Endothelin 1 regulates SOCC/TGF-βand involved in atrial fibrosis in rats with atrial fibrillation / 安徽医科大学学报

Objective @#To investigate the effect and mechanism of endothelin-1 (ET-1) on atrial fibrosis in Atrial fibrillation (AF) rats .@*Methods @# Fourteen adult male SD rats were randomly divided into normal control ( NC) group and Atrial fibrillation (AF) group . The rat model of Atrial fibrillation was established by inj ecting 0.1 ml/ 100g CaCl2 Ach mixture into the tail vein once a day for one week . The control group was inj ected with the same dose of normal saline . An electrocardiogram of normal or atrial fibrillation was recorded on the first day and the eighth day in each group , and echocardiography was used to monitor atrial size and cardiac function . The fibrosis of atrial was ob served using Masson and HE staining. The expression of endothelin-1 ( ET-1) , collagen-I ( Col-I) , transforming growth factor-β(TGF-β) and the store operated calcium channel (SOCC) protein Orai1 , stromal in teraction molecule 1 (STIM1) in atrial tissue were detected by Western blot. HL-1 cells were cultured and treated with gradient concentration of ET-1 for 24 hours . Western blot was used to ob serve changes in the expression of TGF-β, Orai1 and STIM1 proteins in ET-1 /SOCC/TGF βsignaling pathway of HL-1 cells . Small interfering RNA ( siRNA) transfection method was used to knock down the expression of Orai1 in HL-1 cells , then the cells were treated with appropriate concentrations of ET-1 for 24 hours , and the expression of TGF-β protein in HL-1 cells was detected by Western blot.@*Results @#Compared with the control group , echocardiography showed a significant in crease in left atrial diameter (LAD) of the heart in atrial fibrillation rats (P < 0.05) . The HE and Masson staining results showed significant fibrosis in the myocardial tissue of AF group rats (P < 0.05) , and the Western blot re sults indicated the expression of ET-1 , Orai1 , STIM1 , TGF-β and COL-Ⅰ in the myocardial tissue of AF group significantly increased compared to the NC group (P < 0.05) . After ET 1 treatment of HL-1 cells , the protein ex pression of Orai1 , STIM1 and TGF βincreased (P < 0.05) , while knocking down Orai1 in HL-1 cells , ET-1 treat ment no longer caused the expression of TGF-β a significant upregulation .@*Conclusion @#AF caused by atrial fibril lation results in a significant increase in ET-1 expression in atrial tissue , and ET-1 /SOCC/TGF-β signal pathway promotes atrial fibrillation and fibrosis .

Effects of acupuncture on pulmonary function and airway smooth muscle spasm in asthma rats / 中国针灸

OBJECTIVE@#To observe the effects of acupuncture at "Kongzui" (LU 6) and "Yuji" (LU 10) on the latent period of inducing asthma, pulmonary function and the expression of endothelin-1 (ET-1) and metallothionein-2 (MT-2) in asthma rats, and to explore the possible mechanism of acupuncture in alleviating airway smooth muscle spasm and improving the acute attack of asthma.@*METHODS@#A total of 40 male SD rats of SPF-grade were randomly divided into a normal group, a model group, a medication group and an acupuncture group, 10 rats in each group. Except for the normal group, ovalbumin sensitization method was used to establish the asthma model in the other 3 groups. Salbutamol nebulization was adopted in the medication group, while acupuncture was applied at bilateral "Kongzui" (LU 6) and "Yuji" (LU 10) in the acupuncture group. The intervention was given once a day for 14 days in the two groups. The latent period of inducing asthma and pulmonary function were observed, the levels of ET-1 and tumor necrosis factor (TNF)-α in serum and bronchoalveolar lavage fluid (BALF) were detected by ELISA method, the morphology of the airway was observed by Masson staining, the ultrastructure of the airway smooth muscle was observed by transmission electron microscopy, the mRNA and protein expression of ET-1 and MT-2 in lung tissue was detected by real-time PCR and Western blot methods.@*RESULTS@#Compared with the normal group, in the model group, the latent period of inducing asthma was shortened (P<0.01); the airway resistance (RL) was increased while the dynamic compliance (Cdyn) was decreased (P<0.01, P<0.05); the levels of ET-1 and TNF-α in serum and BALF were increased (P<0.01); collagen fibers and collagen depositions were found around the bronchi, airway smooth muscle was thickened, the cell damage was severe and mitochondria were swollen; the mRNA and protein expression of ET-1 was increased while the mRNA and protein expression of MT-2 was decreased (P<0.01). Compared with the model group, in the acupuncture group, the latent period of inducing asthma was prolonged (P<0.05), the RL was decreased while the Cdyn was increased (P<0.01, P<0.05). Compared with the model group, in the medication group and the acupuncture group, the levels of ET-1 and TNF-α in serum and BALF were decreased (P<0.01, P<0.05); collagen fibers and collagen depositions around the bronchi were reduced, the thickened airway smooth muscle was lightened, the cell damage was improved; the mRNA and protein expression of ET-1 was decreased while the mRNA and protein expression of MT-2 was increased (P<0.01). Compared with the medication group, the mRNA expression of MT-2 was increased in the acupuncture group (P<0.05).@*CONCLUSION@#Acupuncture at "Kongzui" (LU 6) and "Yuji" (LU 10) can improve the pulmonary function and alleviate the airway smooth muscle spasm in rats with asthma. Its mechanism may be related to the down-regulation of ET-1 expression and up-regulation of MT-2 expression.

