Protective effect of trans-nerolidol on vascular endothelial cell injury induced by lipopolysaccharides / Efectos protectores del trans-nerolidol en lesiones inducidas por lipopolisacáridos en células endoteliales vasculares

This article aimed to discuss the protection of trans - nerolidol on vascular endothelial cells (ECs) injured by lipopolysac charides. ECs were divided into four groups: normal, model, low and high dose trans - nerolidol treatment groups. The cell survival rate and the contents of NO in the cell culture supernatant were determined. The protein expression and transcript level of pe roxisome proliferator - activated receptor - γ (PPARγ), endothelial nitric oxide synthase (eNOS), and inducible nitric oxide synthase (iNOS) were determined by western blotting and RT - PCR respectively. Compared with the normal group, cell livability, protein e xpression and mRNA transcript level of PPARγ and eNOS decreased, NO contents, protein expression and mRNA transcript tlevel of iNOS increased in model group significantly. Compared with model group, all the changes recovered in different degree in treatmen t groups. Hence, it was concluded that trans - nerolidol can alleviate the ECs injuryby the regulation of iNOS/eNOS through activating PPARγ in a dose - dependent manner

Este artículo tiene como objetivo discutir la protección del trans - nerolidol en las células endoteliales vasculares (CE) dañadas por lipopolisacáridos. Las CE se di vidieron en cuatro grupos: normal, modelo, grupos de tratamiento con trans - nerolidol de baja y alta dosis. Se determinó la tasa de supervivencia de las células y los contenidos de óxido nítrico (NO) en el sobrenadante del cultivo celular. La expresión de p roteínas y el nivel de transcripción del receptor activado por proliferadores de peroxisomas - γ (PPARγ), el óxido nítrico sint et asa endotelial (eNOS) y el óxido nítrico sint et asa inducible (iNOS) se determinaron mediante western blot y RT - PCR, respectivamen te. En comparación con el grupo normal, la viabilidad celular, la expresión de proteínas y el nivel de transcripción de PPARγ y eNOS disminuyeron, los contenidos de NO, la expresión de proteínas y el nivel de transcripción de iNOS aumentaron significativam ente en el grupo modelo. En comparación con el grupo modelo, todos los cambios se recuperaron en diferentes grados en los grupos de tratamiento. Por lo tanto, se concluyó que el trans - nerolidol puede aliviar el daño en las CE regulando iNOS/eNOS a través d e la activación de PPARγ de manera dependiente de la dosis.

Role of endothelial nitric oxide synthase in exercise preconditioning-induced improvement of myocardial ischemia-reperfusion injury / 中国组织工程研究

BACKGROUND:Exercise is an effective strategy to prevent and treat various cardiovascular diseases and protect the heart from ischemia-reperfusion injury.Its mechanism of action needs to be studied in depth. OBJECTIVE:To observe the effect of aerobic exercise preconditioning on myocardial ischemia-reperfusion injury and to explore the effect of endothelial nitric oxide synthase(eNOS)activation(including coupling and phosphorylation). METHODS:Eighty adult Wistar rats were randomly divided into sedentary(n=40)and exercise(n=40)groups.The rats in the exercise group were subjected to aerobic exercise for 8 weeks while those in the sedentary group were quietly fed and caged.After 8 weeks of intervention,three experiments were performed.(1)Experiment 1:After the last training,cardiac function,cardiac nitric oxide metabolite content and cardiac eNOS,phosphorylated eNOS-S1177,eNOS dimer and eNOS monomer protein expression levels were detected.(2)Experiment 2:Rats were divided into sedentary control group,exercise control group,sedentary+eNOS inhibitor group,exercise+eNOS inhibitor group,all of which were subjected to an in vitro myocardial ischemia-reperfusion injury experiment.eNOS inhibitor was continuously infused into the sedentary+eNOS inhibitor group and exercise+eNOS inhibitor group 10 minutes before reperfusion,and cardiac function and myocardial infarction area were detected 3 hours after reperfusion.(3)Experiment 3:Rats were divided into sedentary control group,exercise control group,sedentary+eNOS coupler group and exercise+eNOS coupler group,all of which were subjected to an in vitro myocardial ischemia-reperfusion injury experiment.The rats in the sedentary+eNOS coupler group and exercise+eNOS coupler group were treated with eNOS coupler.Myocardial infarct area,cardiac nitric oxide metabolite content,cardiac protein expression of eNOS,phosphorylated eNOS-S1177,eNOS dimer,eNOS monomer and 3-nitrotyrosine were detected 3 hours after reperfusion.The phosphorylated eNOS-S1177/eNOS ratio reflected the phosphorylated/dephosphorylated level of eNOS and eNOS dimer/monomer ratio reflected eNOS coupling/uncoupling level. RESULTS AND CONCLUSION:Experiment 1:Compared with the sedentary group,the exercise group had increased cardiac output and left ventricular ejection fraction(P<0.05),increased nitrite and S-nitrosothiol contents(P<0.05),upregulated phosphorylated eNOS-S1177,eNOS protein expression and phosphorylated eNOS-S1177/eNOS ratio(P<0.05),eNOS dimer protein expression and eNOS dimer/monomer ratios were elevated(P<0.05).Experiment 2:Compared with the sedentary control group,left ventricular development pressure increased(P<0.05)and myocardial infarct area decreased(P<0.05)in the exercise control group.Compared with the exercise control group,left ventricular development pressure decreased(P<0.05)and myocardial infarct area increased(P<0.05)in the exercise+eNOS inhibitor group.Experiment 3:Compared with the sedentary control group,the exercise control group had increased left ventricular developmental pressure(P<0.05),decreased myocardial infarct area(P<0.05),decreased phosphorylated eNOS-S1177/eNOS ratio(P<0.05),decreased eNOS dimer/monomer ratio(P<0.05),increased S-nitrosothiol content(P<0.05),and decreased 3-nitrotyrosine protein expression(P<0.05).Compared with the exercise control group,the exercise+eNOS coupler group had decreased left ventricular developmental pressure(P<0.05),increased myocardial infarct area(P<0.05),increased phosphorylated eNOS-S1177/eNOS ratio(P<0.05),increased eNOS dimer/monomer ratio(P<0.05),and elevated 3-nitro tyrosine protein expression(P<0.05).To conclude,aerobic exercise preconditioning could induce cardioprotection,which is related to uncoupling and dephosphorylation of eNOS during cardiac ischemia-reperfusion,thereby inhibiting the excessive production of nitric oxide and reducing nitro-oxidative stress.

