Protective Effects Of Penehyclidine Hydrochloride Research Articles

Penehyclidine hydrochloride protects against lipopolysaccharide-induced acute lung injury by promoting the PI3K/Akt pathway.

Acute lung injury (ALI) attracted attention among physicians because of its high mortality. We aimed to determine whether the phosphatidylinositol-3 kinase (PI3K)/protein kinase B (Akt) pathway is involved in the protective effects of penehyclidine hydrochloride (PHC) against lipopolysaccharide (LPS)-induced ALI. H&E staining was used to observed pathological changes in the lung tissues. ELISA was used to evaluate the concentration of inflammatory mediators in the bronchoalveolar lavage fluid (BALF). White-light microscopy was performed to observe the TUNEL-positive nuclei. The viability of NR8383 alveolar macrophages was determined by using CCK-8. The levels of MPO, MDA, SOD, and GSH-Px were analyzed using ELISA kits. Western blotting was used to evaluate the ERS-associated protein levels and the phosphorylation of PI3K and Akt. PHC administration defended against LPS-induced histopathological deterioration and increased pulmonary edema and lung injury scores, while all of these beneficial effects were inhibited by LY. In addition, PHC administration mitigated oxidative stress as indicated by decreases in lung myeloperoxidase (MPO) and malondialdehyde (MDA) concentrations, and increases in glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) concentrations. It also alleviated LPS-induced inflammation. PHC administration attenuated apoptosis-associated protein levels, improved cell viability, and decreased the number of TdT-mediated dUTP Nick-End Labeling (TUNEL)-positive cells. Furthermore, PHC inhibited ERS-associated protein levels. Meanwhile, the protection of PHC against inflammation, oxidative stress, apoptosis, and ERS was inhibited by LY. Moreover, PHC administration increased PI3K and Akt phosphorylation, indicating that the upregulation of the PI3K/Akt pathway, while this pathway was inhibited by LY. PHC significantly activates the PI3K/Akt pathway to ameliorate the extent of damage to pulmonary tissue, inflammation, oxidative stress, apoptosis, and ERS in LPS-induced ALI.

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.

Penehyclidine hydrochloride protects against anoxia/reoxygenation injury in cardiomyocytes through ATP-sensitive potassium channels, and the Akt/GSK-3β and Akt/mTOR signaling pathways.

Penehyclidine hydrochloride (PHC) can protect against myocardial ischemia/reperfusion (I/R) injury. However, the possible mechanisms of PHC in anoxia/reoxygenation (A/R)-induced injury in H9c2 cells remain unclear. In the present study, H9c2 cells were pretreated with PI3K/Akt inhibitor LY294002, ATP-sensitive K+ (KATP) channel blocker 5-hydroxydecanoate (5-HD), PHC, or KATP channel opener diazoxide (DZ) before subjecting to A/R injury. Cell viability and cell apoptosis were determined by cell counting kit-8 assay and annexin V/PI assay, respectively. Myocardial injury was evaluated by measuring creatine kinase (CK) and lactate dehydrogenase (LDH) activities. Intracellular Ca2+ levels, reactive oxygen species (ROS) generation, mitochondrial membrane potential (ΔΨm ), and mitochondrial permeability transition pore (mPTP) were measured. The levels of cytoplasmic/mitochondrial cytochrome c (Cyt-C), Bax, Bcl-2, cleaved caspase-3, KATP channel subunits (Kir6.2 and SUR2A), and the members of the Akt/GSK-3β and Akt/mTOR signaling pathways were determined by western blotting. We found that PHC preconditioning alleviated A/R-induced cell injury by increasing cell viability, reducing CK and LDH activities, and inhibiting cell apoptosis. In addition, PHC preconditioning ameliorated intracellular Ca2+ overload and ROS production, accompanied by inhibition of both mPTP opening and Cyt-C release into cytoplasm, and maintenance of ΔΨm . Moreover, PHC preconditioning activated mitochondrial KATP channels, and modulated the Akt/GSK-3β and Akt/mTOR signaling pathways. Similar effects were observed upon treatment with DZ. Pretreatment with LY294002 or 5-HD blocked the beneficial effects of PHC. These results suggest that the protective effects of PHC preconditioning on A/R injury may be related to mitochondrial KATP channels, as well as the Akt/GSK-3β and Akt/mTOR signaling pathways.

