression of MLCK, NF-κB p65 and M3 receptor mRNA was down-regulated in group P+ LPS+ shRNA (P<0.05). Conclusion PHC reduces LPS-induced injury to human PMVECs through interfering with M3 receptors and inhibiting NF-κB-mediated inflammatory responses. Key words: Receptor, muscarinic M3; Cholinergic antagonists; Endotoxins; Endothelial cells

Effect of penehyclidine hydrochloride injection on pulmonary ischemia-reperfusion in aortic dissection surgery

Objective: To evaluate the effect of Penehyclidine Hydrochloride Injection on pulmonary ischemia-reperfusion in aortic dissection surgery. Methods: This study was a prospective randomized controlled trial. Between September 2015 and October 2017 in the Second Affiliated Hospital of Zhengzhou University, 60 patients with aortic dissection underwent total arch replacement surgery were randomly divided into penehyclidine hydrochloride group (group A 30 cases) and control group (group B 30 cases). Patients in group A were infused with penehyclidine hydrochloride 0.05 mg/kg after entering the operating room. Patients in group B were infused with the same dose of 0.9% normal saline. Serum tumor necrosis factor (TNF-α), interleukin-6 (IL-6), interleukin-1 (IL-1) and oxygenation index (OI) before anesthesia (T1), before cardiopulmonary bypass (T2), 1 h after cardiopulmonary bypass (T3), the end of cardiopulmonary bypass (T4), 4 h after surgery (T5) and 24 h after surgery (T6) were measured. The time of postoperative ventilator support and ICU staywas recorded. Results: At the time of T4, T5 and T6, the serum levels of TNF-α[(0.10±0.08), (0.13±0.02), (0.23±0.17) mg/L], IL-6[(0.23±0.08), (0.34±0.07), (0.54±0.17) mg/L]and IL-1[(0.62±0.14), (1.02±0.27), (1.44±0.40) mg/L]in group A were significantly lower than those in group B[(0.30±0.09), (0.51±0.19), (0.86±0.02) mg/L; (0.73±0.19), (1.33±0.13), (1.98±0.13) mg/L; (0.93±0.19), (1.43±0.66), (2.04±0.45) mg/L]. The OI was higher in group A[(446.7±267.0), (386.7±169.5), (391.7±227.9) mmHg]than that of group B[(341.2±145.2), (299.5±98.7), (275.0±127.3) mmHg](P<0.05). The time of ventilator support and ICU stay after operation was (3.6 ±1.2)d, (8.4 ± 2.0) d in group A, which was shorter than that in group B[(4.3 ± 1.8), (10.0 ± 2.2) d], and there were statistical differences between the two groups (all P<0.05). Conclusions: In total arch replacement, intravenous penehyclidine hydrochloride injection may decrease the release of serum TNF-α, IL-6, IL-1, improve oxygenation index, reduce lung ischemia-reperfusion injury, shorten the time of ventilator support and ICU stay after operation, and thus improve the prognosis of patients.

Effect of penehyclidine hydrochloride pretreatment on Nrf2/HO-1 signaling pathway in renal tissues of rats with rhabdomyolysis-induced acute kidney injury

Objective To investigate the effect of penehyclidine hydrochloride (PHC) pretreatment on nuclear factor erythroid 2-related factor 2/heme oxygenase-1 (Nrf2/HO-1) signaling pathway in renal tissues of rats with rhabdomyolysis-induced acute kidney injury (AKI). Methods Thirty-six pathogen-free male Sprague-Dawley rats, weighing 200-220 g, were assigned into 3 groups (n=12 each) using a random number table: control group (group C), group AKI and PHC pretreatment group(group PHC). Rhabdomyolysis was induced by intramuscular injection of 50% glycerol 10 ml/kg in bilateral hindlimbs.PHC 0.2 mg/kg was injected intraperitoneally at 30 min before glycerol was injected intramuscularly in group PHC.At 1 and 6 h after glycerol injection, serum was collected for determination of blood urea nitrogen(BUN) and creatinine(Cr) concentrations, and bilateral kidneys were harvested for pathological examination and for determination of HO-1 activity and expression of Nrf2 mRNA and HO-1 mRNA (by quantitative real-time polymerase chain reaction), Nrf2 in nucleoprotein and total protein and HO-1 in total protein in renal tissues (by Western blot). The damage to the renal tubules was scored. Results Compared with group C, the BUN and Cr concentrations in serum and renal tubular damage scores were significantly increased, the expression of Nrf2 in nucleoprotein and total protein and HO-1 in total protein was significantly up-regulated, and HO-1 activity was significantly increased in AKI and PHC groups, the expression of HO-1 mRNA was significantly up-regulated in group AKI, and the expression of Nrf2 mRNA and HO-1 mRNA was significantly up-regulated in group PHC (P<0.01 or 0.05). Compared with group AKI, the BUN and Cr concentrations in serum and renal tubular damage scores were significantly decreased, the expression of Nrf2 in nucleoprotein and total protein and HO-1 in total protein was significantly up-regulated, and HO-1 activity was significantly increased in group PHC (P<0.01 or 0.05). Conclusion The mechanism by which PHC pretreatment attenuates rhabdomyolysis-induced AKI may be related to activation of Nrf2/HO-1 signaling pathway in renal tissues of rats. Key words: Penehyclidine hydrochloride; Rhabdomyolysis; Kidney; Nuclear factor erythroid 2-related factor 2; Heme oxygenase-1