Placental expression of striatin & endothelial nitric oxide synthase in women with & without pre-eclampsia

Background & objectives: Striatin is a multi-domain scaffolding protein essential for activating endothelial nitric oxide synthase (eNOS). However, its role in pre-eclampsia remains use explored. Hence, this study aimed to investigate the association between striatin and eNOS in regulating nitric oxide (NO) production in the placenta of women with and without pre-eclampsia. Methods: Forty pregnant women each without (controls) and with pre-eclampsia (cases) were enrolled in the study. Blood striatin and NO concentrations were detected by the ELISA. Protein expression of striatin, phosphorylated eNOS (peNOS), inducible NOS (iNOS) and phosphorylated NF-?B were measured in the placental tissues by Western blot. Twenty four hour urinary protein and serum urea, uric acid and creatinine were analyzed as an autoanalyzer. Placental histology was analyzed by haematoxylin and eosin staining. Results: Compared to normotensive pregnant women, the levels of serum NO and striatin were decreased in pre-eclamptic women. The protein expression of striatin and peNOS was significantly reduced (P<0.05) while p65NF-?B and iNOS were upregulated considerably (P<0.05) in the placenta of cases compared to controls. Interpretation & conclusions: Our results show for the first time that decreased striatin expression was associated with decreased peNOS protein expression in the placental tissue of pre-eclamptic women. Interestingly, no significant difference was found in blood striatin or NO levels between controls and cases. Thus, therapies that improve placental striatin expression are attractive possibilities, both for prevention as well as treatment of endothelial dysfunction in pre-eclampsia.

Oxymatrine improves palmitic acid-induced vascular endothelial cell injury through Akt-eNOS-NO signaling pathway / 中国药理学通报

Aim To investigate the protective effect of oxymatrine (OMT) on vascular endothelial cell injury induced by palmitic acid ( PA) and its mechanism. Methods Cell viability was detected by MTT assay. Cell apoptosis was detected by flow cytometry. The levels of oxygen species ( ROS) in cells, and lactate de-hydrogenase, malondialdehyde (MDA), superoxide dismutase (SOD) , glutathione peroxidase (GSH-PX) and nitric oxide ( NO) in cell culture medium were detected by ELISA. The protein expressions of bcl-2, bax, caspase-3, Akt and eNOS in HUVECs were detected by Western blot. Results OMT significantly inhibited PA-induced decrease in cell viability and increase in level of LDH in HUVECs. OMT also significantly inhibited PA-induced increase in cell apoptosis, and up-regulated the protein expression ratio of bcl-2/ bax and down-regulated the protein expression of caspase-3. In addition, OMT reduced the levels of ROS and MDA, and increased the levels of SOD, GSH-Px and NO in cell-culture medium treated with PA. Furthermore, OMT increased the protein phospho-rylation of Akt and eNOS in injured cells. Conclusion OMT ameliorates PA-induced vascular endothelial cell injury through Akt-eNOS-NO signaling pathway.

