Effects of early-life exposure to angiotensinⅡ type 1 receptor autoantibody on lipid metabolism in offspring rats / 中华围产医学杂志

Objective:To investigate the effects of early-life (intrauterine and breastfeeding period) exposure to angiotensin Ⅱ type 1 receptor autoantibody (AT 1-AA) on lipid metabolism in offspring rats. Methods:Thirty-two AT 1-AA negative healthy nonpregnant specific pathogen free female Sprague Dawley rats weighing 150-170 g were randomly divided into two groups. Those in the immune group ( n=16) were subcutaneously injected with the mixture of an equal volume of Freund's adjuvant and the second extracellular loop of human-derived angiotensin Ⅱ receptor type 1 (AT1R-ECⅡ) repeatedly to establish the AT 1-AA-positive rat model by active immunization and those in the control group ( n=16) with normal saline solution. Before each immunization, blood samples were collected from the tail of rats to detect serum AT 1-AA levels of those rats in both groups, and the AT 1-AA-positive rat model was successfully established when the serum AT 1-AA was positive and its level reached a plateau. After eight weeks of immunization, the female rats in the two groups were mated with healthy AT 1-AA-negative male rats to conceive. Serum samples were collected from the maternal and offspring rats at the gestation of 18 days (G18), postnatal 21 days (P21), and from the normally fed offspring rats from the time of weaning to 12 weeks old (W12). Active immunization was not performed on the offspring throughout the experiment. The serum AT 1-AA levels of maternal and offspring rats were determined by enzyme-linked immunosorbent assay, and serum AT1-AA was positive when the ratio of AT1-AA level of the immune group over the control group ≥2.1. The blood lipid levels of maternal and offspring rats were measured by an automatic biochemical analyzer. Serum AT 1-AA levels, total cholesterol (TC), high-density lipoprotein-cholesterol [instead of high-density lipoprotein (HDL)], low-density lipoprotein-cholesterol, and free fatty acid levels of the offspring and maternal rats were determined for correlation analysis. Two independent sample t-test, linear regression analysis, and analysis of variance were adopted for statistical analysis. Results:(1) The serum levels of AT 1-AA in maternal rats at G18 and P21 in the immune group were significantly higher than those in the control group (G18: 1.170±0.190 vs 0.114±0.016, t=14.64; P21: 0.988±0.283 vs 0.084±0.006, t=9.57; both P<0.001). (2) The serum levels of AT 1-AA in the offspring at G18 and P21 in the immune group were significantly higher than those in the control group (offspring at G18: 0.948±0.220 vs 0.105±0.010, t=10.10; male offspring at P21: 0.758±0.273 vs 0.080±0.002, t=7.46; female offspring at P21: 0.774±0.274 vs 0.084±0.005, t=7.55; all P<0.001), which showed a positive correlation with those in maternal rats at the same period (offspring at G18: R=0.78; male offspring at P21: R=0.82; female offspring at P21: R=0.82; all P<0.05). However, there was no significant difference in the serum AT 1-AA level in offspring at W12 between the immune and control group ( P>0.05). (3) The serum levels of TC at G18 and P21, and HDL at P21 in maternal rats in the immune group were all higher than those in the control group [TC at G18: (2.36±0.32) vs (1.95±0.24) mmol/L, t=2.70; P21: (2.82±0.50) vs (2.18±0.26) mmol/L, t=3.41; HDL at P21: (1.94±0.33) vs (1.57±0.23) mmol/L, t=2.80; all P<0.05]. (4) Compared with the offspring in the control group, there was no significant change in lipid metabolism at G18 and W12 in the offspring in the immune group (both P>0.05). The serum levels of TC and HDL in male and female offspring at P21 in the immune group were higher than their counterparts in the control[TC in male offspring: (2.38±0.52) vs (1.83±0.30) mmol/L, t=2.73; HDL in male offspring: (1.44±0.32) vs (1.07±0.18) mmol/L, t=2.98; TC in female offspring: (2.50±0.72) vs (1.70±0.26) mmol/L, t=3.16; HDL in female offspring: (1.41±0.33) vs (1.00±0.14) mmol/L, t=3.41; all P<0.05]. (5) The serum levels of TC and HDL in male and female offspring at P21 in the immune group showed no correlation with those in maternal rats at P21 (all R<0.5, all P>0.05). The serum levels of HDL in male and female offspring at P21 in the immune group had a positive correlation with their own serum TC levels (male offspring: R=0.98; female offspring: R=0.97; both P<0.001) and also with their own serum AT 1-AA levels (male offspring: R=0.74, P=0.023; female offspring: R=0.91, P=0.001). The serum levels of TC in male and female offspring at P21 in the immune group had a positive correlation with their serum AT 1-AA levels (male offspring: R=0.72, P=0.030; female offspring: R=0.90, P=0.001). Conclusion:The early-life exposure to AT 1-AA may cause abnormal expression of TC and HDL in offspring rats.