Postconditioning Protection Against Myocardiocyte Anoxia/Reoxygenation Injury From Penehyclidine Hydrochloride.

Background/AimsTo investigate the postconditioning protective effect of penehyclidine hydrochloride (PHC) against anoxia/reoxygenation (A/R) injury in H9c2 cells along with the involved mechanism and timing effect.MethodsWe divided H9c2 cells into 7 groups: control group, A/R group and PHC+A/R groups at 0 min, 5 mins, 10 mins, 20 mins, 30 mins, respectively (treated with 0.1 μm/L PHC at 0 min, 5 mins, 10 mins, 20 mins, 30 mins after the reoxygenation procedure began). Cell apoptosis, oxidative stress, intracellular Ca2+ concentration, mitochondrial membrane potential and mitochondrial permeability transition pore (MPTP) opening were explored. Bcl-2, Bax, Cyt C, caspase-3 and caspase-9 levels were measured.ResultsA/R significantly increased both cell injury and cell apoptosis. PHC showed postconditioning protective effect by attenuating superoxide production, decreasing Ca2+ overload, restraining MPTP activities, restoring mitochondrial membrane potential, regulating cell apoptosis proteins and modulation of mitochondrial pathway. Earlier administration of PHC offered greater postconditioning protective effect.ConclusionH9c2 cells were protected by PHC from A/R injury regardless of timing of PHC administration (0 min, 5 mins, 10 mins, 20 mins, 30 mins). However, earlier administration of PHC resulted in better PHC postconditioning protection.

Smoking Attenuates Efficacy of Penehyclidine Hydrochloride in Acute Respiratory Distress Syndrome Induced by Lipopolysaccharide in Rats.

BackgroundPenehyclidine hydrochloride is a novel drug for acute respiratory distress syndrome. The aim of the study was to reveal the impact of smoking on the efficacy of the drug in rats with acute respiratory distress syndrome.Material/MethodsA 132 Sprague-Dawley rats were used in this study; 72 rats were used in the smoking models. Penehyclidine hydrochloride (3 mg/kg) was injected to induce acute respiratory distress syndrome. Rats were divided into the smoking group and the non-smoking group; these 2 groups were subdivided according to different treatments. The arterial blood gas analysis (PaO2/FiO2) and extent of pneumonedema (wet-to-dry weight ratio) was analyzed to evaluate disease severity. Expressions of mitogen-activated protein kinases (p-p38MAPK, p38MAPK, p-ERK, ERK, p-JNK, and JNK) in lung tissue were measured using western blot assay.ResultsPenehyclidine hydrochloride improved the pneumonedema (wet-to-dry weight ratio) and hyoxemia (PaO2/FiO2) of the disease in non-smoking group (P<0.001, P<0.001 respectively), but not in smoking group (P=0.244, P=0.424 respectively). The drug inhibited the expressions of phospho-p38MAPK and phospho-ERK in non-smoking group (P<0.001, P<0.001 respectively), but not in smoking group (P=0.350, P=0.507 respectively). In the smoking group, blocking the phospho-p38MAPK or phospho-ERK signal pathway by their inhibitors showed a better therapeutic effect on the pneumonedema and hyoxemia compared with the use of penehyclidine hydrochloride (phospho-p38MAPK: P=0.004, P=0.010 respectively; phospho-ERK: P=0.022, P=0.004 respectively).ConclusionsThe study confirmed the protective effect of penehyclidine hydrochloride in acute respiratory distress syndrome, mainly in the non-smoking group, which might be explained by the fact that phospho-p38MAPK and phospho-ERK signal pathways were difficult to inhibit by the drug in the smoking group.