Shear stress regulateing the phosphorylation of endothelial nitric oxide synthase Ser633 and Ser1177 in human umbilical vein endothelial cells through Pim1 / Akt pathway / 解剖学报

[Abstract] Objective To explore the effect of different modes of blood flow shear stress on the Pim1 expression in human umbilical vein endothelial cells (HUVECs) and the regulation of phosphorylation at Ser1177 and Ser633 of endothelial nitric oxide synthase (eNOS). Methods HUVECs were isolated from fresh human umbilical cord. The parallel plate flow chamber system was used to load 15 dyn/ cm

Phosphorylated PKM2 regulates endothelium-dependent vasodilation in diabetes / 中南大学学报(医学版)

OBJECTIVES@#Endothelium-dependent vasodilation dysfunction is the pathological basis of diabetic macroangiopathy. The utilization and adaptation of endothelial cells to high glucose determine the functional status of endothelial cells. Glycolysis pathway is the major energy source for endothelial cells. Abnormal glycolysis plays an important role in endothelium-dependent vasodilation dysfunction induced by high glucose. Pyruvate kinase isozyme type M2 (PKM2) is one of key enzymes in glycolysis pathway, phosphorylation of PKM2 can reduce the activity of pyruvate kinase and affect the glycolysis process of glucose. TEPP-46 can stabilize PKM2 in its tetramer form, reducing its dimer formation and phosphorylation. Using TEPP-46 as a tool drug to inhibit PKM2 phosphorylation, this study aims to explore the impact and potential mechanism of phosphorylated PKM2 (p-PKM2) on endothelial dependent vasodilation function in high glucose, and to provide a theoretical basis for finding new intervention targets for diabetic macroangiopathy.@*METHODS@#The mice were divided into 3 groups: a wild-type (WT) group (a control group, C57BL/6 mice) and a db/db group (a diabetic group, db/db mice), which were treated with the sodium carboxymethyl cellulose solution (solvent) by gavage once a day, and a TEPP-46 group (a treatment group, db/db mice+TEPP-46), which was gavaged with TEPP-46 (30 mg/kg) and sodium carboxymethyl cellulose solution once a day. After 12 weeks of treatment, the levels of p-PKM2 and PKM2 protein in thoracic aortas, plasma nitric oxide (NO) level and endothelium-dependent vasodilation function of thoracic aortas were detected. High glucose (30 mmol/L) with or without TEPP-46 (10 μmol/L), mannitol incubating human umbilical vein endothelial cells (HUVECs) for 72 hours, respectively. The level of NO in supernatant, the content of NO in cells, and the levels of p-PKM2 and PKM2 protein were detected. Finally, the effect of TEPP-46 on endothelial nitric oxide synthase (eNOS) phosphorylation was detected at the cellular and animal levels.@*RESULTS@#Compared with the control group, the levels of p-PKM2 in thoracic aortas of the diabetic group increased (P<0.05). The responsiveness of thoracic aortas in the diabetic group to acetylcholine (ACh) was 47% lower than that in the control group (P<0.05), and that in TEPP-46 treatment group was 28% higher than that in the diabetic group (P<0.05), while there was no statistically significant difference in the responsiveness of thoracic aortas to sodium nitroprusside (SNP). Compared with the control group, the plasma NO level of mice decreased in the diabetic group, while compared with the diabetic group, the phosphorylation of PKM2 in thoracic aortas decreased and the plasma NO level increased in the TEPP-46 group (both P<0.05). High glucose instead of mannitol induced the increase of PKM2 phosphorylation in HUVECs and reduced the level of NO in supernatant (both P<0.05). HUVECs incubated with TEPP-46 and high glucose reversed the reduction of NO production and secretion induced by high glucose while inhibiting PKM2 phosphorylation (both P<0.05). At the cellular and animal levels, TEPP-46 reversed the decrease of eNOS (ser1177) phosphorylation induced by high glucose (both P<0.05).@*CONCLUSIONS@#p-PKM2 may be involved in the process of endothelium-dependent vasodilation dysfunction in Type 2 diabetes by inhibiting p-eNOS (ser1177)/NO pathway.

Mechanism of miR-29a affecting eNOS protein expression and participating in vascular endothelial dysfunction / 中国现代医生

Objective To explore the inhibitory effect of miR-29a on in vitro angiogenesis of vascular endothelial cells by targeting cells with glutaminase,and preliminarily explore the relevant mechanisms of inflammatory factors inducing vascular endothelial dysfunction.Methods Human umbilical vein endothelial cells were selected as research cell lines,and the cell lines were intervened by recombinant human tumor necrosis factor-α(TNF-α)and serum amyloid A(SAA)respectively.The expression changes of miRNA were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).The expression changes of endothelial nitric oxide synthase(eNOS)were detected by Western blot.Finally,the vascular protective drug salvia miltiorrhiza was used to reverse verify the expression changes of miR-29a and eNOS.Results The addition of TNF-α and SAA significantly increased the expression level of miR-29a compared with control group(P<0.05).qRT-PCR showed that the higher the level of inflammatory factors,the higher the expression level of cell line miR-29a.With 1.0ng/ml TNF-α and SAA as a constant dose,it was found that the expression level of miR-29a increased gradually with time.Western blot showed that the addition of TNF-α and SAA would decrease the level of eNOS protein in the cell line.The expression level of miR-29a decreased with the increase of salvia miltiorrhiza concentration,and decreased with time at constant salvia miltiorrhiza concentration.After adding salvia miltiorrhiza,the expression of eNOS protein increased obviously.Conclusion Inflammatory factors TNF-α and SAA can participate in the process of vascular endothelial injury by inducing the expression of miR-29a,and eNOS protein is widely involved in this process.The application of vasoprotective drugs can inhibit the over-expression of miR-29a to some extent,reduce the expression of eNOS protein,and thus play a role in protecting blood vessels.