Analysis of changes in plasma endothelin-1 concentrations in patients with acute respiratory distress syndrome / 中华麻醉学杂志

Objective:To analyze the changes in plasma endothelin-1 (ET-1) concentrations in the patients with acute respiratory distress syndrome (ARDS).Methods:Fourteen patients with ARDS induced by trauma, 8 males and 6 females, aged 19-80 yr, were studied. The severity of ARDS was graded according to the Berlin definition of ARDS after admission to intensive care unit (ICU). Venous blood samples were obtained on 1st, 3rd and 5th days after admission to ICU, the plasma ET-1 concentrations were measured by radioimmunoassay, the pulmonary vascular permeability index (PVPI) was determined by PiCCO technique, and multiple organ dysfunction (MOD) score and lung injury score (LIS) were assessed. Spearman correlation of plasma ET-1 concentrations with MOD score, LIS and PVPI was analyzed.Results:MOD score, LIS, PVPI and plasma ET-1 concentrations were significantly decreased in mild ARDS patients ( n=5) as compared with moderate ARDS patients ( n=9, P<0.05). The plasma ET-1 concentration was positively correlated with MOD score, LIS and PVPI ( r=0.69, 0.76, 0.62, P<0.001). Conclusions:Plasma ET-1 concentrations can reflect the pulmonary vascular permeability and even the severity of the disease in the early stage of ARDS, so it is necessary to carry out dynamic monitoring in the patients.

Gender differences in genotypic distribution of endothelin-1 gene and endothelin receptor A gene in pulmonary hypertension associated with rheumatic mitral valve disease

Introduction: The female gender is a risk factor for idiopathic pulmonary arterial hypertension. However, it is unknown whether females with rheumatic mitral valve disease are more predisposed to develop pulmonary hypertension compared to males. Aim: We aimed to investigate whether there was a difference in genotypic distribution of endothelin-1 (ET-1) and endothelin receptor A (ETA) genes between female and male patients of pulmonary hypertension associated with rheumatic mitral valve disease (PH-MVD). Methods: We compared prevalence of ET-1 gene (Lys198Asn) and ETA gene (His323His) polymorphisms according to gender in 123 PH-MVD subjects and 123 healthy controls. Results: The presence of mutant Asn/Asn and either mutant Asn/Asn or heterozygous Lys/Asn genotypes of Lys198Asn polymorphism when compared to Lys/Lys in females showed significant association with higher risk (odds ratio [OR] 4.5; p ¼0.007 and OR 2.39; p ¼0.02, respectively). The presence of heterozygous C/T and either mutant T/T or heterozygous C/T genotypes of His323His polymorphism when compared to wild C/C genotype in females showed a significant association with higher risk (OR 1.96; p ¼0.047 and OR 2.26; p ¼0.01, respectively). No significant difference was seen in genotypic frequencies in males between PH-MVD subjects and controls. Logistic regression analysis showed that mutant genotype Asn/Asn (p ¼0.007) and heterozygous genotype Lys/Asn of Lys198Asn polymorphism (p ¼0.018) were independent predictors of development of PH in females.

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.

Upregulation of ppET-1/ETBR/eNOS mRNA Expression After Calcitriol Treatment in Chronic Kidney Diseases Model in Rats

@#Introduction: Myofibroblast formation in the interstitial area is the hallmark of chronic kidney disease (CKD). Endothelin signalling has been known to play role in physiology and pathophysiology in the kidney. Vitamin D has a reno-protective effect through inhibiting inflammation and fibrosis. However, the interaction between vitamin D and endothelin signalling in the CKD model has not been elucidated yet. Therefore, we aimed to check the difference impact of endothelin (ET) receptor in CKD. Methods: Sprague Dawley rats (3-months-old, 150-250grams) underwent 5/6 subtotal nephrectomy (SN) to induce CKD. Then, it was divided into 4 groups (each contains 6 rats): sham operation (SO), 5/6 subtotal nephrectomy (SN), calcitriol groups (0.01µg/100grBW/day (SN-D1), and 0.05µg/100grBW/day (SN-D2). Calcitriol was administered for 14 days after the surgery. The Sham Operation (SO) group was injected with NaCl. At the specified date, the rats were sacrificed and the kidneys were harvested. Fibrosis was quantified based on Sirius Red staining. Immunostaining was done for localizing fibroblast (PDGFRβ). The mRNA expressions of prepro-ET-1, endothelin receptor A (ETAR), endothelin receptor B (ETBR), and endothelial nitrite oxide synthase (eNOS) were quantified using reverse-transcriptase PCR (RT-PCR). Results: The CKD promotes an elevation of prepro-ET-1, ETBR, and eNOS, and reduction of ETAR (p<0.05) mRNA expression compared to the SO group. Administration of calcitriol (SN-D1 and SN-D2) showed the vice versa effects. However, only SN-D2 group consistently showed statistically significant differences whenever compared to either SO or SN groups. Conclusion: Calcitriol might attenuate interstitial fibrosis in CKD model via ET-1/eNOS signalling.