Relationship between edaravone-induced inhibition of pressure overload-induced myocardial remodeling and AT1R/MAPKs/StAR signaling pathway in rats / 中华麻醉学杂志

Objective:To evaluate the relationship between edaravone-induced inhibition of pressure overload-induced myocardial remodeling and angiotensin Ⅱ type 1 receptor (AT1R)/mitogen activated protein kinases (MAPKs)/steroidogenic acute regulatory protein (StAR) signaling pathway in rats.Methods:Thirty-six clean-grade healthy male Sprague-Dawley rats, aged 2 months, weighing 200-220 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group (S group), pressure overload group (POL group) and edaravone group (E group). The cardiac pressure overload was induced by ligation of thoracic aorta for 8 weeks.After the model preparation, 0.9% sodium chloride 10 ml/kg was intraperitoneally injected daily in group POL, and edaravone 10 mg/kg was given instead in group E for 8 consecutive weeks.After the model was successfully established, the left ventricular ejection fraction (EF) and ventricular shortening fraction (FS) were measured by two-dimensional ultrasound.The animals were sacrificed by bloodletting, and the heart weight/body weight ratio (HW/BW ratio) was calculated.Myocardial tissues were obtained for determination of the cross-sectional area (MSA) after HE staining, the collagen volume fraction (CVF) (using Masson′s staining), the expression of AT1R and StAR (by immunohistochemistry), extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 MAPK phosphorylation levels (p-ERK1/2/ERK1/2 ratio and p-p38 MAPK/p38 MAPK ratio) (by Western blot) and the aldosterone content (by enzyme-linked immunosorbent assay). Results:Compared with group S, the HW/BW ratio, MSA and CVF were significantly increased, EF and FS were decreased, AT1R and StAR expression was up-regulated, and p-ERK1/2/ERK1/2 ratio, p-p38 MAPK/p38 MAPK ratio and aldosterone content were increased in group POL ( P<0.05). Compared with POL group, the HW/BW ratio, MSA and CVF were significantly decreased, EF and FS were increased, AT1R and StAR expression was down-regulated, and p-ERK1/2/ERK1/2 ratio, p-p38 MAPK/p38 MAPK ratio and aldosterone content were decreased in group E ( P<0.05). Conclusion:The mechanism of edaravone-induced inhibition of pressure overload-induced myocardial remodeling is probably associated with inhibiting the activation of AT1R/MAPKs/StAR signaling pathway in rats.

Impacto del COVID-19 en las enfermedades hepáticas y la salud pública en el Perú / Impact of COVID-19 on liver disease and the public health in Peru

RESUMEN La pandemia declarada por la OMS originada por el COVID-19 (enfermedad infecciosa originada por el virus SARS-CoV2), desde su aparición en Wuhan, China en diciembre 2019; esta diseminándose rápidamente y de manera inesperada por todo el mundo originando millones de casos y miles de muertes, afectando más de 120 países y desde el 06 marzo 2020 al Perú, distribuyéndose rápidamente por todo el país, originando crisis y colapso del sistema de servicios de salud, especialmente de las atenciones en emergencia, hospitalizaciones y unidades de cuidados intensivos abarrotadas; sin tener aún un tratamiento específico ni la posibilidad de una vacuna a corto plazo. Se sabe actualmente que COVID-19, es una enfermedad sistémica que puede afectar múltiples órganos y tejidos y que puede ser fatal. El objetivo de esta revisión es mostrar lo descrito en los recientes estudios publicados a nivel mundial incluido nuestro país, que han reportado sus manifestaciones clínicas, esbozando posibles mecanismos de disfunción hepática relacionados a COVID-19 y sus repercusiones, en especial sobre el aparato digestivo; analizando y discutiendo el potencial impacto sobre ellas y las enfermedades del hígado, enunciando las recomendaciones de expertos y organizaciones científicas respecto a medidas de prevención, control y manejo, además de esbozar algunas estrategias de salud pública en nuestro país para la adecuada atención de estos pacientes en tiempos de crisis generalizada.

ABSTRACT The pandemic of COVID-19 (an infectious disease caused by the SARS-CoV2 virus), declared as such by the WHO, is spreading since its appearance in Wuhan (China) in December 2019, rapidly and unexpectedly throughout the world, causing millions of cases and thousands of deaths and has affected more than 120 countries. It was officially acknowledged in Peru on March 6th, 2020, and has spread rapidly throughout the country, causing first the crisis and then the collapse of the healthcare system, especially emergency care, admissions, and overcrowded intensive care units, not having a specific treatment or the foreseeable possibility of a short-term vaccine. COVID-19 is currently known for being a systemic disease that can affect multiple organs and tissues and can be fatal. The goal of this review is to present what has been described in recent studies, published worldwide and including our country, that have reported clinical manifestations, outlining possible mechanisms of liver dysfunction related to COVID-19 and its repercussions, especially on the digestive system. These studies analyze and discuss the potential impact on liver diseases, offering recommendations of experts and scientific organizations regarding prevention, control and management measures, outlining also some public health strategies in our country for the proper care of COVID-19 patients in times of widespread crisis.