Effect of Xileisan temperature-sensitive gels on endothelial nitric oxide synthase, vascular endothelial growth factor A and tumor necrosis factor-α expression in rats with bleeding internal hemorrhoids / 预防医学

Objective@#To investigate the effect of Xileisan temperature-sensitive gels on endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor A (VEGF-A) and tumor necrosis factor-α (TNF-α) expression in rats with bleeding internal hemorrhoids, so as to provide insights into the illustration of the pathogenesis of internal hemorrhoid hemorrhage. @*Methods@#Thirty six-week-old SPF-graded rats of the SD strain were randomly divided into the normal group, model group and Xileisan temperature-sensitive gel group, of 10 rats in each group (half male and half female). Cotton balls were soaked with 0.16 mL of croton oil mixture and then inserted into the anus of rats in the model group and Xileisan temperature-sensitive gel group for 10 s. After 6 h when the rectal mucosa tissues presented remarkable swelling, the perianal mucosa was rubbed repeatedly with a rough glass rod until the glass rod was bloody. Following successful modeling, rats in the Xileisan temperature-sensitive gel group was given rectal administration of Xileisan temperature-sensitive gel at a dose of 0.5 mL/d, while animals in the normal group and model group were given rectal administration of the blank gel at the same dose. Following administration for 7 successive days, rats were sacrificed, and the hemorrhoids tissues were collected for pathological examinations. The eNOS, VEGF-A and TNF-α expression was determined using immunohistochemistry and compared among groups.@*Results@#Compared with the normal group, the rat hemorrhoids mucosa showed inflammatory changes in the model group, with submucosal congestion and edema, blood vessel congestion and dilation, and visible new blood vessels, and remarkable improvements were seen in the hemorrhoid mucosal inflammation in the Xileisan temperature-sensitive gel group. There were significant differences in the integrated option density (IOD) of eNOS and VEGF-A expression in rat hemorrhoids tissues among the three groups (P<0.05), and no gender-specific differences were seen (P>0.05). The IOD values of eNOS (45.84±13.66) and VEGF-A expression (45.89±9.06) were higher in rat hemorrhoids tissues in the model group than in the normal group (23.11±5.64 and 27.91±11.65) and the Xileisan temperature-sensitive gel group (27.41±8.89 and 33.44±6.20) (P<0.05), while no significant differences were detected in the IOD of TNF-α expression in rat hemorrhoids tissues among the three groups (P>0.05).@*Conclusion@#Xileisan temperature-sensitive gel may alleviate inflammation and internal hemorrhoids hemorrhage through inhibiting eNOS and VEGF-A expression in rat hemorrhoids tissues.

Relationship between the G894T polymorphism of endothelial nitric oxide synthase gene and lipid metabolism among hypertensive disorder complicating pregnancy patients / 解剖学报

Objective To investigate the relationship between the G894T polymorphism of the endothelial nitric oxide synthase (eNOS) gene and the lipid metabolism in patients with irypertensive disorder complicating pregnancy (HDCP). Methods The 528 cases of HDCP patients admitted to the Xingtai Third Hospital from January 2016 to January 2020 were selected as the research objects, and 128 normal pregnant women during the same period were selected as the control group. The fasting peripheral venous blood of all stud)' subjects in the early morning was collected, and blood lipid indexes, cystatin C (CysC) and uric acid levels and other biochemical index levels were measured. According to the blood lipid level, it was divided into normal blood lipid group and dyslipidemia group. The dyslipidemia group included 4 subgroups [ hyper triglyceride (TG) blood group (T G

Involvement of ET-1/eNOS in the ameliorating effect of electroacupuncture on cardiac dysfunction in rats with spontaneously hypertensive / 中国针灸

OBJECTIVE@#To observe the effect of electroacupuncture (EA) at "Neiguan" (PC 6) on cardiac function of ventriculus sinister in rats with spontaneously hypertensive (SHR), and to explore the mediation effect of endothelin-1 (ET-1)/endothelial nitric oxide synthase (eNOS).@*METHODS@#Six 12-week-old male Wistar Kyoto (WKY) rats were taken as the normal group. Eighteen 12-week-old SHR were randomly divided into a model group, an EA group and a sham EA group, 6 rats in each group. The rats in the EA group were treated with EA (disperse-dense wave, 2 Hz/15 Hz in frequency, 1 mA in current intensity) at "Neiguan" (PC 6), 30 min each time, once a day for 8 weeks. The rats in the sham EA group were treated with superficial needling at "Neiguan" (PC 6) with no electrical stimulation applied. After treatment, the left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were tested by echocardiographic analysis. The left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), heart rate (HR), the maximum rate of increase/decrease of left ventricular pressure (±dp/dtmax) were detected. The serum content of ET-1 was detected by ELISA. Western blot was used to evaluate the expression of ETAR, eNOS in myocardial tissue of left ventricular.@*RESULTS@#Compared with the normal group, LVEF, LVFS, +dp/dtmax/LVSP and -dp/dtmax/LVSP were decreased (P<0.01, P<0.05), while LVSP, LVEDP, +dp/dtmax and -dp/dtmax were increased (P<0.01) in the model group. Compared with the model group, LVEF, LVFS, +dp/dtmax/LVSP and -dp/dtmax/LVSP were increased (P<0.01, P<0.05), and LVSP and LVEDP were decreased (P<0.01) in the EA group. Compared with the normal group, the serum content of ET-1 and the expression of ETAR in myocardial tissue were increased (P<0.01), whereas expression of eNOS was decreased (P<0.01) in the model group. Compared with the model group, the serum content of ET-1 and the expression of ETAR in myocardial tissue were decreased (P<0.05), whereas expression of eNOS was increased (P<0.05) in the EA group.@*CONCLUSION@#EA intervention may alleviate hypertensive cardiac function damage by up-regulating the expression of eNOS protein in myocardial tissue, down-regulating the serum content of ET-1 and the expression of ETAR protein in myocardial tissue.