Inhibition of activated K-opioid receptor to myocardical hypertrophy induced by ET-1 in neonatal rats by extracellular signal-regulated kinase pathway / 中国药理学通报

Aim To investigate the inhibitory effect of kappa-opioid receptor(κ-OR)stimulation on extracellular signal-regulated kinase pathway on ET-1-induced cardiomyocyte hypertrophy in vitro cultured myocardial cells from neonatal rats.Methods Myocardial cells of neonatal rats were cultured in vitro.The hypertrophic myocytes were induced by ET-1(10 nmol·L-1)before κ-OR agonist U50488H(1 μmol·L-1)was administered.The antihypertrophic effect of κ-OR stimulation was observed in the presence of U0126(1μmol·L-1), Ro-31-8220(50 nmol·L-1)and PTX(5 mg·L-1).The cardiomyocytes volume was measured by computer photographalysis system.The relative expression of ERK1/2 was determined by Western blot.The morphological changes in cardiomyocytes were observed under an inverted phase contrast microscope.The expression of mRNA of atrial natriuretic peptide(ANP)was determined by RT-PCR.Results Compared with normal control group, ET-1 could induced cardiomyocyte hypertrophy.Compared with ET-1 model group, U50488H(1 μmol·L-1)could obviously inhibit ET-1-induced increase of the cardiomyocytes volume, expression of ANPmRNA and expression of ERK1/2, which was similar to U0126(1 μmol·L-1)and Ro-31-8220(50 nmol·L-1); however, the inhibitory effects of U50488H were partly lost when preincubated with U0126(1 μmol·L-1)and Ro-31-8220(50 nmol·L-1); the inhibitory effects of U50488H, U0126(1 μmol·L-1)and Ro-31-8220(50 nmol·L-1)were lost when preincubated with NOR-BNI.Conclusion The stimulation of kappa-opiod can inhibit myocardial hypertrophy induced by ET-1, which is possibly via attenuating ERK1/2.

Association between liver fibrosis progression and endothelin-1/nitric oxide in patients with nonalcoholic fatty liver disease / 临床肝胆病杂志

Objective To investigate whether the progression of liver fibrosis affects endothelial function in patients with nonalcoholic fatty liver disease (NAFLD), and to early identify the warning of cardiovascular diseases caused by endothelial dysfunction by liver fibrosis progression. Methods A total of 280 patients who attended the outpatient service or were hospitalized in Department of Liver Disease, Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine, from April 2019 to October 2020 were enrolled, and they were diagnosed with fatty liver disease by ultrasound and met the diagnostic criteria for NAFLD. General information and related serological markers were collected and recorded. FibroTouch technique was performed for the NAFLD patients diagnosed by ultrasound to record their fat attenuation parameter (FAP) and liver stiffness measurement (LSM), and according to LSM, the patients were divided into non-progressive fibrosis group (239 patients with LSM 0.05). Further analysis of the correlation of ET-1 and NO with each index showed that ET-1 was not correlated with age, NO, ALT, AST, GGT, total cholesterol, TG, HDL-C, low-density lipoprotein cholesterol (LDL-C), FAP, and BMI ( r s =-0.017, 0.054, -0.067, -0.016, -0.031, 0.004, 0.051, -0.084, -0.030, 0.080, and 0.044, all P > 0.05), and NO was not correlated with age, ET-1, ALT, AST, GGT, total cholesterol, TG, HDL-C, LDL-C, FAP, and BMI ( r s =0.004, 0.054, 0.011, 0.052, 0.004, -0.051, -0.052, -0.012, -0.076, -0.013, and -0.021, all P > 0.05). Conclusion This study shows that liver fibrosis progression in NAFLD has no impact on ET-1 and NO, suggesting that fibrosis progression may have no influence on endothelial function.

Endothelin-1 gene Lys198Asn polymorphism and ischemic stroke / 国际脑血管病杂志

Endothelin (ET) is a potent vasoconstrictor peptide produced by endothelial cells, which is closely associated with vascular endothelial dysfunction and cardio-cerebrovascular diseases. Recent studies have shown that ET-1 gene Lys198Asn polymorphism can be used as a new biomarker of cerebrovascular diseases. This article reviews the research progress on the relationship between the gene polymorphism and susceptibility to ischemic stroke, and discusses its clinical significance.