Penehyclidine hydrochloride protects organs through the regulation of inflammatory mediators

Inflammatory reaction is the most common characteristic reactions of tissue or organ injury. Sepsis and ischemia/reperfusion injury in organs can cause the massive release of inflammatory factors, resulting in systemic inflammatory response syndrome or multiple organ dysfunction syndrome. Regulation of inflammatory factors is an important method to ameliorate organ and tissue injury. In the present study, we investigated the regulation of penehyclidine hydrochloride on inflammatory factors, as well as the safety and effectiveness of its protective effects on tissue and organ injury. Recent studies concerning the regulation of the inflammatory response by penehyclidine hydrochloride were summarized, especially the protective effects of penehyclidine hydrochloride on the lung, and the anti-inflammatory protective effect on other organs such as the heart, brain, kidneys and liver. It is necessary to further explore the mechanisms of penehyclidine hydrochloride in tissue and organ protection. Key words: Penehyclidine hydrochloride; Inflammation mediator; Organ protection

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.

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.

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.

Protective effects of penehyclidine hydrochloride on liver injury induced by cardiopulmonary bypass in Guangxi bama miniature pig

Objective To investigate the effects of penehyclidine hydrochloride on liver injury induced by cardiopulmonary bypass (CPB) in Guangxi bamaminiature pig.Methods Twenty-seven Bama miniature pigs weighted 32-40 kg were chosen,three pigs were used for sampling blood,another twentyfour pigs were used for cardiopulmonary bypass (CPB).Twenty-four pigs were randomly divided into three groups (n=8 each):group Sham,group Mod,and group PHC.Propofol,fentanyl and vecuronium were continued by intravenous injection and inhaled isoflurane interruptly for anesthesic maintain.CPB was performed on mild hypothermic beating heart for 120 min.Pnehyclidine hydrochloride 0.1 mg/kg was injected intravenously before 10 min CPB.Blood samples were collected for aspartate aminotransferase (AST),alanine transaminase (ALT),direct bilirubin (DB) and total bilirubin (TB) before CPB (T0),after CPB 1 h (T1),CPB cease (T2) and operation cease (T3).The content of ATP,MDA and SOD in liver tissue at befor and after CPB were measured and the pathologic change of liver was evaluated by electron microscope.Results The levels of ALT,AST,DB and TB in group Mod and group PHC were significantly higher at T1-T3 than those in group Sham (P ＜ 0.05).ATP,SOD were lower and MDA was higher significantly in group Mod and group PHC after CPB than that in group Sham (P ＜ 0.05).Compared with the group Mod,The levels of ALT,AST,DB,TB and MDA were lower and ATP,SOD were higher significantly respectively in group PHC (P ＜ 0.05).The electron microscopec examination showed that histologic damage was significantly reduced in group PHC compared with group Mod.Conclusion Pnehyclidine hydrochloride can attenuate Iiver injury induced by CPB,antioxygen to remove oxygen free radical in liver may be the mechanism. Key words: Penehyclidine hydrochloride ; Cardiopulmonary bypass ; Liver injury ; Antioxygen

Penehyclidine hydrochloride and organ protection during cardiopulmonary bypass

Background Penehyclidine hydrochloride (PHC),which is derived from hyoscyamine is a new anticholinergic drug,has both antimuscarinic and antinicotinic activities and retains potent central and peripheral anticholinergic activities.Clinical data showed that PHC had good curative effect for the pesticides poison of organic phosphorus and soman poisoning,asthma and chronic obstructive pulmonary disease,septic shock,the diseases of gastrointestinal tract and drug addiction,et al.Additionally,recent studies have suggested that PHC has protective effects of hepatic,renal and lung injury and brain ischemia/reperfusion injury (I/RI).Objective The protective effect of PHC on organs during cardiopulmonary bypass (CPB) is briefly reviewed.Content The pharmacological actions of PHC,and effects on organs during CPB and the mechanism of its action are discussed.Trend Further studies of PHC for prevention of organ injury during CPB in patients with open-heart surgery are needed. Key words: Penehyclidine hydrochloride; Organ protection; Cardiopulmonary bypass