Serum NOS3 and GDNF levels in patients with arteriosclerotic cerebral infarction and their clinical significance / 中风与神经疾病杂志

@#Objective　To investigate the expression differences of serum endothelial nitric oxide synthase (NOS3) and glial cell-derived neurotrophic factor (GDNF) in patients with arteriosclerotic cerebral infarction and their role in the prognosis of patients.Methods　A total of 126 patients with atherosclerotic cerebral infarction admitted to our hospital from January 2021 to January 2022 and 130 healthy people in the same period were selected.Patients were divided into groups according to infarct volume,dysfunction and prognosis,and serum NOS3 and GDNF levels were compared between different groups.ROC curve was used to analyze the predictive value of NOS3 and GDNF for poor prognosis.Results　The proportion of diabetes and hypertension in patients with atherosclerotic cerebral infarction increased,and the serum levels of NOS3 and GDNF decreased (P<0.05).There was no significant difference in the levels of NOS3 and GDNF among patients with different infarct volumes (P>0.05).The levels of NOS3 and GDNF in mild dysfunction group,moderate dysfunction group and severe dysfunction group decreased successively (P<0.05).The AUC of NOS3 and GDNF levels in predicting poor prognosis were 0.858 and 0.779,respectively.Conclusion　The serum levels of NOS3 and GDNF in patients with atherosclerotic cerebral infarction are decreased,and their levels are related to the degree of neurological impairment and prognosis of patients,which is expected to be a biological indicator to evaluate the prognosis of patients.

Nitric oxide regulates adhesiveness, invasiveness, and migration of anoikis-resistant endothelial cells

Anoikis is a type of apoptosis that occurs in response to the loss of adhesion to the extracellular matrix (ECM). Anoikis resistance is a critical mechanism in cancer and contributes to tumor metastasis. Nitric oxide (NO) is frequently upregulated in the tumor area and is considered an important player in cancer metastasis. The aim of this study was to evaluate the effect of NO on adhesiveness, invasiveness, and migration of anoikis-resistant endothelial cells. Here, we report that anoikis-resistant endothelial cells overexpress endothelial nitric oxide synthase. The inhibition of NO release in anoikis-resistant endothelial cells was able to decrease adhesiveness to fibronectin, laminin, and collagen IV. This was accompanied by an increase in cell invasiveness and migration. Furthermore, anoikis-resistant cell lines displayed a decrease in fibronectin and collagen IV protein expression after L-NAME treatment. These alterations in adhesiveness and invasiveness were the consequence of MMP-2 up-regulation observed after NO release inhibition. The decrease in NO levels was able to down-regulate the activating transcription factor 3 (ATF3) protein expression. ATF3 represses MMP-2 gene expression by antagonizing p53-dependent trans-activation of the MMP-2 promoter. We speculate that the increased release of NO by anoikis-resistant endothelial cells acted as a response to restrict the MMP-2 action, interfering in MMP-2 gene expression via ATF3 regulation. The up-regulation of nitric oxide by anoikis-resistant endothelial cells is an important response to restrict tumorigenic behavior. Without this mechanism, invasiveness and migration potential would be even higher, as shown after L-NAME treatment.

Human umbilical cord Wharton's jelly-derived mesenchymal stem cell transplantation could improve diabetic intracavernosal pressure / 亚洲男科学杂志(英文版)

Mesenchymal stem cells (MSCs) secrete various cytokines with angiogenic and neuroprotective effects. This study aimed to assess the effects of human umbilical cord Wharton's jelly-derived MSCs (hWJ-MSCs) on diabetes-related intracavernosal pressure (ICP) impairment in rats. hWJ-MSCs were isolated from human umbilical cord Wharton's jelly and transplanted into the corpus cavernosum of streptozotocin (STZ)-induced diabetic rats by unilateral injection. The erectile function was evaluated at 4 weeks, as well as the expression levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), endothelial nitric oxide synthase (eNOS), and insulin-like growth factor 1 (IGF1). STZ-induced diabetic rats showed impaired ICP, which was significantly improved by hWJ-MSC treatment. VEGF, eNOS, IGF1, and bFGF expression levels were higher in hWJ-MSC injection sites than those in control ones in STZ-induced diabetic rats. These results suggest that hWJ-MSC transplantation might improve diabetic erectile dysfunction through increased production of paracrine growth factors, highlighting a novel potential therapeutic option for erectile dysfunction.