Serum endothelin-1 level in children with osteosarcoma and its correlation with lung metastasis / 中华实用儿科临床杂志

Objective:To investigate the serum levels, diagnosis and prognosis value of endothelin-1(ET-1) in children suffering from lung metastasis of osteosarcoma.Methods:A total of 84 children with osteosarcoma, 67 children with lung metastasis of osteosarcoma and 35 healthy people from January 1, 2013 to January 1, 2018 in Second People′s Hospital of Nanyang were retrospectively included.The serum level of ET-1 was measured by performing enzyme linked immunosorbent assay(ELISA) methods and the influencing factors of serum ET-1 levels in children with lung metastasis of osteosarcoma were conducted by Logistic regression analysis.The clinical value of ET-1 in the prediction of the incidence of lung metastasis in children with osteosarcoma was analyzed by receiver operating characteristic (ROC) curve.Forty-five children with lung metastasis of osteosarcoma were followed up for 18 months and the prognosis value of serum ET-1 levels in children with lung metastasis of osteosarcoma was evaluated by Kaplan-Meier survival analysis. Results:The serum ET-1 level in children with lung metastasis of osteosarcoma was 97.23 (65.13, 134.98) ng/L and significantly higher than osteosarcoma group 60.21 (43.12, 74.63) ng/L and healthy control group 34.45 (12.01, 63.03) ng/L, respectively ( Z=-5.671, -4.92, all P<0.05), with significant differences. Logistic regression analysis proved that lung bilateral involvement ( OR=3.449), numbers of lung metastases (more than 3)( OR=3.449), average diameter of lung metastases (more than 5 cm) ( OR=6.501) and extrapulmonary metastasis ( OR=4.369) were independent risk factors for elevated serum ET-1 levels in children developing lung metastasis of osteosarcoma.The predictive value of ET-1 in the incidence of lung metastasis in children with osteosarcoma was significant (area under ROC curve: 0.841). When the cut-off value was 94.27 ng/L, the sensitivity and specificity were 88.5% and 92.6%, respectively.Survival analysis revealed that higher levels of ET-1 was correlated with poor prognosis ( OR=3.287, 95% CI: 1.119-9.547). Conclusions:The serum levels of ET-1 in children with lung metastasis of osteosarcoma are significantly elevated.ET-1 is a serological marker for the differential diagnosis of lung metastasis of osteosarcoma.Moreover, the higher levels of ET-1 are correlated with poor prognosis in children with lung metastasis of osteosarcoma.

Comparison of intravenous anesthesia between propofol and etomidate in patients undergoing laparoscopic surgery and their effects on plasma nitric oxide and endothelin-1 / 中国医师进修杂志

Objective:To compare the effects of intravenous anesthesia between propofol and etomidate in patients undergoing laparoscopic surgery and their effects on plasma nitric oxide (NO) and endothelin-1 (ET-1).Methods:The clinical data of 80 patients with laparoscopic surgery in Guangrao People′s Hospital from March 2017 to March 2019 were retrospectively analyzed. Among them, 40 cases were given propofol intravenous anesthesia (propofol group), and 40 cases were given etomidate intravenous anesthesia (etomidate group). The anesthetic effect, plasma NO and ET-1 levels, hemodynamic indexes and adverse reactions (muscle spasm, nausea and vomiting, injection site pain, body movement and respiratory depression) were compared between the two groups.Results:The time of consciousness disappearance, tracheal intubation, eye opening, spontaneous breathing and speech response in etomidate group were significantly shorter than those in propofol group: (57.48 ± 2.63) s vs. (86.17 ± 7.41) s, (4.39 ± 2.56) min vs. (6.42 ± 2.58) min, (5.39 ± 2.56) min vs. (9.42 ± 2.58) min, (5.21 ± 1.99) min vs. (8.75 ± 2.54) min and (8.39 ± 2.56) min vs. (8.39 ± 2.56) min, and the differences were statistically significant ( P<0.05). The levels of NO and ET-1 in the etomidate group were significantly lower than those in the propofol group at 0.5, 1.0 and 1.5 h after pneumoperitoneum ( P< 0.05). The levels of systolic blood pressure, diastolic blood pressure and oxygen saturation (SpO 2) in the etomidate group were significantly higher than those in the propofol group: (78.42 ± 4.68) mmHg (1 mmHg = 0.133 kPa) vs. (74.11 ± 6.63) mmHg, (132.86 ± 8.71) mmHg vs. (111.24 ± 3.56) mmHg and 0.982 ± 0.032 vs. 0.953 ± 0.043, and the differences were statistically significant ( P<0.05). The incidence of adverse reactions in the etomidate group was significantly lower than that in the propofol group: 17.5% (7/40) vs. 47.5% (19/40), P<0.05. Conclusions:Compared with propofol intravenous anesthesia, etomidate intravenous anesthesia in laparoscopic surgery patients has more stable hemodynamics and better anesthetic effect. It can effectively inhibit the release of NO and ET-1, and has higher safety.