Protective effects of penehyclidine hydrochloride on acute lung injury caused by severe dichlorvos poisoning in swine

Background Organophosphate poisoning is an important health problem in developing countries which causes death mainly by inducing acute lung injury. In this study, we examined the effects of penehyclidine hydrochloride (PHC), a selective M-receptor inhibitor, on dichlorvos-induced acute lung injury in swine. Methods Twenty-two female swines were randomly divided into control (n=5), dichlorvos (n=6), atropine (n=6), and PHC (n=5) groups. Hemodynamic data, extravascular lung water index (EVLWI), and pulmonary vascular permeability index (PVPI) were monitored; blood gas analysis and acetylcholinesterase (AchE) levels were measured. PaO2/FiO2, cardiac index (CI), and pulmonary vascular resistance indices (PVRI) were calculated. At termination of the study, pulmonary tissue was collected for ATPase activity determination and wet to dry weight ratio (W/D) testing 6 hours post-poisoning. TUNEL assay, and Bax, Bcl-2, and caspase-3 expression were applied to pulmonary tissue, and histopathology was observed. Results After poisoning, PHC markedly decreased PVRI, increased CI more effectively than atropine. Anticholinergic treatment reduced W/D, apoptosis index (AI), and mitigated injury to the structure of lung; however, PHC reduced AI and caspase-3 expression and improved Bcl-2/Bax more effectively than atropine. Atropine and PHC improved ATPase activities; a significant difference between groups was observed in Ca2+-ATPase activity, but not Na+-K+-ATPase activity. Conclusions The PHC group showed mild impairment in pathology, less apoptotic cells, and little impact on cardiac function compared with the atropine group in dichlorvos-induced acute lung injury.

Penehyclidine hydrochloride preserves the intestinal barrier function in patients undergoing cardiopulmonary bypass

The study objective was to investigate the protective effect of penehyclidine hydrochloride on intestinal barrier function integrity and its therapeutic potential on endotoxemia and systemic inflammatory response in patients undergoing cardiopulmonary bypass. Forty patients undergoing cardiac valve replacement with cardiopulmonary bypass were enrolled in the study. All patients were randomly divided into the penehyclidine hydrochloride or control group (20 patients in each group). Patients in the penehyclidine hydrochloride group received an intravenous injection of 0.05 mg/kg penehyclidine hydrochloride 10 minutes before cardiopulmonary bypass, and those in the control group were given the same volume of saline. Blood samples for blood glucose, lactic acid, intestinal fatty acid binding protein, D-lactate, serum endotoxin (lipopolysaccharide), interleukin-6, and interleukin-10 measurements were collected during the following time points: immediately after anesthesia induction (T0), 10 minutes after the release of aortic-clamping (T1), immediately after weaning from cardiopulmonary bypass (T2), 2 hours postoperatively (T3), 6 hours postoperatively (T4), and 18 hours postoperatively (T5). Blood glucose, lactic acid, intestinal fatty acid binding protein, D-lactate, lipopolysaccharide, interleukin-6, and interleukin-10 were significantly increased at all postoperative time points. At specific postoperative time points, blood glucose, lactic acid, intestinal fatty acid binding protein, D-lactate, lipopolysaccharide, and interleukin-6 were statistically lower in the penehyclidine hydrochloride group than in the control group. Postoperatively, interleukin-10 did not differ between the penehyclidine hydrochloride and control groups. Penehyclidine hydrochloride preserves intestinal barrier function integrity, attenuates endotoxemia, and inhibits systemic inflammatory response in patients undergoing cardiopulmonary bypass, possibly by improving intestinal microcirculation and depressing stress response.