Evaluation of renin and angiotensin II levels in pseudoexfoliation syndrome / Avaliação dos níveis de renina e angiotensina II na síndrome de pseudo-exfoliação

ABSTRACT Purpose: Pseudoexfoliation syndrome has been linked to impaired function of the heart and blood vessels. We conducted a study to investigate the role of the renin-angiotensin system in the etiopathogenesis of pseudoexfoliation syndrome. Methods: The subjects were 14 patients with pseudoexfoliation syndrome and 14 healthy controls who underwent cataract extraction. Preoperative 5-ml samples of peripheral venous blood and perioperative aqueous humor were collected from the patients in both groups. Plasma and aqueous humor renin levels were analyzed by an immunoradiometric method, and angiotensin II levels were analyzed by radioimmunassay. SPSS version 16.0 was used for statistical analyses. A p-value <0.05 was considered to indicate a statistically significant difference. Results: The mean ages of the patients in pseudoexfoliation and control groups were 71.7 ± 7.1 and 67.4 ± 9.3 years, respectively (p=0.140). The median aqueous humor renin level was 7.73 pg/ml (4.15-21) in the control group and 11.95 pg/ml (3.75-18.54) in pseudoexfoliation group (p=0.022). There were no differences between the two groups in the plasma renin, plasma angiotensin II, or aqueous humor angiotensin II levels. The correlations between plasma and aqueous humor renin levels and between plasma and aqueous humor angiotensin II levels were examined separately for each group; no significant correlations were observed in pseudoexfoliation group (r=-0.440, p=0.115; r=-0.414, p=0.142) or the control group (r=-0.232, p=0.425; r=0.482, p=0.081). Conclusion: Aqueous humor renin levels are higher in pseudoexfoliation syndrome. The results indicate a probable role of renin-angiotensin system in pseudoexfoliation syndrome. Further studies with larger numbers of cases are needed to clarify the precise association of renin-angiotensin system with the etiopathogenesis of pseudoexfoliation syndrome.

RESUMO Objetivo: A síndrome de pseudo-exfoliação tem sido associada ao comprometimento da função do coração e dos vasos sanguíneos. Foi realizado um estudo para investigar o papel do sistema renina-angiotensina na etiopatogenia da síndrome de pseudo-exfoliação. Métodos: Os sujeitos foram 14 pacientes com síndrome de pseudo-exfoliação e 14 controles saudáveis submetidos à extração de catarata. Amostras pré-operatórias de 5 ml de sangue venoso periférico e humor aquoso perioperatório foram coletadas dos pacientes em ambos os grupos. Os níveis de renina no plasma e humor aquoso foram analisados pelo método imunorradiométrico e os níveis de angiotensina II foram analisados por radioimunoensaio. O SPSS versão 16.0 foi utilizado para análises estatísticas. Considerou-se o valor de p<0,05 para indicar uma diferença estatisticamente significativa. Resultados: A média de idade dos pacientes nos grupos pseudo-exfoliação e controle foi de 71,7 ± 7,1 e 67,4 ± 9,3 anos, respectivamente (p=0,140). O nível médio de renina no humor aquoso foi de 7,73 pg / ml (4,15-21) no grupo controle e 11,95 pg/ml (3,75-18,54) no grupo pseudo-exfoliação (p=0,022). Não houve diferenças entre os dois grupos de renina plasmática, angiotensina II plasmática ou nos níveis de angiotensina II em humor aquoso. As correlações entre os níveis de renina no plasma e no humor aquoso e entre os níveis de angiotensina II no plasma e humor foram examinadas separadamente para cada grupo; n]ao foram observadas correlações significativas no grupo pseudo-exfoliação (r=-0,440, p=0,115; r=-0,414, p=0,142) ou no grupo controle (r=-0,232, p=0,425; r=0,482, p=0,081). Conclusão: Os níveis de renina no humor aquoso são mais elevados na síndrome de pseudo-exfoliação. Os resultados indicam um provável papel do sistema renina-angiotensina na síndrome de pseudo-exfoliação. Novos estudos com maior número de casos são necessários para esclarecer a associação precisa do sistema renina-angiotensina com a etiopatogenia da síndrome de pseudo-exfoliação.

The expressions and clinical relevance of angiotensin-receptor-1 and hypoxia-inducible factor-1α in glomerulus and juxtaglomerular apparatus of patients with lupus nephritis / 中华风湿病学杂志