Role of PI3K/Akt signaling pathway in penehyclidine hydrochloride-induced inhibition of autophagy in intestinal tissues in a rat model of limb ischemia-reperfusion

Objective To investigate the role of phosphatidylinositol 3-kinase/serine-threonine kinase (PI3K/Akt) signaling pathway in penehyclidine hydrochloride-induced inhibition of autophagy in intestinal tissues in a rat model of limb ischemia-reperfusion. Methods Sixty-four healthy adult male SPF Sprague-Dawley rats, weighing 220-250 g, were randomly divided into 4 groups (n=16 each) using a random number table: sham operation group (group S), limb ischemia-reperfusion group (group LIR), penehyclidine hydrochloride group (group PHC), and penehyclidine hydrochloride + PI3K inhibitor 3-methyladenine group (group PM). The animals were anesthetized with sevoflurane, and bilateral femoral arteries were clamped for 3 h followed by 4 h of reperfusion in LIR, PHC and PM groups.At 15 min before reperfusion, normal saline 1 ml was injected intravenously in S and LIR groups, penehyclidine hydrochloride 0.15 mg/kg was injected intravenously in group PHC, and penehyclidine hydrochloride 0.15 mg/kg was injected intravenously, and 3-methyladenine 10 mg/kg was injected intraperitoneally at the same time in group PM.At 4 h of reperfusion, venous blood samples were collected for determination of serum lipopolysaccharide (LPS) and lactic dehydrogenase (LDH) levels.The animals were sacrificed after blood sampling, and the small intestine was removed for determination of the expression of Beclin-1, microtubule-associated protein 1 light chain 3 Ⅰ (LC3Ⅰ) and LC3Ⅱ in intestinal tissues by Western blot.The ratio of LC3Ⅱ expression to LC3Ⅰexpression (LC3Ⅱ/LC3Ⅰ) was calculated. Results Compared with group S, the serum LPS and LDH levels were significantly increased, the expression of Beclin-1 in intestinal tissues was up-regulated, LC3Ⅱ/LC3Ⅰ was increased (P 0.05). Compared with group LIR, the serum LPS and LDH levels were significantly decreased, the expression of Beclin-1 in intestinal tissues was down-regulated, LC3Ⅱ/LC3Ⅰ was decreased (P 0.05). Compared with group PHC, the serum LPS and LDH levels were significantly increased, the expression of Beclin-1 in intestinal tissues was up-regulated, LC3Ⅱ/LC3Ⅰ was increased (P<0.05 or 0.01), and autophagosomes were increased in group PM. Conclusion The mechanism by which penehyclidine hydrochloride inhibits autophagy in intestinal tissues is related to activation of PI3K/Akt signaling pathway in a rat model of limb ischemia-reperfusion. Key words: 1-Phosphatidylinositol 3-kinase; Protein-serine-threonine kinases; Cholinergic agonists; Extremities; Reperfusion injury; Intestines; Autophagy

Neuroprotective effects of penehyclidine hydrochloride against cerebral ischemia/reperfusion injury in mice