Protective effects of penehyclidine hydrochloride on lipopolysaccharide-induced human umbilical vein endothelial cells injury and its mechanism

Objective To investigate the protective effects of penehyclidine hvdrochloride (PHC)on lipopolysaccharide ( LPS)-induced human umbilical vein endothelial cells (HUVECs) injury and the underlying mechanism.Methods Cultured HUVECs were divided into five groups:control group,LPS group,LPS/PHC ( 10 μg/L) group,LPS/PHC (25 μg/L) group,LPS/PHC (50 μg/L) group.Cell viability was measured by methyl thiazol tetrazolium (MTT) colorimetric method.The concentrations of lactate dehydrogenase (LDH) and nitric oxide (NO) were measured respectively by using chemical colorimetric method and nitrate reduction method.The expression of inducible nitric oxide synthase (iNOS) mRNA and protein was detected hy using reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting respectively.The activity of nuclear factor-KB (NF-KB) and signal transducer and activators of transcription 3 (STAT3) were measured by enzyme linked immunosorbent assay (ELISA).Results As compared with control group,cell viability [(60.31 ± 8.76 )％ vs 100.00％] was decreased and the concentrations of LDH [(326 ±52) μmol/L vs ( 125 ±25) μmoL/L] and NO [(55.49 ± 10.16) μmol/L vs ( 12.13 ±11.02) μmol/L] were increased in LPS group ( P ＜0.01 ) ; PHC could revere these effects on HUVECs (P ＜0.05).At the same time,PHC reduced the expression of iNOS mRNA and protein,and the activity of NF-KB and STAT3 ( P ＜ 0.05 ).Conclusion PHC can protect the endothelial function by reducing the concentration of NO.Inhibiting the NF-KB and STAT3 signal pathway may be the underlying mechanism. Key words: Penehyclidine hydrochloride; Lipopolysaccharide; Human umbilical vein endothelial cells; Inducible nitric oxide synthase; Nitric oxide

Protective effect of penehyclidine hydrochloride on secondary lung injury after traumatic shock in rats

Objective To observe the interfering effect of different doses of penehyclidine hydrochloride (PHC) on the mRNA expressions of nuclear factor kappa B (NF-κB) and inducible nitric oxide synthase (iNOS) in the lung tissue of rats with traumatic shock so as to investigate the protective role of PHC in secondary long injury following traumatic shock and the underlying mechanism.Methods The traumatic shock model was established.A total of 104 Wistar rats were randomly divided into four groups:control group,shock group,low dose PHC group ( P1 group) and high dose PHC group ( P2 group).At the beginning of resuscitation,the rats in P1 and P2 groups were given transjugular intravenous injection of 2 ml/kg isotonic saline containing 0.15 mg/kg and 0- 45 mg/kg PHC respectively,while the rats in shock and control groups were injected only isometric isotonic saline.The rats in the four groups were killed at 2 h,6 h,12 h and 24 h after resuscitation respectively to detect the mRNA expressions of NF-κB and iNOS by using RT-PCR and determine the lung wet/dry weight (W/D) ratio,lung permeability index (LPI) and lung injury score (LIS).Results The mRNA expressions of NF-κB and iNOS,lung W/D ratio,LPI and LIS at all the time intervals in the shock,P1 and P2 groups were all significantly increased as compared with those in the control group (P＜0.05).Howerver,the P2 group showed significant reduction in aspects of the mRNA expressions of NF- κB and iNOS,lung W/D ratio,LPI and LIS at all time points and P1 group also had significant decrease regarding the mRNA expressions of NF-κB and iNOS,lung W/D ratio at2 h,6 h,and LPI and LIS at 2 h,6 h,12 h,as compared with the shock group.Meanwhile,P2 group showed evident decrease at 6 h concerning the mRNA expressions of NF-κB and iNOS,lung W/D ratio,LPI and LIS as compared with P1 group (P ＜ 0.05 ).Conclusions PHC,especially at a large dosage,can significantly mitigate the long injury secondary to traumatic shock,and the mechanism may be associated with the inhibition of mRNA expressions of NF-κB and iNOS. Key words: Shock, traumatic; Nuclear factor kappa B; Nitric oxide synthase; Acute lung injury; Penehyclidine hydrochloride

The protective effects of penehyclidine hydrochloride on intestinal mucosa injury in rabbits with traumatic shock model