Objective To detect the expressions of angiotensin-receptor-1 (AT1R) and hypoxia-inducible factor (HIF)-1αin glomeruli and juxtaglomerular apparatus of different types of lupus nephritis (LN) patients, and analyze the correlation between them with systemic lupus erythematosus disease activity index (SLEDAI) complement 3, serum creatinine and 24-hour proteinuria in order to explore the role of the two factors in the pathogenesis of lupus nephritis (LN). Methods Between May 2010 and April 2016, a total of 90 patients with LN and 8 healthy controls were selected from Department of Rheumatology, Qujing Affiliated Hospital of Kunming Medical University and the First Affiliated Hospital of Kunming Medical University. The expressions of AT1R and HIF-1αin renal biopsy specimens were measured by streptavidin-perosidase (SP) of immunohistochemical stains. Pathological graphic analysis system was used for semi-quantitative estimate. Levels of SLEDAI, C3, serum creatinin and 24-hour proteinuria were also detected. Finally the relationshipbetween the two factors with clinical data was analyzed. The ANOVA test was used for intergroup comparison, and SNK-q test was used for the two groups comparison. Pearson's analysis was used for correlation analysis. Results The AT1R [(10.55 ±0.31)% vs (7.04 ±0.11)%] and HIF-1α [(10.51 ±0.52)% vs (8.96 ±0.31)%] in the glomeruli of typeⅠLN was significantly higher than healthy controls(all P<0.05). In the early phase of LN, RAS was activated and tissues were ischemic and hypoxic. The highest expression of AT1R (18.22 ± 2.11)% and HIF-1α (19.48 ±0.61)% in glomeruli was found in type Ⅳ LN, especially in juxtaglomerular apparatus, AT1R (19.98 ±0.21)% and HIF-1α(24.90 ±0.70)%. AT1R was positively correlated with HIF-1αin the glomer-ulus (r=0.949, P<0.01) and juxtaglomerular apparatus (r=0.762, P<0.05). AT1R and HIF-1αin juxtaglomerular apparatus was positively correlated with 24-hour proteinuria (r=0.756, P<0.05 and r=0.802, P<0.05). Conclusion High expressions of AT1R and HIF-1α have been shown in active LN biopsies. It proves that RAS is activated by ischemia and hypoxia, then it up-regulates HIF-1α expression. Our results suggest that the two factors may be associated with disease activity of LN.

Brain renin-angiotensin system: a potential therapeutic target for ischemic stroke / 国际脑血管病杂志

Brain renin-angiotensin system (RAS) is closely associated with many pathophysiological processes of cardiocerebrovascular diseases,including stroke.The activation of the different components in RAS will produce specific biological effects.This article reviews the roles of brain RAS in the pathophysiological processes of ischemic stroke,especially the neuroprotective effect of ACE2/Ang-(1-7)/Mas axis.

Progress of the relationship between angiotensin Ⅱ and its type 1 receptor and some malignant tumors / 肿瘤研究与临床

Angiotensin Ⅱ (Ang Ⅱ) is the main effector of the renin-angiotensin system (RAS). As a major regulator of blood pressure and cardiovascular homeostasis, Ang Ⅱis involved in the regulation of cell growth, proliferation and apoptosis. Ang Ⅱtype 1 receptor (AGTR1) is the important part of the RAS by mediating most of the Ang Ⅱ actions. Recently evidence suggested that AGTR1 correlated with tumor angiogenesis and poor patient outcome in cancer. Therefore, AGTR1 blockers have the potential to suppress the tumor angiogenesis and metastasis. This article intents to summarize the progression of the relationship between AngⅡ,AGTR1 and some malignant tumors.

The expressions and clinical relevance of angiotensin-receptor-1 and hypoxia-inducible factor-1α in glomerulus and juxtaglomerular apparatus of patients with lupus nephritis / 中华风湿病学杂志

Objective To detect the expressions of angiotensin-receptor-1 (AT1R) and hypoxia-inducible factor (HIF)-1αin glomeruli and juxtaglomerular apparatus of different types of lupus nephritis (LN) patients, and analyze the correlation between them with systemic lupus erythematosus disease activity index (SLEDAI) complement 3, serum creatinine and 24-hour proteinuria in order to explore the role of the two factors in the pathogenesis of lupus nephritis (LN). Methods Between May 2010 and April 2016, a total of 90 patients with LN and 8 healthy controls were selected from Department of Rheumatology, Qujing Affiliated Hospital of Kunming Medical University and the First Affiliated Hospital of Kunming Medical University. The expressions of AT1R and HIF-1αin renal biopsy specimens were measured by streptavidin-perosidase (SP) of immunohistochemical stains. Pathological graphic analysis system was used for semi-quantitative estimate. Levels of SLEDAI, C3, serum creatinin and 24-hour proteinuria were also detected. Finally the relationshipbetween the two factors with clinical data was analyzed. The ANOVA test was used for intergroup comparison, and SNK-q test was used for the two groups comparison. Pearson's analysis was used for correlation analysis. Results The AT1R [(10.55 ±0.31)% vs (7.04 ±0.11)%] and HIF-1α [(10.51 ±0.52)% vs (8.96 ±0.31)%] in the glomeruli of typeⅠLN was significantly higher than healthy controls(all P<0.05). In the early phase of LN, RAS was activated and tissues were ischemic and hypoxic. The highest expression of AT1R (18.22 ± 2.11)% and HIF-1α (19.48 ±0.61)% in glomeruli was found in type Ⅳ LN, especially in juxtaglomerular apparatus, AT1R (19.98 ±0.21)% and HIF-1α(24.90 ±0.70)%. AT1R was positively correlated with HIF-1αin the glomer-ulus (r=0.949, P<0.01) and juxtaglomerular apparatus (r=0.762, P<0.05). AT1R and HIF-1αin juxtaglomerular apparatus was positively correlated with 24-hour proteinuria (r=0.756, P<0.05 and r=0.802, P<0.05). Conclusion High expressions of AT1R and HIF-1α have been shown in active LN biopsies. It proves that RAS is activated by ischemia and hypoxia, then it up-regulates HIF-1α expression. Our results suggest that the two factors may be associated with disease activity of LN.