Various reports have suggested that penehyclidine hydrochloride (PHC), a new cholinergic antagonist, exhibits a variety of biological actions such as anti-tumor and cardioprotective effects. This study aimed to investigate the effects of PHC on cerebral ischemia/reperfusion (I/R) injury and evaluate whether the c-Jun N-terminal kinase (JNK)/p38 mitogen-activated protein kinase (p38MAPK) pathway is involved in the protective effects of PHC. Male C57BL/6 mice were randomly assigned to Sham group, ischemia/reperfusion (I/R) group, I/R+PHC (0.1mg/kg) group, and I/R+PHC (1mg/kg) group. Mice were subjected to 2h of transient middle cerebral artery occlusion, followed by 24h of reperfusion except the mice in the sham group. Neurological deficits, infarct volume, brain water content, blood-brain barrier (BBB) integrity, and neuronal apoptosis were evaluated. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), superoxide production, malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) were measured. The expressions of the key proteins in the JNK/p38MAPK pathway were detected using the Western blot. The results suggested that compared to the I/R group, the PHC-treated group showed improved neurological deficits and BBB integrity, and reduced infarction volume, brain water content, and apoptosis. In addition, PHC significantly suppressed the levels of TNF-α, IL-1β, superoxide production, and MDA, and increased the levels of SOD and GSH-Px. Finally, PHC significantly downregulated the phosphorylation of JNK, p38MAPK, and c-Jun, indicating PHC protects against cerebral I/R injury by downregulating the JNK/p38MAPK signaling pathway.

Effect of penehyclidine hydrochloride pretreatment on rhabdomyolysis-induced acute kidney injury in rats

Objective To evaluate the effect of penehyclidine hydrochloride pretreatment on rhabdomyolysis-induced acute kidney injury (AKI) in rats. Methods Forty-two pathogen-free male Sprague-Dawley rats, weighing 200-220 g, aged 2 months, were randomly divided into 3 groups using a random number table: control group (group C, n=6), AKI group (n=18), and penehyclidine hydrochloride group(group PH, n=18). The model of rhabdomyolysis-induced AKI was established by injecting 50% glycerol 10 ml/kg into the lateral muscle of bilateral hindlimbs in AKI and PH groups.The equal volume of normal saline was given in group C. Penehyclidine hydrochloride 0.2 mg/kg was injected intraperitoneally at 30 min before administration of glycerol in group PH.Six rats were selected at 1 h after administration of normal saline in group C, or at 1, 6 and 24 h after administration of glycerol, blood samples were collected from the inferior vena cava for determination of the serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations by enzymic colorimetric method.The animals were sacrificed, and kidney specimens were obtained for pathologic examination and for determination of the expression of DJ-1 and phosphatase tensin homolog deleted on chromosome 10 (PTEN) encoding protein (by immuno-histochemistry and Western blot). The damage to the renal tubules was scored. Results Compared with group C, the serum BUN and Cr concentrations and renal tubular damage score were significantly increased, the expression of DJ-1 was down-regulated, and the expression of PTEN protein was up-regulated in group AKI (P<0.05 or 0.01). Compared with group AKI, the serum BUN and Cr concentrations and renal tubular damage score were significantly decreased, the expression of DJ-1 was up-regulated, and the expression of PTEN protein was down-regulated in group PH (P<0.05 or 0.01). Conclusion Penehyclidine hydrochloride pretreatment can reduce rhabdomyolysis-induced AKI probably by up-regulating the expression of DJ-1 and down-regulating the expression of PTEN protein in rats. Key words: Cholinergic antagonists; Rhabdomyolysis; Kidney failure, acute

The research on protective effect of penehyclidine hydrochloride in rats with myocardial ischemia/reperfusion injury