Efficacy of triamcinolone acetonide injection behind the eyeball in the treatment of Purtscher disease and its effect on vision, inflammatory factors, nitric oxide and endothelin-1 of aqueous fluid / 中国医师进修杂志

Objective:To investigate the efficacy of triamcinolone acetonide injection behind the eyeball in the treatment of Purtscher disease and its effects on vision, inflammatory factors, nitric oxide (NO) and endothelin-1 (ET-1) of aqueous fluid.Methods:A total of 64 patients with Purtscher disease admitted to First People′s Hospital of Lanzhou City from January 2016 to January 2020 were selected and divided into two groups by random number table, with 32 cases in each group. The control group was given conventional treatment, and the observation group was injected with triamcinolone acetonide behind the eyeball on this basis. The total effective rate after 6 weeks of treatment, the best corrected visual acuity and hemodynamic indicators peak flow velocity (V max), end-diastolic flow velocity (V min), resistance index (RI) of ophthalmic artery and central retinal artery during systole, aqueous humor inflammatory factors tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-2, IL-6, NO, ET-1, and adverse reactions after 3 and 6 weeks of treatment were compared between the two groups. Results:After 6 weeks of treatment, the total effective rate in the observation group was higher than that in the control group: 96.9% (31/32) vs. 75.0% (24/32), and the difference was statistically significant ( χ2 = 4.655, P<0.05). After 3, 6 weeks of treatment, the best corrected visual acuity in the observation group was higher than that in the control group: 0.44 ± 0.13 vs. 0.35 ± 0.12, 0.88 ± 0.16 vs. 0.62 ± 0.15, and the difference was statistically significant ( P<0.05). After 3, 6 weeks of treatment, the V max and V min of the ophthalmic artery and central retinal artery in the observation group were higher than those in the control group: after 3 weeks of treatment: (29.42 ± 0.93) cm/s vs. (28.65 ± 0.90) cm/s, (8.04 ± 1.08) cm/s vs. (6.95 ± 1.12) cm/s, (8.42 ± 1.21) cm/s vs. (8.39 ± 1.19) cm/s, (4.05 ± 0.61) cm/s vs.(3.58 ± 0.57) cm/s; after 6 weeks of treatment: (31.72 ± 1.03) cm/s vs. (29.58 ± 0.95) cm/s, (9.81 ± 1.06) cm/s vs. (7.78 ± 1.15) cm/s, (12.01 ± 1.23) cm/s vs. (10.82 ± 1.07) cm/s, (4.65 ± 0.47) cm/s vs. (4.02 ± 0.36) cm/s; the RI of the ophthalmic artery and central retinal artery in the observation group were lower than those in the control group: after 3 weeks of treatment: 0.76 ± 0.06 vs. 0.80 ± 0.05, 0.70 ± 0.03 vs. 0.76 ± 0.04; after 6 weeks of treatment: 0.71 ± 0.05 vs.0.75 ± 0.04, 0.68 ± 0.04 vs.0.72 ± 0.03, and the differences were statistically significant ( P<0.05). After 3 and 6 weeks of treatment, the levels of TNF-α, IL-1β, IL-2, IL-6 of aqueous fluid in the observation group werelower than those in the control group: after 3 weeks of treatment: (2.61 ± 0.38) μg/L vs.(4.75 ± 0.41) μg/L, (3.45 ± 0.28) μg/L vs.(6.28 ± 1.05) μg/L, (6.28 ± 0.34) μg/L vs. (9.45 ± 1.14)μg/L, (9.82 ± 1.66) ng/L vs. (12.45 ± 2.31) ng/L; after 6 weeks of treatment: (0.98 ± 0.23) μg/L vs. (1.44 ± 0.27) μg/L, (1.11 ± 0.36) μg/L vs. (1.69 ± 0.38) μg/L, (3.68 ± 1.02) μg/L vs. (5.73 ± 1.15) μg/L, (4.11 ± 1.19) ng/L vs. (7.38 ± 1.06) ng/L, and the differences were statistically significant ( P<0.05). After 3, 6 weeks of treatment, the levels of NO in the observation group washigher than that in the control group: (56.49 ± 9.17) μmol/L vs. (50.22 ± 8.68) μmol/L, (60.18 ± 8.27) μmol/L vs. (53.24 ± 7.96) μmol/L; the levels of ET-1 waslower than that in the control group: (60.77 ± 9.61) ng/L vs. (65.02 ± 10.85) ng/L, (56.59 ± 7.22) ng/L vs. (61.18 ± 6.76) ng/L, and the differences were statistically significant ( P<0.05). The incidence of adverse reactions in the two groups had no statistical significance ( P>0.05). Conclusions:The injection of triamcinolone acetonide behind the eyeball to treat Purtscher disease can improve ocular hemodynamics, inhibit inflammation, improve vascular endothelial function and improve visual acuity. It has significant curative effect and certain safety.