Association of polymorphisms in angiotensin Ⅱ receptor gene with primary aldosteronism / 中国基层医药

Objective To investigate the relationship between the polymorphisms of angiotensin Ⅱ receptor gene and the risk of primary aldosteronism (PA).Methods Polymerase chain reaction -restriction fragment length polymorphism (PCR -RFLP)was used to examine the 1 1 66A /C polymorphism of AT1 R gene and 1 675A /G poly-morphism of AT2R gene in 85 patients with PA and 1 00 healthy controls.Results There was no significant difference of AT1 R 1 1 66A /C genotypes (AA,AC,CC)and allele (A and C)frequency among patients and controls (χ2 =0.430,P =0.806).There was obvious difference of AT2R 1 675A /G genotypes (AA,AG,GG)and allele (A and G) frequency among two groups (χ2 =6.1 21 ,P =0.01 3).The G allele was higher than A allele in PA group (χ2 =6.767,P =0.009).Conclusion Homogenic mutation of 1 675A /G site in AT2R gene may be one of risk factors of PA.

Effect of long-acting nitrate on expression of Ang Ⅱ receptor subtypes in kidneys of heart failure rats / 中华老年心脑血管病杂志

Objective To study the effect of long‐acting nitrate on cardiac function and expression of AngⅡreceptor (ATR)subtypes in kidneys of chronic heart failure (CHF)rats after myocardial infarction .Methods Ninety male Wistar rats aged 10 weeks were randomly divided into control group (group A ,n=9) ,sham operation group (group B ,n=8) ,HF model group (group C ,n=9) , low Elantan dose group (group D ,n=9) ,high Elantan dose group (group E ,n=9) ,olmesartan group (group F ,n=9) ,and combined high Elantan dose and olmesartan group (group G ,n=8) .A HF model was established by ligating the left anterior descending artery .The animals received gastric drugs for 6 weeks .Their cardiac function was assyed by ultrasound echocardiography and expressions of AT1 R and AT2 R were detected by RT‐PCR and Western blot ,respectively .Results The PRA and AngⅡexpression levels were significantly higher ,the AT1 R expression level was significantly higher and the AT2 R expression level was significantly lower in group C than in group B (P<0 .01 ,P<0 .05) .The PRA and AngⅡexpression levels were significantly lower ,the AT1 R expression level was significantly lower and the AT2 R expression level was significantly higher in groups E‐G than in group C (P<0 .05 ,P<0 .01) .The receptor expression levels were much higher in group G than in group F (P<0 .05) .Conclusion Long‐term use of long‐acting nitrate can effectively improve cardiac function and protect renal function .

Correlations between AT1R gene polymorphism with AT1R mRNA and protein expression difference in brain tissue after hypertensive cerebral hemorrhage / 重庆医学

Objective To investigate the correlation of angiotensin Ⅱ type 1 receptor (AT1R) gene polymorphism with AT1R expression levels and brain edema after hypertensive intracerebral hemorrhage .Methods 45 operative patients with hypertensive intracerebral hemorrhage in the Affiliated Yongchuan Hospital of Chongqing Medical Univercity from December 2011 to August 2012 were collected as the experimental group and 45 operative patients with refractory epilepsy weres selected as the control group .The venous blood in the two groups were collected for detecting the AT 1R gene polymorphism ;The brain tissue was taken from lesions in operation ,then AT1R mRNA concentration was determined by RT-PCR and the AT1R protein level was determined by Western blot ;Head CT was performed on postoperative 1 ,3 ,5 d;the degree of cerebral edema was reflected by CT value . Results The levels of two kinds of genotype AT1R mRNA in the experimental group had no statistically significant difference(P>0 .05);the operative area CT value of AC genotype was significantly lower than that of AA genotype with statistical difference (P<0 .05);the ATIRmRNA of various genotypes ,protein level and cerebral edema in the control group had no statistical differences . Conclusion The AT 1R gene polymorphism has no obvious correlation with the concentration expression of AT 1R mRNA in the brain tis-sue;there is correlation between AT 1R protein level and AT 1R protein level and the cerebral edema degree in the brain tissue .

Roles of angiotensin Ⅱ receptors in tumor / 国际肿瘤学杂志

Angiotensin (Ang Ⅱ),a main effector peptide of the renin-angiotensin system (RAS),mediates a hormonal action in the maintenance of blood pressure and electrolyte levels,and thus fluid homeostasis.Recent studies have implicated that it correlates with tumor growth,angiogenesis,metastasis and it has drawn more and more attention.Many studies show that Ang Ⅱ-AT1R/AT2R play crucial roles in tumor growth,metastasis,invasion and tumor angiogenesis,which are formed new targets for treating malignant tumors.