Fasudil alleviates lung injury via targeting Rhoa/ROCK signal pathway in septic mice / 中华急诊医学杂志

Objective:To determine the protective effect of fasudil on acute lung injury in septic mice.Methods:Forty-five 4-6-week-old male C57BL mice were randomly(random number) assigned to three groups ( n=15 each group): control group, lipopolysaccharide (LPS) group and Fasudil intervention group (FAS+LPS). Acute lung injury model of septic mice was established with an intraperitoneal injection and intratracheal infusion of LPS. The mice in the FAS+LPS group were injected with fasudil hydrochloride intraperitoneally 30 min before intraperitoneal LPS injection and 1 h after intratracheal LPS infusion, respectively. All mice were sacrificed at 4 h after modeling, and lung tissues were collected. Hematoxylin-eosin staining was preformed to observe the morphological changes in the lung tissue. The wet /dry weight (W/D) ratio, malondialdehyde (MDA) content and the activity of myeloperoxidase (MPO) in the lung tissues were detected. Caspase-3 expression was examined by immunohistochemical (IHC) staining. Western blot was employed to detect the expression of RhoA, ROCK1, endothelial nitric oxide synthase (eNOS), and p-eNOS. Results:Inflammatory cell infiltration and erythrocyte exudation were significantly reduced, and the degree of interstitial oedema and derangement of alveolar structure appeared in a decreasing degree after FAS intervention. Compared with the LPS group, the W/D ratio, MDA content, MPO activity and the expression of Caspase-3 in the FAS+LPS group were significantly reduced (all P<0.01). Meanwhile, the expression of RhoA and ROCK1 of the LPS group were obviously higher than those in the control group ( P<0.05), and p-eNOS was obviously lower than that in the control group ( P<0.05). Furthermore, the expression of RhoA and ROCK1 of the FAS+LPS group were obviously lower than those in the LPS group, and p-eNOS was obviously higher than that in the LPS group. There was no significant difference on the expression of eNOS among the three groups. Conclusions:Fasudil can alleviate the degree of inflammatory cell infiltration, reduce apoptosis in lung tissue, inhibit the RhoA/ROCK1 signaling activity, and promote the phosphorylation expression of eNOS in septic mice.

Role of eNOS/PKG-1 pathway in L-Arginine against right ventricular remodeling induced by monocrotaline in rats / 中国药理学通报

Aim To investigate the role of eNOS/PKG- 1 pathway in L-arginine (L-arg) intervention in right ventricular remodeling induced by monocrotaline (MCT) in Sprague Dawley (SD) rats with the aid of the tool medicine L-NAME. Methods Twenty SD rats were randomly divided into control group, MCT group, L-Arg group and L-Arg + L-NAME group. The general condition of rats was observed; the right ventricular pressure of rats was measured by right heart catheterization; the rats and the right ventricle were weighed and the right ventricular mass index was calculated; the morphological changes of the right ventricular were observed by H&E staining; the protein expressions of cTnl, eNOS and PKG-1 were detected by Western blot in the right ventricular. Results Compared with control group, right ventricular max pressure and right ventricular mass index significantly increased ( P < 05) ; the weight of rats in MCT group was significantly reduced ( P < 0. 05); the right ventricular myocytes were hypertrophic, disordered and infiltrated with inflammatory cells; the protein expression of cTnl was obviously up-regulated ( P < 0. 05 ) ; the protein expressions of eNOS and PKG-1 were significantly down- regulated ( P < 0. 05 ) . L-arg could significantly improve the above changes ( P < 0. 05 ). However, the effects of L-arg were inhibited by eNOS inhibitor L- NAME. Conclusions L-arg can improve the right ventricular remodeling in rats induced by MCT, and the mechanism may be related to the up-regulation of the protein levels of eNOS and PKG-1.

Effect of Danggui Buxuetang on Functional Activity of Endothelial Progenitor Cells Exposed to Shear Stress / 中国实验方剂学杂志

Objective:To investigate the effect of Danggui Buxuetang（DGBX）on the functional activity of rat endothelial progenitor cells（EPCs）exposed to different luminar shear stress （SS）. Method:EPCs isolated from rat bone marrow were incubated on a parallel plate flow chamber at a steady SS of 0， 0.12， 1.2， 2.4 Pa for 6 h，then the cells exposed to different SS were randomly divided into 8 groups： control group （perfused with serum free medium），simvastatin group（0.1 μmol·L^-1simvastatin），3 DGBX groups（low，medium，high-dose DGBX）and 3 inhibitor groups（3 DGBX groups with LY294002）. After 12 h，the samples were collected for the detection of cell proliferation ，migration，tubule formation ，the secretion of nitric oxide （NO） ，and the expressions of endothelial nitric oxide synthase（eNOS） mRNA and protein kinase B（Akt），respectively. Result:Compared with the control group，simvastatin and DGBX（high-dose）could both promote the functional activities and NO secretion，and up-regulate the expressions of eNOS mRNA and Akt protein in EPCs exposed to different SS（<italic>P</italic><0.05），while DGBX（mid-dose）could do these only at 0 Pa. However，LY294002 could inhibit all effects of DGBX on EPCs. Conclusion:SS seems to play an important role in the effect of DGBX on EPCs，and DGBX could promote the functional activity of EPCs exposed to SS by up-regulating the expressions of NO/eNOS/Akt.