Effect of electroacupuncture on choroidal blood flow and the expression of endothelin-1 and its receptor in lens-induced myopic guinea pigs / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To observe the changes of the vessel density of choriocapillaris in lens-induced myopia in guinea pigs, and to explore choroidal endothelin-1(ET-1), endothelin receptor A(ETAR)and receptor B(ETBR)expression changes and the effect of electroacupuncture. <p>METHODS: Fifty-four guinea pigs were randomly divided into normal control(NC), lens-induced myopia(LIM)and LIM+electroacupuncture(LIM+EA). The NC group was fed normally without intervention and the right eye in LIM group and LIM+EA group was coverd with a -6.00D lens to establish a myopia model. At 2 and 4wk, the refraction, axial length and the vessel density of choriocapillaris in groups were measured. The expression and protein content of ET-1, ETAR and ETBR mRNA in groups were detected by the real-time fluorescent quantitative PCR(quantitative polymerase chain reaction, q-PCR), enzyme-linked immunosorbent assay(ELISA)and immunohistochemistry. <p>RESULTS: At 2 and 4wk, compared with the the NC group, refraction and axial length in LIM group and LIM+EA group had significantly increased(all <i>P</i><0.001). Compared with the LIM group, the refraction and axial length in LIM+EA group were decreased(all <i>P</i><0.05). At 2 and 4wk, compared with the NC group,the vessel density of choriocapillaris was decreased(<i>P</i><0.001)and the ET-1, ETAR and ETBR mRNA and protein levels in choroid were increased(all <i>P</i><0.05)in LIM group. At 2 and 4wk, compared with the LIM group,the vessel density of choriocapillaris was decreased(<i>P</i><0.01)and the ET-1, ETAR and ETBR mRNA and protein levels in choroid were increased in LIM+EA group.<p>CONCLUSION:In LIM guinea pigs, the choroidal blood flow decreased with the increased of refraction and axial length, which may affect ET-1 and its receptors through vascular shear force during the development of myopia. At the same time, electroacupuncture can improve choroidal blood flow through neuromodulation and affects the vascular shear stress to down-regulate the content of ET-1 and its receptor to delay the development of myopia.

Effect of acupuncture on vascular endothelial function in patients of polycystic ovary syndrome with different glucose tolerance status / 中国针灸

OBJECTIVE@#To observe the effect of acupuncture on vascular endothelial function in patients of polycystic ovary syndrome (PCOS) with impaired glucose tolerance (IGT) and normal glucose tolerance (NGT).@*METHODS@#A total of 140 patients with PCOS were divided into an IGT group (70 cases, 11 dropped off) and a NGT group (70 cases, 9 cases dropped off). The patients in the two groups were treated with full-cycle acupuncture at Zhongwan (CV 12), Guanyuan (CV 4), Qihai (CV 6), Tianshu (ST 25), etc. once every other day, 3 times a week, for 3 months. Before and after treatment, TCM symptom score, insulin resistance index [including fasting plasma glucose (FPG), 2-hour blood glucose (2hPG), fasting serum insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR)] and vascular endothelial related factors [including asymmetric dimethylarginine (ADMD), endothelin-1 (ET-1), malondialdehyde (MDA), nitric oxide (NO)] were compared between the two groups; in addition, the obese subgroup and non-obese subgroup of the two groups were further compared.@*RESULTS@#Compared before treatment, the TCM symptom scores, ADMD, ET-1 and MDA after treatment were decreased (@*CONCLUSION@#Acupuncture could improve vascular endothelial function in PCOS patients, IGT patients have better efficacy than NGT patients, and obese patients have better efficacy than non-obese patients.

Relationship and risk factors of osteonecrosis of femoral head after internal fixation of femoral neck fracture with serum bone metabolism and vasoactive factors / 中国骨伤

OBJECTIVE@#To study the relationship between osteonecrosis of femoral head after internal fixation of femoral neck fracture and serum bone metabolism, vascular active factors, and analyze the risk factors.@*METHODS@#Total 150 patients with femoral neck fracture who underwent reduction and internal fixation from April 2016 to April 2019 were selected, including 83 males and 67 females. According to whether there was necrosis of femoral head after operation, they were divided into necrosis group(32 cases) and non necrosis group (118 cases). Before operation and 1, 3, 5 days after operation, the serum levels of beta-C terminal cross-linked telopeptides of typeⅠ collagen(β-CTX), N-telopeptide of typeⅠ procollagen(PINP), nitric oxide (NO), Endothelin-1 (ET-1) were measured. The clinical characteristics of the two groups were compared. The risk factors of postoperative femoral head necrosis were analyzed by logistic regression model. The value of serum indexes in predicting postoperative femoral head necrosis was analyzed by ROC curve.@*RESULTS@#There was no significant difference in the levels of serum PINP and β-CTX between necrotic group and non necrotic group before operation and 1, 3 and 5 days after operation(@*CONCLUSION@#The content of serum NO and ET-1 on the first day after operation can predict the necrosis of femoral head.