Objective To investigate the protective effects of penehyclidine hydrochloride(PHC) on intestinal mucosa injury in rabbits with traumatic shock. Methods The animal model of traumatic shock was established according to Lamson's method.Thirty rabbits were divided into five groups randomly:control group (Con group),traumatic shock group resuscitation with normal saline (NS group),traumatic shock group resuscitation with pcnehyclidine hydrochloride in low dosages (0.05 mg/kg,PHCL group),middle dosages group (0.15 mg/kg,PHCM group) and high dosages group (0.45 mg/kg,PHCH group).At pre-shock,postshock,four time points (T3-T(o)) after resuscitation,mean arterial blood pressure(MAP) and heart rate(HR) were monitored.Plasma diamine oxidase (DAO) activity and lactic acid concentration were measured by spectrophotometry at defined time points.At the end of experiment,the animals were sacrificed and the samples were collected.The pathological changes of the samples were observed under microscope. Results There were no significant differences in MAP before shock among the five groups.MAP in NS group,PHCL group,PHCM group and PHCH group decreased post-shock,but there were no significant differences in MAP among groups.At T3-T6 time points,MAP in NS group (68.8±3.0),(67.5±5.3),(71.0±3.7),(71.0±2.4) mm Hg( 1 mm Hg=0.133 kPa),respectively was significantly lower than that before shock (94.3±1.7) mm Hg,also lower than that in the Con group (82.8±4.1),(89.8±6.6),(82A±3.3),(94.4±6.2) mm Hg,respectively and the PHCM gronp (83.2±13.6),(82.2±9.0),(83.8±8.9),(85.4±8.1) mm Hg,respectively.There were no significant differences in HR pre-shock and post-shock among the five groups.At T3-T6 time points,HR in the NS group (185±11),(181±7),(164±13),(164±13) hpm,respectively were signifieandy lower than that at T1(276±13) bpm,also significantly (P＜0.05 or P＜0.01 ) lower than that in the Con group and the PHCM group.The plasma DAO activity and D-lactate concentration in the Ns group,PHCL group,PHCM group,PHCH group were significantly increased at T2-T6 time points.Furthermore,at T3-T6 time poinls,the plasma DAO activity (0,393±0,020),(0.586±0.017),(0.844±0.036),(0.568±0.016) μ/ml,respectively and D-lactate concentration(8.292±0.364),(7.539±0.098),(5.991±0.180),(7,108±0.372) mmol/L,respectively in the NS group were significantly higher than that in the PHCL group and PHCH group; the plasma DAO activity and D-lactate concentration in the PHCL group and PHCH group were significantly higher than in the PHCM group (0.111±0.016),(0.302±0.020),(0.501±0.014),(0.183±0.018)μ/ml; (5.664±0.546),(4.609±0.292),(3.310±0,3634),(4.720±0.205)mmol/L(P＜O.05orP＜0.0l).Intestinal mucosal injury in the PHCM group was significantly reduced compared with tbat in the NS group.Conclusions The resuscitation of PHC combinition with Mnormal saline can stabilize hemodynamics in rabbits with traumatic shock.There was a significant protective effect on intestinal mucosal injury induced by traumatic shock,especially in the PHCM group.The plasma DAO activity and D-lactate concentration can be used to assess the extent of intestinal mucosa injury and barrier function. Key words: Traumatic shock; Penehyclidine hydrochloride; Plasma diamine oxidase; D-lactate; Mean arterial blood pressure

Protective effects of penehyclidine hydrochloride on lipopolysaccharide-induced endothelial cells injury