Sesamin and vitamin E ameliorate aortic endothelial dysfunction rats induced by D-galactose and aluminum trichloride via regula?tion of endothelial nitric oxide synthase and NAD(P)H oxidase 4 / 中国药理学与毒理学杂志

OBJECTIVE To observe the protective effect of sesamin (Ses) and vitamin E (Vit E) against aortic endothelial dysfunction in rats induced by D-galactose (D-gal) and aluminum trichloride (AlCl3), and explore its conceivable mechanisms. METHODS A model of aortic endothelial dysfunction rats was established by D-gal (180 mg · kg-1, ip) combined with AlCl3 (15 mg · kg-1, ig) for 84 d. Model rats were randomly divided into model, model+Vit E 10 mg·kg-1, model+Ses 160 mg·kg-1, and model+Ses 160 mg · kg-1+Vit E 10 mg · kg-1 groups. After 70 d of treatment with Ses and Vit E, systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean blood pressure (MBP) were measured by tail cuff. The rats were anesthetized by sodium pentobarbital (30 mg·kg-1, ip). Thoracic aortas from the rats were removed and divided into two parts (3 mm in length). The relaxation of the aortic ring induced by acetylcholine (ACh) and sodium nitroprusside was measured. The primary pathologic changes in the aorta were observed by HE staining. Total antioxidant capacity (T-AOC), hydrogen peroxide (H2O2) and nitric oxide (NO) in serum were measured by colorimetric analysis. The expression of endothelial nitric oxide synthase (eNOS) positive cells in the aorta were measured by immunohistochemistry. The expres?sions of eNOS and NAD(P)H oxidase 4 (NOX4) protein in the aortal were detected by Western blotting. RESULTS Compared with the model group, the relaxation response with increase in ACh concentra?tion (1×10-7-1×10-4 mol·L-1) was enhanced (P<0.01) in model+Ses+Vit E, SBP, DBP and MBP decreased (P<0.01), the serum T-AOC and NO level were increased (P<0.01), the serum H2O2 levels were reduced (P<0.01), the eNOS expression was increased (P<0.01) and NOX4 expression was reduced (P<0.01) in each treatment group. Compared with model+Ses, the SBP, DBP and MBP were lower (P<0.01 or P<0.05), the serum H2O2 level was lower (P<0.01), the serum NO level was increased (P<0.05), the eNOS expression level was higher (P<0.01) and the NOX4 expression level was reduced (P<0.05) in model+Ses+Vit E. Compared with the model+Vit E, the serum T-AOC and NO levels were increased (P<0.05), the serum H2O2 level was lower (P<0.01), eNOS expression was increased (P<0.01) and NOX4 expression was reduced (P<0.05) in model+Ses+Vit E group. CONCLUSION Ses and Vit E can ameliorate aortic endothelial dysfunction of rats induced by D-gal and AlCl3 via the regulation of eNOS and NOX4.

Effect of Sanhuang Xiexintang in Inhibiting 7-Ketocholesterol-induced Endothelial Disfunctional by NIRP3 Inflammasome Pathway / 中国实验方剂学杂志

<b>Objective::To study whether Sanhuang Xiexintang (SHXXT) can restore endothelial function by inhibiting the activation of NOD-like receptor protein 3 (NIRP3) induced by 7-ketocholesterol (7-keto) in vascular endothelial cells. <b>Method::The aortic rings of mice were cultured in normal group, model (7-keto) group, SHXXT groups (1%, 2% and 5% drug-containing serum). Vasodilation function of mice was observed. Microvascular endothelial cells were cultured according to the above experimental groups, and NIRP3 inhibitor isoglycyrrhizin (ISO) group, was also set. Western blot was used to detect the expressions of endothelial nitric oxide synthase (eNOS), NIRP3, cysteinyl aspartate specific proteinase-1 (Caspase-1), interleukin-1<italic>β</italic> (IL-1<italic>β</italic>) protein. In addition, nitric oxide (NO) quantitative kit was used to detect the concentration of NO. <b>Result::Compared with the normal group, the endothelium-dependent vasodilation function of vascular rings was significantly reduced in model group (<italic>P</italic><0.01), and the drug group significantly restored the endothelium-dependent vasodilation function in a concentration-dependent manner (<italic>P</italic><0.05, <italic>P</italic><0.01). Meanwhile, microvascular endothelial cells were also studied. Compared with the normal group, the content of eNOS protein in the model group decreased (<italic>P</italic><0.05), while the concentration of NO decreased significantly (<italic>P</italic><0.01). After treatment with SHXXT serum, eNOS and NO could be restored, with significant differences in the concentration of NO with 5% (<italic>P</italic><0.05) and 10% (<italic>P</italic><0.01) SHXXT serum. At the same time, the expressions of NIRP3 (<italic>P</italic><0.05), cle-Caspase-1 activation (<italic>P</italic><0.01) and IL-1<italic>β</italic> production (<italic>P</italic><0.01) in endothelium were significantly increased under 7-keto stimulation, and the SHXXT serum could significantly inhibit the expression and activation of relevant proteins. Subsequently, endothelial cells were treated with NIRP3 inhibitor ISO. Compared with the model group, eNOS expression increased, and NO concentration increased significantly (<italic>P</italic><0.01) after treatment with ISO, but ISO had no synergistic effect on SHXXT serum. <b>Conclusion::SHXXT can improve endothelium-dependent vascular dysfunction induced by 7-keto, which is achieved by NO signaling pathway mediated by inhibiting the activation of endothelial NIRP3-related proteins.