Role of FAK signaling in chagasic cardiac hypertrophy

Abstract Cardiac hypertrophy and dysfunction are a significant complication of chronic Chagas disease, with heart failure, stroke, and sudden death related to disease progression. Thus, understanding the signaling pathways involved in the chagasic cardiac hypertrophy may provide potential targets for pharmacological therapy. Herein, we investigated the implication of focal adhesion kinase (FAK) signaling pathway in triggering hypertrophic phenotype during acute and chronic T. cruzi infection. C57BL/6 mice infected with T. cruzi (Brazil strain) were evaluated for electrocardiographic (ECG) changes, plasma levels of endothelin-1 (ET-1) and activation of signaling pathways involved in cardiac hypertrophy, including FAK and ERK1/2, as well as expression of hypertrophy marker and components of the extracellular matrix in the different stages of T. cruzi infection (60-210 dpi). Heart dysfunction, evidenced by prolonged PR interval and decrease in heart rates in ECG tracing, was associated with high plasma ET-1 level, extracellular matrix remodeling and FAK signaling activation. Upregulation of both FAK tyrosine 397 (FAK-Y397) and serine 910 (FAK-S910) residues phosphorylation as well as ERK1/2 activation, lead to an enhancement of atrial natriuretic peptide gene expression in chronic infection. Our findings highlight FAK-ERK1/2 signaling as a regulator of cardiac hypertrophy in Trypanosoma cruzi infection. Both mechanical stress, induced by cardiac extracellular matrix (ECM) augment and cardiac overload, and ET-1 stimuli orchestrated FAK signaling activation with subsequent activation of the fetal cardiac gene program in the chronic phase of infection, highlighting FAK as an attractive target for Chagas disease therapy.

Imbalance between Endothelin-1 and eNOS Expression Associates with Tubular Injury in Mice with 5/6 Subtotal Nephrectomy

@#Introduction: Chronic Kidney Diseases (CKD) leads to kidney fibrosis which characterized by tubular injury and atrophy with interstitial fibrosis. Endothelin-1 (ET-1) and endothelial Nitrite Oxide Synthase (eNOS) are known to play role in CKD and kidney fibrosis, although their correlation with tubulo-interstitial injury have not been understood clearly. Methods: 5/6 Subtotal Nephrectomy (SN) was performed in male Swiss Background mice to induce CKD. Sham operation (SO, n=5) procedure was performed on mice as control. The mice were sacrificed in day 7 (1N, n=5) and day 28 (4N, n=5) after operation. We measured creatinine serum to assess renal function. Tubular injury score was quantified based on Periodic-Acid Schiff (PAS) staining. Prepro-ET-1 and eNOS were quantified using RT-PCR. Results: SN_1N and SN_4N groups had significant higher of serum creatinine and tubular injury from SO group. Densitometry analysis of RT-PCR revealed up-regulation of prepro-ET-1 mRNA expression in SN_1N and SN_4N (p<0.05 vs SO). Meanwhile, we found a significant increase of eNOS expression in SN_1N, and then it reduced significantly in SN_4N. We found significant parallel correlation between ET-1 and tubular injury expression (r: 0.768;p<0,05), meanwhile there were insignificant inverse correlation between eNOS and tubular injury (r: -0.354;p>0.05). Conclusion: eNOS might play role as a counterbalance in the up regulation of ET-1 in acute condition after SN. However, it failed in chronic condition. These lead to deterioration of renal function and tubular injury. An imbalance between ET-1 and eNOS expression in chronic CKD model might play role in profound renal damage.

Antioxidant effect of endothelin-1 receptor antagonist protects the rat kidney against chronic injury induced by hypertension and hyperglycemia / Efeito antioxidante de antagonista dos receptores de endotelina-1 protege ratos contra lesão renal crônica induzida por hipertensão e hiperglicemia