To compare the protective effect of different dose of penehyclidine hydrochloride (PHC) in rats with myocardial ischemia/reperfusion (I/R) injury. Forty-eight healthy male Wistar rats were randomly divided into six groups (n = 8 each): sham group, sham + 1.0 mg/kg PHC group (sham + H-PHC group), I/R group, I/R + 0.1 mg/kg PHC preconditioning group (I/R + L-PHC group), I/R + 0.3 mg/kg PHC preconditioning group (I/R + M-PHC group), and I/R + 1.0 mg/kg PHC preconditioning group (/R + H-PHC group). 1/R injury model was reproduced by ligation followed by release of the coronary artery, and PHC in different dosages was given' at 30 minutes before model reproduction. At 3 hours after reperfusion, the left ventricular end-diastolic pressure (LVEDP), left ventricular end-systolic pressure (LVESP), ejection fraction (EF), and fractional shortening (FS) were recorded. The levels of aspertate aminotransferase (AST), MB isoenzyme of creatine kinase (CK-MB), and lactate dehydrogenase (LDH) were determined. The myocardial tissues were harvested for the determination of the area at risk (AAR) and the infarct area (AI), and the percentage of AI/AAR was calculated. The examination of myocardial fiber was performed with electron microscopy. Compared with sham and sham + H-PHC groups, LVEDP was increased in I/R groups, LVESP, EF and FS were decreased, and the levels of AST, CK-MB and LDH, as well as the AI/AAR were increased. Compared with I/R group, in pretreatment groups with different doses of PHC, LVEDP was decreased, LVESP, EF and FS were increased, the levels of AST, CK-MB, LDH, and AI/AAR were also decreased, especially in I/R+M-PHC and I/R+H-PHC groups [LVEDP (mmHg, 1 mmHg = 0.133 kPa): 11.33 ±1.17, 9.85 ± 1.09 vs. 15.82 ± 1.79, LVESP (mmHg): 98.9 ± 10.6, 112.8 ± 10.0 vs. 87.8 ± 9.2, EF: 0.681 ± 0.074, 0.741 ± 0.070 vs. 0.569 ± 0.072, FS: (42.4 ± 4.6)%, (46.0 ± 5.1 )% vs. (36.8 ± 3.9)%, AST (U/L): 386.97 ± 80.65, 298.31 ± 54.88 vs. 603.47 ± 173.66, CK-MB (U/L): 3.12 ± 0.84, 2.88 ± 0.72 vs. 7.14 ± 1.54, LDH (U/L): 1,784.23 ± 488.49, 1,629.37 ± 436.34 vs. 2,489.14 ± 460.80, AI/AAR: 0.284 ± 0.014, 0.223 ± 0.008 vs. 0.377 ± 0.011, all P < 0.05]. There was significant difference in LVEDP, LVESP, and AI/AAR between I/R + M-PHC group and I/R + H-PHC group (all P < 0.05), and no significant difference in other parameters (all P > 0.05). It was showed by electron microscopic examination that after I/R injury, the myocyte mitochondria membranes were broken, mitochondria were markedly swollen, mitochondrial cristae disappeared; however in I/R+M-PHC and I/R+H-PHC groups, mitochondrial swelling was mild, the capsule was more or less intact, mitochondrial cristae were partly visible, the structure was complete, especially in the group I/R+H-PHC, and the mitochondrial structure was close to normal. PHC could protect myocardial from I/R injury. Mid dose of PHC (0.3 mg/kg) and high dose of PHC (1.0 mg/kg) could provide better protective effect than low dose of PHC (0.1 mg/kg), and high dose of PHC is better in effect than the middle dose.

Effects of penehyclidine hydrochloride on activities of NF-κB and AP-1 during actue lung injury induced by blunt chest trauma-hemorrhagic shock and resuscitation in rats

Objective To investigate the effects of penehyclidine hydrochloride on activities of nuclear factor kappa B(NF-κB) and activator protein-1 (AP-1) during actue lung injury induced by blunt chest trauma-hemorrhagic shock and resuscitation (HSR) in rats. Methods Thirty male Sprague-Dawley rats, aged 8 weeks, weighing 250-300 g, were randomly assigned into 3 equal groups (n = 10 each) using a random number table: sham operation group (group S), blunt chest trauma-HSR group (group THSR) and penehyclidine hydrochloride group (group PHCD). The model of actue lung injury induced by blunt chest trauma-HSR was induced by dropping a 300 g weight onto a precordium in anesthetized rats.Blood was withdrawn via the femoral artery 5 min later until MAP was decreased to 35-45 mmHg within 15 min and maintained at this level for 60 min, followed by resuscitation.In PHCD group, PHCD 2 mg/kg was injected intravenously at 60 min after hemorrhagic shock.At 6 h after the model was established, blood samples were obtained for measurement of concentrations of tumor necrosis factor-alpha (TNF-α) in serum.The lungs were then removed for determination of lung water content, myeloperoxidase (MPO) activaty (by colorimetric assay), NF-κB and AP-1 activaties (using electrophoretic mobility shift assay) in lung tissues, and for microscopic examination of pathologic changes (under light microscope). The left lung was lavaged, and lung permeability index (LPI) was calculated. Results Compared with S group, lung water content, LPI, serum TNF-α level and activites of MPO, NF-κB and AP-1 were significantly increased in THSR and PHCD groups.Compared with THSR group, lung water content, LPI, serum TNF-α concentrations and activites of MPO, NF-κB and AP-1 were significantly decreased in PHCD group.The pathological damage to lung tissues was significantly reduced in PHCD group as compared with THSR group. Conclusion PHCD can inhibit activities of NF-κB and AP-1 in lung tissues, thus mitigating acute lung injury induced by blunt chest trauma-HSR in rats. Key words: NF-κappa B; Transcription factor AP-1; Cholinergic antagonists; Thoracic injury; Shock, hemorrhagic; Respiratory distress syndrome, adult