Objective To investigate the effects of penehyclidine hydrochloride ( PHC) on lipopolysaccharide (LPS) -induced endothelial cells injury and its mechanism. Methods ECV-304 was cultured in RPMI1640 in a 5% humidified CO2 atmosphere at 37 ℃. Then cultured cells were used to assess the following treatments: control group, LPS group (1 μg/mL) and PHC group(2 μg/mL). At the end of the experiments, supernatant was collected for determination of lactate dehydrogenase ( LDH), and cells were collected for determination of malondialdehyde (MDA), superoxide dismutase (SOD), and nitric oxide (NO) levels. And extracellular regulated kinasel/2( ERKl/2)and JNK MAPK (mitogen-activated protein kinases, MAPK) protein expressions were determined using Western blot technique. Analysis of variance (ANOVA) was used for statistical analysis to compare values among all groups. A significant difference was presumed for a probability value < 0.05. Results Compared with control group, LDH leakage [(1642 ± 367) U/L vs (169±33)U/L], the contents of MDA[(13. 2 ± 1. 2) nmol/L vs (7. 2 ±0. 8)nmol/mL] and NO levels [(143.2 ± 10.3) μmol/L vs(85.5 ±4.1) μmol/L], expressions of ERK1/2 and JNK were remarkably increased and SOD activities[(41.2 ±2.7) U/mL vs (61. 1 ±2.8) U/mL] were obviously decreased in LPS group. PHC markedly decreased LDH leakage [(392 ±90) U/L], MDA contents [(8. 6 ± 1. 3) nmol/ mL] and NO levels [(92.1 ±6.6) μmol/L], ERK1/2 activation and enhanced SOD activities [(58.0± 3.0) U/mL]. Conclusions PHC could protect endothelial cells against LPS-induced cell injury. The effect of PHC is likely mediated through inhibition of ERK1/2 MAPK activation. Key words: Penehyclidine hydrochloride; Lipopolysaccharide; Endothelial cell; MDA; SOD; NO; ERK1/2; JNK

Preventive effects of ischemic postconditioning and penehyclidine hydrochloride on gastric against ischemia-reperfusion injury in rats

To investigate the protective effects of ischemic postconditioning and penehyclidine hydrochloride on gastric injury induced by ischemia-reperfusion of lower limb in rats. The model of limb ischemia reperfusion injury was used to perform this experiment. One hundred and forty four male Wistar rats weighing 220 - 250 g were randomly divided into 4 groups: group I Control (C), group II Ischemic Reperfusion (IR), group III Ischemic postconditioning (IPO) and group IV penehyclidine hydrochloride (IPHC); C, IR, IPO and IPHC groups has been followed for 0(T(0)), 1(T(1)), 3(T(3)), 6(T(6)), 12(T(12)), or 24(T(24)) perfusion, all the groups were secondary separated into six subgroups as time point and each subgroup contained six rats, respectively. Blood samples from the inferior vena cava were taken for determination of LDH, CK activities and TNF-α, IL-10 content at every time point of reperfusion;the animals were killed at every time point respectively and the gastric were removed for determination of SOD, MPO, XOD and LDH activities, MDA content, and histological examination and the expression of HIF-1α was analyzed. Compared with group C, IR, IPO and IPHC, in serum LDH and CK activities were increased, TNF-α and IL-10 content were increased (P < 0.05 or P < 0.01); and in gastric tissue MPO, XOD and LDH activities were increased and MDA content increased, while SOD activity decreased in group IR, IPO and IPHC (P < 0.05 or P < 0.01); and gastric tissue resulted in significant injury as evidenced by infiltrated of few neutrophils or eosinophils and rare neutrophils between the gastric mucosa or muscularis mucosa and the glands, interstitial vascular dilation hyperemia and small quantity hemorrhage from deep layers of mucosa or interstitial vascular dilation hyperemia, and the expression of HIF-1α was significantly increased (P < 0.01). Compared with group IR, IPO and IPHC in serum LDH and CK activities, TNF-α content decreased while IL-10 content were increased (P < 0.01); and in gastric tissue MDA content, MPO, XOD and LDH activities were decreased, and SOD activity increased in group IPO and IPHC (P < 0.05 or P < 0.01); and the histological injury were milder and the expression of HIF-1α was significantly decreased (P < 0.01). Compared with group IPO, IPHC, in serum LDH activities were makable decreased, CK activities were first increased and then declined, TNF-α content makable declined while IL-10 content were increased (P < 0.05 or P < 0.01), and the histological injury were milder and the expression of HIF-1α was makable decreased (P < 0.01) and in gastric tissue SOD activity were makable increased, MPO activities significantly decreased, MDA content increased at T(3), XOD activities increased after T(12), LDH activities increased at T(3) and declined after T(12) in group IPHC (P < 0.05 or P < 0.01). The combination of ischemia postconditioning and postconditioning with penehyclidine hydrochloride can protect the gastric from ischemia-reperfusion injury induced by ischemia reperfusion of the lower limbs in rats, the main mechanism may be reducing post-ischemic oxidative damage, inflammatory reaction, amelio-rating microcirculatory and cellular energy metabolism et al. Additionally, this study found that the protective effects of penehyclidine hydrochloride on gastric injury induced by ischemia reperfusion of the lower limbs, were better than ischemic postconditioning, and the mechanism might be related to its anti-inflammatory effect, antioxidant action and prevention of cell injury et al.