ABSTRACT Hypertension and Diabetes mellitus are the two main causes of chronic kidney disease that culminate in the final stage of kidney disease. Since these two risk factors are common and can overlap, new approaches to prevent or treat them are needed. Macitentan (MAC) is a new non-selective antagonist of the endothelin-1 (ET-1) receptor. This study aimed to evaluate the effect of chronic blockade of ET-1 receptor with MAC on the alteration of renal function observed in hypertensive and hyperglycemic animals. Genetically hypertensive rats were divided into control hypertensive (HT-CTL) group, hypertensive and hyperglycemic (HT+DIAB) group, and hypertensive and hyperglycemic group that received 25 mg/kg macitentan (HT-DIAB+MAC25) via gavage for 60 days. Kidney function and parameters associated with oxidative and nitrosative stress were evaluated. Immunohistochemistry for neutrophil gelatinase-associated lipocalin (NGAL), ET-1, and catalase in the renal cortex was performed. The HT+DIAB group showed a decrease in kidney function and an increase in NGAL expression in the renal cortex, as well as an increase in oxidative stress. MAC treatment was associated with attenuated ET-1 and NGAL production and increases in antioxidant defense (catalase expression) and nitric oxide production. In addition, MAC prevented an increase in oxidant injury (as measured by urinary hydroperoxide and lipid peroxidation), thus improving renal function. Our results suggest that the antioxidant effect of the ET-1 receptor antagonist MAC is involved in the improvement of kidney function observed in hypertensive and hyperglycemic rats.

RESUMO Hipertensão e Diabetes Mellitus figuram como as duas principais causas de doença renal crônica que culmina em doença renal terminal. Uma vez que os dois fatores de risco são comuns e podem se sobrepor, novas abordagens preventivas e terapêuticas se fazem necessárias. O macitentan (MAC) é um novo antagonista não-seletivo dos receptores da endotelina-1 (ET-1). O presente estudo teve como objetivo avaliar os efeitos do bloqueio crônico dos receptores da ET-1 com MAC sobre a alteração da função renal em animais hipertensos e hiperglicêmicos. Ratos geneticamente hipertensos foram divididos em grupos com animais hipertensos de controle (HT-CTL), hipertensos e hiperglicêmicos (HT+DIAB) e hipertensos e hiperglicêmicos tratados com 25 mg/kg de macitentan (HT-DIAB+MAC25) via gavagem por 60 dias. Foram avaliados função renal e parâmetros associados ao estresse oxidativo e nitrosativo. Exames de imunoistoquímica foram realizados para lipocalina associada à gelatinase neutrofílica (NGAL), ET-1 e catalase no córtex renal. O grupo HT+DIAB exibiu diminuição da função renal e aumento na expressão de NGAL no córtex renal, bem como estresse oxidativo aumentado. O tratamento com MAC foi associado a atenuação da produção de ET-1 e NGAL e maior ativação das defesas antioxidantes (expressão de catalase) e elevação da produção de óxido nítrico. Além disso, o MAC evitou exacerbação da lesão oxidante (medida por hidroperóxidos urinários e peroxidação lipídica), melhorando assim a função renal. Nossos resultados sugerem que o efeito antioxidante do antagonista dos receptores da ET-1 MAC esteja imbricado no aprimoramento da função renal observada em ratos hipertensos e hiperglicêmicos.

Effect of Danggui Shaoyao San Drug-containing Serum on Expression of p-MLCⅡ and MLCⅡ Protein in HSC-T6 Cells Induced by ET-1 / 中国实验方剂学杂志

Objective:To investigate the effect of endothelin-1 (ET-1) on the expression of phosphorylated myosin light chain Ⅱ(p-MLCⅡ)and myosin light chain Ⅱ(MLCⅡ)protein in rat hepatic stellate cells HSC-T6 and explore the intervention effect of Danggui Shaoyao San(DSS)drug-containing serum. Method:After HSC-T6 cells were seeded, DMEM and blank rat serum with final concentrations of 2.5%, 5%, 10%, 15% and 20% were added to each well. The viability of HSC-T6 cells was determined by methyl thiazolyl tetrazolium(MTT) assay to screen the suitable serum concentration range. The cells were divided into blank serum control group (5%, 10%, 15%) and DSS drug-containing serum group (5%, 10%, 15%). ELISA was used to detect the content of ET-1 in cell culture supernatant under basic state. The cells were divided into blank serum control group (10%), DSS drug-containing serum low (5%), medium (10%) and high dose (15%) groups. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) was used to detect the level of ET-1 mRNA in cell culture supernatant under basic state. The cells were divided into blank serum control group (10%), model group (10%), DSS drug-containing serum low (5%), medium (10%), high dose (15%) groups and Y-27632 inhibitor group (100 μmol·L-1). Except the blank serum control group, the other groups all received 10 nmol·L-1 ET-1 to induce HSC-T6 cells. Western blot was used to detect the expression of p-MLCⅡ and MLCⅡ in HSC-T6 cells induced by ET-1. Result:Serum concentrations of 5%, 10% and 15% were used as drug-containing serum concentrations. As compared with the blank serum control group, the DSS drug-containing serum group significantly reduced the relative content of ET-1 and ET-1 mRNA in the basic state (PPPPPConclusion:DSS drug-containing serum may down-regulate the expression of p-MLCⅡ and MLCⅡ by down-regulating the content of ET-1 and inhibiting the autocrine of ET-1.