Amplification of recombinant adenovirus vector carrying rat angiotensinⅡtype 2 receptor and transfection in INS-1 cells / 军事医学

Objective To amplify the recombinant adenovirus vector carrying rat angiotensin Ⅱ type 2 receptor (AT2R) gene using human embryonic kidney (HEK) 293A cell lines and to construct a pancreatic islet βcell model overexpressing AT2R by transfecting the adenovirus vector into rat insulinoma (INS-1) cell lines.Methods Recombinant adenovirus vector Ad-G-AT2R-EGFP and control vector Ad-CMV-EGFP were amplified with HEK 293A cells and the titer of the adenovirus was detected .After both adenovirus vectors were transfected into INS-1 cells,AT2R and angiotensin Ⅱtype 1 receptor(AT1R) gene expressions were tested using real-time PCR, Western blotting, immunofluorescence staining and confocal laser-scanning microscopy .Results The titer of amplified Ad-G-AT2R-EGFP and Ad-CMV-EGFP was re-spectively 9 ×109 pfu/ml and 8 ×109 pfu/ml.Transfection of Ad-G-AT2R-EGFP into INS-1 cells induced an increase in AT2R mRNA expression in a dose-dependent manner , and significantly increased AT2R mRNA and protein expression compared with Ad-CMV-EGFP-or mock-transfection.Conclusion The recombinant adenoviral vector carrying AT2R gene is successfully amplified and an INS-1 cell model overexpressing AT2R is constructed by transient transfection , which can contribute to further study of the role of AT2R in pancreatic islet βcells.

Relationship of angiotensin Ⅱ type 1 receptor autoantibody and renal cell apoptosis induced by caspase-12 in diabetic nephropathy rats / 中华肾脏病杂志

Objective To study the relationship of angiotensin Ⅱ type 1 receptor (AT1R) autoantibody (AT1-AA) and renal cell apoptosis induced by caspase-12 in diabetic nephropathy (DN)rats.Methods High-sucrose and high-fat diet and intraperitoneal injection of streptozotocin (35 mg/kg) were utilized to establish DN rat model.Serum AT1-AA was detected by enzyme-linked immunosorbent assay (ELISA) and renal cell apoptosis was detected by TUNEL staining.Furthermore,the mRNA levels of the endoplasmic reticulum stress (ERS) chaperone protein glucose regulated protein 78 (GRP78) and ERS-associated apoptosis protein caspase-12 were measured by real-time quantitative PCR.Additionally,the levels of GRP78 and caspase-12 protein were measured by Western blotting.Results The renal cell apoptosis rate in DN group was increased significantly (P ＜ 0.01),and the renal cells apoptosis rate in AT1-AA positive DN group was higher than that in AT1-AA negative DN group [(20.05±1.71)％ vs (13.24±4.93)％,P ＜ 0.01].The mRNA expressions of GRP78 and caspase-12 in DN group,in comparison to NC group,were increased significantly (P ＜ 0.01),as well as the proteins (P ＜ 0.01).And the expression of these mRNA and proteins had significant increment in AT1-AA positive DN rats when compared with AT1-AA negative DN rats (P ＜ 0.05).Conclusions AT1-AA can induce ERS in the renal tissue of DN rats,and promote renal cell apoptosis likely via the modulation of caspase-12 signaling pathway.

Expression and significance of autoantibodies against angiotensin Ⅱ type 1 receptor in acute coronary syndrome / 中国医师进修杂志

Objective To examine the expression of autoantibodies against angiotensin Ⅱ type 1 receptor (AT1-AAs),monocyte chemoattractant protein-1 (MCP-1) and high-sensitivity C-reactive protein (hs-CRP) in patients of acute coronary syndrome (ACS),and study the role of AT1-AAs in plaque stability and pathogenesis of ACS.Methods Sixty patients with ACS were selected as ACS group,60 patients with stable angina pectoris (SAP) were selected as SAP group,and 60 healthy people were selected as control groups.The epitopes of the second extracellular loop of angiotensin Ⅱ type 1 receptor (165-191) were synthesized and used as antigen to screen the serum autoantibodies by enzyme-linked immunosorbent assay (ELISA).The peripheral blood levels of MCP-1 and hs-CRP were also evaluated.Results The positive rates of AT1-AAs in ACS group,SAP group and control group were 45.0％(27/60),21.7％(13/60) and 5.0％(3/60),respectively.The positive rates of AT1-AAs in ACS group and SAP group were significantly higher than those in control group,the positive rate of AT1-AAs in ACS group was significantly higher than that in SAP group,and there were statistical differences (P ＜ 0.01).The MCP-1 and hs-CRP levels in ACS group and SAP group were significantly higher than those in control group,the MCP-1 and hs-CRP levels in ACS group were significantly higher than those in SAP group,and there were statistical differences (P ＜ 0.01).The MCP-1 and hs-CRP levels in AT1-AAs positive patients in ACS group and SAP group were significantly higher than those in AT1-AAs negative patients,and there were statistical differences (P ＜0.01).Conclusions AT1-AAs may play an important role in the pathogenesis of ACS.Inducing the expression of inflammatory factor through AT1-AAs maybe an important mechanism for plaque instability.