Protective effects of penehyclidine hydrochloride on liver injury in a rat cardiopulmonary bypass model

Hepatic injury after cardiac surgery is considered to be a consequence of cardiopulmonary bypass (CPB). The aim of this study was to test the hypothesis that penehyclidine hydrochloride (PHC) could attenuate hepatic injury using a rat CPB model. Male Sprague-Dawley rats were randomly divided into six groups (eight per group), including sham-operated control, sham low-dose PHC control (0.6 mg kg), sham high-dose PHC control (2.0 mg kg), vehicle control, low-dose PHC (0.6 mg kg) and high-dose PHC (2.0 mg kg)-treated groups. Blood samples were collected from the femoral artery at the cessation of CPB and the serum levels of the liver enzymes, including alanine aminotransferase (ALT) and aspartate aminotransferase (AST), were determined. The ultrastructure of liver tissue was also examined under an electron microscope. In the sham-operated groups, high-dose PHC and low-dose PHC had no significant impact on the levels of respiratory rate, heart rate, blood pressure, ECG, ALT or AST. Compared with the sham group, the serum levels of ALT and AST were increased significantly in the surgical groups. PHC alleviated all the biochemical and histopathological changes in a dose-dependent manner. The ALT and AST levels in the high-dose PHC-treated groups were significantly lower than those in the vehicle control group. Treatment with penehyclidine hydrochloride could improve liver function during CPB.

Penehyclidine hydrochloride attenuates LPS-induced acute lung injury involvement of NF-κB pathway

To investigate the protective effects of penehyclidine hydrochloride (PHC) in lipopolysaccharide (LPS)-induced acute lung injury (ALI) and the underlying molecular mechanism. ALI was induced by intravenous injection of LPS (5 mg/kg). Male Sprague–Dawley (SD) rats challenged with or without LPS were pretreated with varied doses of PHC 0.5 h before injection of LPS or saline. Blood gas in arterial blood, lung weight gain, bronchoalveolar lavage fluid (BALF), and neutrophils sequestration were examined 6 h after administration of LPS. Pathological changes of lung tissue were measured by light microscopy. Phosphorylation of mitogen-activated protein kinase (MAPK) family and NF-κB were detected by western blot. All animals demonstrated drops in arterial oxygen tension (PaO 2) after LPS application, which were significantly reversed by PHC pretreatment. Administration of PHC reduced lung water gain, bronchoalveolar lavage protein content, infiltration of neutrophils, malondialdehyde (MDA) content, and lactate dehydrogenase (LDH) activity and enhanced superoxide dismutase (SOD) activity. Histopathological study also indicated that PHC treatment markedly attenuated lung histopathological changes, alveolar hemorrhage, and inflammatory cells infiltration with evidence of decreasing of myeloperoxidase (MPO) activity. Furthermore, p38MAPK, ERK, and NF-κB were activated in 6 h after LPS treatment, which could be blunted by PHC, while JNK remained unchanged. These findings confirmed significant protection by PHC against LPS-induced lung vascular leak and inflammation and implicated inhibition of p38MAPK activation signaling a potential role for PHC in the management of ALI.