Effect of penehyclidine hydrochloride on endotoxemia-induced intestinal injury in neonatal rats

Objective To investigate the effect of penehyclidine hydrochloride on endotoxemia-induced intestinal injury in neonatal rats. Methods Ninety healthy neonatal Sprague-Dawley rats, aged 7 days, weighing 16-20 g, were randomly divided into 3 groups (n=30 each) using a random number table: control group (group C), lipopolysaccharide (LPS) group, and penehyclidine hydrochloride group (group P). Endotoxemia was induced by intraperitoneal LPS 5 mg/kg in LPS and P groups. In group P, penehyclidine 2 mg/kg was injected intraperitoneally at 30 min before and after LPS injection. The equal volume of normal saline was given in group C. At 2, 6 and 12 h after LPS or normal saline administration, 10 rats in each group were randomly chosen and sacrificed. The ileum was removed to detect wet/dry lung weight ratio (W/D ratio) and contents of tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), hypoxia-inducible factor 1 alpha (HIF-1α) and glutamine (Gln), and for examination of the pathologic changes of the ileum with light microscope. Results Compared with group C, the W/D ratio and contents of TNF-α, IL-6 and HIF-1α were significantly increased, and the content of Gln was decreased at each time point in LPS and P groups. Compared with group LPS, the W/D ratio and contents of TNF-α, IL-6 and HIF-1α were significantly decreased, the content of Gln was increased at each time point, and the pathologic changes of the ileum were mitigated in group P. Conclusion Penehyclidine hydrochloride can reduce endotoxemia-induced intestinal injury in the neonatal rats, and down-regulated expression of HIF-1α, up-regulated expression of Gln and attenuated inflammatory responses may be involved in the mechanism. Key words: Cholinergic antagonists; Endotoxemia; Intestine; Infant, newborn

Protective effect of penehyclidine hydrochloride on lipopolysaccharide-induced acute kidney injury in rat.

We aimed to observe the effect of penehyclidine hydrochloride (PHC) on lipopolysaccharide (LPS)-induced acute kidney injury in rats and expression of tight junction proteins ZO-1 and occludin. Adult male Sprague-Dawley (SD) rats were divided randomly (N = 10) into control group (C), LPS group (LPS), low-dose PHC group (L-PHC), and high-dose PHC group (H-PHC). All rats, except C group, received a vena caudalis injection of 5.0 mg/kg LPS; after 30 min, rats in L-PHC and H-PHC groups received a vena caudalis injection of 0.3 and 0.9 mg/kg PHC. After 24 h, tumor necrosis factor (TNF)-α, interleukin (IL)-1β, serum creatinine (Scr), and blood urea nitrogen (BUN) were detected. Histopathological changes and expression of ZO-1 and occludin were observed in renal tissues. Versus levels of TNF-α (38.5 ± 9.0), IL-1β (46.3 ± 12.7), Scr (37.2 ± 9.3), and BUN (6.5 ± 1.1) in control group, those in LPS group, TNF-α (159.0 ± 21.3), IL-1β (130.8 ± 18.7), Scr (98.5 ± 18.2), and BUN (12.8 ± 1.8), increased obviously (P < 0.05), with significantly structural changes and decreases of ZO-1 and occludin. However, TNF-α (111.3 ± 11.6), IL-1β (78.4 ± 14.3), Scr (51.3 ± 12.5), BUN (8.1 ± 1.2) in H-PHC group, and TNF-α (120.8 ± 14.3), IL-1β (92.5 ± 19.0), Scr (56.7 ± 14.7), BUN (9.7 ± 1.6) in L-PHC group were obviously decreased (P < 0.05). PHC has protective effects on acute kidney injury in sepsis, including abatement of renal tissue inflammation and functional improvement, potentially by upregulating ZO-1 and occludin.