Protective effect of Bifidobacterium on endothelial nitric oxide synthase in neonatal necrotizing enterocolitis model / 解放军医学杂志

Objective: To investigate the effect of Bifidobacterium on endothelial nitric oxide synthase and its potential protective mechanism in neonatal necrotizing enterocolitis using an induced mouse model. Methods: Sixty 8-day newborn C57BL/6 mice were randomized into three groups (n=20): control group, necrotizing enterocolitis group (NEC) and Bifidobacterium intervention group (NEC+BIF). Samples were collected 3 days after NEC modeling. HE staining pathological score was used to evaluate the modeling effect; the level of endothelial nitric oxide synthase (eNOS) in the intestine was detected with Western blotting; the eNOS activity, nitric oxide (NO) and superoxide (O2•) content were measured by Colorimetry. Real-time quantitative PCR was used to detect the mRNA level of interleukin-6 (IL-6) and interleukin-1β (IL-1β); the protein expression of IL-6 and IL-1β was assessed by ELISA. Results: No significant difference was observed in the protein levels of eNOS between the three groups. Compared with control group, in NEC group, the eNOS activity and NO content were significantly decreased; the O2 content was substantially increased; both IL-6 and IL-1β were dramatically increased (P0.05). Conclusion: Bifidobacterium can alleviate the severity of intestinal damage in NEC mouse model, the mechanism may be related to the enhancement of eNOS activity.

Remifentanil alleviates renal ischemia/reperfusion injury by activating PI3K/Akt/eNOS pathway / 第二军医大学学报

Objective: To explore the effect and mechanism of intraoperative remifentanil (RF) on renal ischemia/reperfusion (I/R) injury. Methods: Fifty male C57/BL mice were randomly divided into 5 groups: sham group, I/R group, I/R+LY294002 (a phosphatidylinositol 3-kinase [PI3K] inhibitor) group, I/RRF group and I/R + RF+LY294002 group, with 10 mice in each group. Venous blood or renal tissue samples were collected from the mice of each group 6 h after I/R operation. The levels of blood urea nitrogen (BUN) and serum creatinine (SCr) were detected using automatic biochemical analyzer. The expression levels of PI3K/protein kinase B (Akt)/endothelial nitric oxide synthase (eNOS) pathway-related proteins in renal tissues of mice were detected using Western blotting. The aggregation of inflammatory cells was observed by H-E staining. The expression levels of tumor necrosis factor α (TNF-α), interleukin (IL)-1β, IL-6 and IL-10 in renal tissues of mice were detected by enzyme-linked immunosorbent assay. The mRNA expression levels of anti-apoptotic factor Bcl2 and apoptotic factor Caspase-3 in renal tissues were determined by real-time quantitative PCR. Results: Compared with the sham group, the BUN and SCr levels in venous blood were increased in the I/R group, the PI3K expression, phosphorylated-Akt (p-Akt)/Akt ratio and phosphorylated-eNOS (p-eNOS)/eNOS ratio in renal tissues were decreased, the release levels of inflammatory factors (TNF-α, IL-1β, IL-6 and IL-10) were increased, Bcl2 mRNA expression was decreased, and Caspase-3 mRNA expression was increased; and the differences were significant (all P < 0.05). The mice of the I/R group had increased inflammatory cell recruitment in renal tissues. After RF treatment, the mice of the I/R + RF group had decreased levels of BUN and SCr in venous blood, increased PI3K expression, p-Akt/Akt ratio and p-eNOS/eNOS ratio in renal tissues, decreased release levels of inflammatory factors (TNF-α, IL-1β, IL-6 and IL-10), increased Bcl2 mRNA expression, and decreased Caspase-3 mRNA expression; and the differences were significant compared with the mice of the I/R group (all P < 0.05). The inflammatory cell recruitment was decreased in the I/R RF group. Moreover, compared with the mice of the I/RRF group, the mice of the I/RRF LY294002 group had increased levels of BUN and SCr in venous blood, decreased p-eNOS/eNOS ratio in renal tissues, increased IL-1β and IL-6 release, and increased Caspase-3 mRNA expression; and the differences were significant (all P<0.05). The inflammatory cell recruitment was increased in the I/R + RF + LY294002 group. Conclusion: RF exerts protective effect on kidney with I/R injury by alleviating renal inflammation and cell apoptosis through activating PI3K/Akt/eNOS pathway.