The changes and the clinical significance of plasma Apelin in elderly patients with sepsis / 中华老年医学杂志

Objective To investigate the clinical characteristic and the predicting value of plasma Apelin in elderly patients with sepsis.Methods A retrospective analysis was conducted in 26 sepsis patients aged (72.9±9.7) years in average and 30 healthy controls.Serum Apelin level was measured by ELISA.Body mass index (BMI) and C-reaction protein (CRP) were detected.Patients were divided into survival group (n=18) and death group (n 8).According to acute physiology and chronic health evaluation (APACHE) Ⅱ,patients were divided into subgroup A (n=14,APACHE Ⅱ score≤20) and subgroup B (n=12,APACHE Ⅱ score＞20).Results The Apelin concentration was higher in sepsis patients than in healthy controls [(0.38±0.15)ng/L vs.(0.19±0.12)ng/L,t=2.011,P＜0.05].The Apelin concentration was lower in survival group than in death group[(0.21 ± 0.29)ng/L vs.(0.49 ± 0.32) ng/L,t =2.094,P＜0.05].The Apelin level was increased with APACHE Ⅱ scores increment in sepsis patients (P＜0.05).Multivariable logistic analysis showed that when taking survival/death as the dependent variable and Apelin as the independent variable,the ()R value was 4.162 with 95％ CI:1.115-15.535(P＜0.05).Conclusions Increased serum Apelin level reflects the severity of illness in patients with sepsis,which is a risk factor for death in prognosis of sepsis.

Regulation of human G-coupled protein kinase 4 A142V on AT1 receptor in rat vascular smooth muscle cells / 重庆医学

Objective To study the effect of human G-coupled protein kinase 4(GRK4) A142V overexpression on angiotensin Ⅱ1 type(AT1 ) receptor and its-mediated proliferation of rat vascular smooth muscle cells .Methods We constructed a lentiviral vec-tor carrying human GRK4-EGFP gene and observed its expression in A10 cells .Expression of AT1 receptor were determined by im-munoblotting ,GRK4 activity were checked by spectrophotometry ;the linkage between GRK4 and AT1 receptor were determined by co-immunoprecipitation .[3 H] thymidine incorporation was used to detect changes of cell proliferation .Results As compared with the control cells ,A142V-transfected cells had higher GRK4 activity and higher AT1 receptor expression ;there was linkage between GRK4 and AT1 receptor ,the co-immunoprecipitation levels were lower in A142V cells .The basal levels of VSMC proliferation was higher in A142V cells ,Ang Ⅱ increased VSMC proliferation to a greater extent in A 142V cells .Conclusion GRK4 A142V ,via in-creasing GRK4 activity ,increases AT1 receptor expression and function in vascular smooth muscle cell proliferation .

Correlation between renin-angiotensin-aldosterone system gene polymorphisms and large artery atherosclerotic stroke: a study in a southem Chinese Han population / 国际脑血管病杂志

Objective To study the correlation between the renin-angiotensin-aldosterone system angiotensinogen (AGT) gene M235T,angiotensin Ⅱ type 1 receptor (AGTR1) gene Al166C,aldosterone synthase (CYP11B2) gene -344C/T polymorphisms and large-artery atherosclerotic (LAA) stroke in a southern Chinese Han population.Methods Polymerase chain reaction and gene sequencing technology were used for the genotyping in patients with LAA and normal controls with AGT gene M235T,AGTR1 gene A1166C,and CYP11B2 gene - 344C/T polymorphisms in a southern Chinese Han population,and to determine the correlation between the 3 gene polymorphisms and LAA by binary logistic regression analysis.Results A total of 107 patients with LAA and 142 healthy controls were included in the study.The frequencies of the AGT gene 253TT genotype (66.36％ vs.50.70％,x2 =6.122,P =0.047) and T allele (79.44％ vs.70.07％ ％,x2 =5.581,P =0.018) in the LAA group were significantly higher than those in the control group.The frequencies of the AGTR1 gene 1166CC genotype (0％ vs.0％,x2 =1.494,P =0.222) and C allele (7.48％ vs.4.93％,x2 =1.399,P =0.237) in the LAA group were no significantly differences with those in the control group.The frequencies of the CYP11B2 gene - 344CC genotype (9.35％ vs.4.23％,x2 =3.603,P =0.165) and C allele (27.10％ vs.26.06％,x2 =0.069,P =0.793) in the LAA group were no significant differences with those in the control group.Binary logistic regression analysis showed that there was no significant correlation between the three gene polymorphisms and the simple LAA diseases.The frequencies of AGT gene 235TT genotype (68.00％ vs.41.90％,x2 =12.446,P =0.002) and T allele (79.33％ vs.64.76％,x2 =8.993,P =0.003) in the LAA patients complicated with hypertension were significantly higher than those in the normotensive control group.Logistic regression analysis showed that the odds ratio (OR) exposed to TT genotype was 2.153 (95％ confidence interval [CI] 0.789-5.872).The OR of T allele was 2.089 (95％ CI 1.285-3.396).Conclusions The AGT gene M235T polymorphism is not associated with the simple LAA in the southern Chinese Han population,but it may be associated with the risk of LAA complicated with hypertension;CYP11B2 gene -344C/T polymorphism and AGTR1 gene A1166C polymorphism are not associated with the onset of LAA in the southern Chinese Han population.

Expression and significance of AT1-AA and ET1 in materal peripheral blood, umbilical cord blood and placenta in preeclampsia / 中华妇产科杂志

Objective To investigate the expression of autoantibodies to the angiotensin Ⅱ type Ⅰreceptor (AT1-AA) and endothelin-1 (ET-1) in pregnant women's blood and explore their correlation with the pathogenesis of preeclampsia.Methods Ninety pregnant women who delivered from June 2011 to December 2011 in the First Affiliated Hospital of Zhengzhou University were chosen as the study objects.They were divided into mild preeclampsia group (n =30),severe preeclampsia group (n =30) and normal group (control group,n =30).The levels of AT1-AA and ET1 in maternal peripheral blood and umbilical cord blood were detected by ELISA,and the mRNA expression levels of AT1-AA and ET1 in placenta tissues were determined by reverse transcription (RT) PCR.Moreover,the correlation clinical indexes were detected and analysed.Results (1) The levels of AT1-AA and ET1 in maternal peripheral blood of preeclampsia [mild group:(114 ± 19) ng/L and (31 ± 9) ng/L,severe group:(145 ± 15) ng/L and (38 ± 10) ng/L] were both significantly higher than that of control group [(59 ± 5) ng/L,(17 ±4) ng/L].In addition,compared with mild group,the levels of AT1-AA and ET1 in severe group were significantly higher (P ＜0.05).(2) The levels of AT1-AA and ET1 in umbilical cord blood of preeclampsia [mild group:(105 ± 14) ng/L and (35 ±6) ng/L,severe group:(118 ± 14) ng/L and (40 ±5) ng/L] were significantly higher than that of control group [(61 ± 12) ng/L,(24 ± 5) ng/L].In addition,compared with mild group,the levels of AT1-AA and ET1 in severe group were significantly higher (P ＜0.05).(3) The mRNA expression levels of AT1-AA and ET1 in placenta tissues of mild group (0.313 ± 0.039,0.296 ±0.028) and severe group (0.568 ±0.052,0.577 ±0.046) were significantly higher than that in control group (0.198 ± 0.017,0.137 ± 0.012),and the levels in severe group were significantly higher than that in mild group (P ＜0.05).(4) There was an evident positive correlation between AT1-AA and ET1 levels of preeclampsia women's peripheral blood,umbilical cord blood and placenta (P ＜ 0.05).(5) The level of AT1-AA in umbilical cord blood of preeelampsia pregnant women was positively correlated with S/D value of umbilical artery (P ＜ 0.05),and negatively correlated with the weight of the birth and the placental (P ＜ 0.05).Conclusion The AT1-AA in the blood of pregnant women plays an important role in promoting the generation and development of preeclampsia by increasing the ET1 secretion.

Effects of cytokines on expression of angiotensin Ⅱ type 1 receptors in vascular smooth muscle cells in rats / 中华麻醉学杂志

Objective To investigate the effects of cytokines on the expression of angiotensin Ⅱ type 1 receptor (AT1R) in vascular smooth muscle cells (VSMCs) in rats. Methods Primary cultured VSMCs from SD rat thoracic aorta were cultured in serum-free DMEM for 24 h, and then in DMEM supplemented with 10% fetal bovine serum for another 12 h. The cultured VSMCs were randomly divided into 5 groups (n =6 each): control group (group C); 10% cytokine group (group L); 50% cytokine group (group N); 100% cytokine group (group H) and L-arginine methy ester (L-NAME), an inhibitor of nitric oxide synthase) group. In group C, the cellswere cultured continuously for 12 h. In L, N and H groups, 10%, 50% and 100% cytokines (IL-1β 50 ng/ml +TNF-α 100 ng/ml + IFN-γ 500 ng/ml) were added to the culture medium respectively and the cells were then incubated for 12 h. In group L-NAME, 100% cytokines + L-NAME 5 mmol/L were added to the culture medium and the cells were then incubated for 12 h. The expression of AT1R mPNA and protin was determined by RT-PCR and Western blot respectively.Results Cytokines down-regulated AT1R mRNA and protein expression in a concentration-dependent manner (P ＜ 0.05 or 0.01). L-NAME reversed cytokines-induced changes in AT1R mRNA and protein expression ( P ＜ 0.01). Conclusion Cytokines can down-regulate the expression of AT1R in rat VSMCs and the mechanism is related to the NO synthesis.