Preoperative angiotensin II type 2 receptor is a predictor for developing chronic post-surgical pain after total knee arthroplasty surgery.

OBJECTIVE: This study aimed to explore whether preoperative angiotensin II type 2 receptor (AT2R) level in knee osteoarthritis (OA) patients was an independent risk factor for chronic post-surgical pain (CPSP) after total knee arthroplasty (TKA). METHODS: A total of 220 patients who had undergone unilateral TKA were enrolled from October 2019 to January 2020. Quantitative sensory testing (QST), PainDETECT questionnaires (PD-Q), the Western Ontario McMaster Universities Osteoarthritis Index (WOMAC), the hospital anxiety and depression (HAD) and serum AT2R were collected preoperatively. The primary outcome was the incidence of CPSP, which was defined as the visual analogue scale (VAS) score ≥ 4 in the ipsilateral knee joint six months after operation. RESULTS: The prevalence of CPSP was 13.6% (n = 30). Multiple logistic regression analysis showed that patients with higher AT2R level (OR: 1.007, 95% CI: 1.003-1.011) and PD-Q score (OR: 1.146, 95% CI: 1.008-1.298) before surgery had an increased risk of CPSP after surgery, and a combination of preoperative AT2R and PD-Q (Akaike information criterion: 147.2; area under receiver operating characteristic (ROC) curve: 0.890) was able to correctly classify 90.16% of patients into CPSP positive or negative groups. CONCLUSION: Our findings suggest that patients with higher preoperative AT2R level are at increased risk of developing CPSP following TKA. AT2R may serve as a candidate predictor for phenotyping CPSP in OA patients.

Circulating angiotensin type II receptor: Possible marker for antibody mediated rejection after renal transplantation?

BACKGROUND: Presence of antibody [Ab] against angiotensin receptor [AT1R] indicates heightened risk for antibody mediated rejection [AMR] after transplantation but is insufficient as a marker. We speculated AT1R might be released systemically because of AMR and might be a useful biomarker. METHODS: AT1R was measured in blood from 73 Normals and 72 renal patients pre- and post-transplantation. Patients were stratified as AMR-free [Gp1], AMR<1yr [Gp2] and AMR>1yr [Gp3]. RESULTS: AT1R was higher [13±26vs.367±537, p<0.01)] and more prevalent [20% vs. 92%, p<0.01] among renal patients than Normals. Pretransplant levels were similar [p=ns] between groups. One-year posttransplant levels approached [p<0.01] normalcy for Gps1+3 but spiked during AMR and remained elevated [155±58, p<0.01] for 50% Gp2 patients. One-year AT1R levels were higher among subsequent graft failures than surviving grafts [171±267vs. 38±50, p<0.01]. CONCLUSIONS: Pretransplant AT1R was abnormally elevated: possibly indicating ongoing tissue injury. Pretransplant AT1R didn't predict risk for AMR. However, AT1R spiked during early AMR and sustained elevations were associated with poorer outcomes.

Compound 21, a selective angiotensin II type 2 receptor agonist, downregulates lipopolysaccharide-stimulated tissue factor expression in human peripheral blood mononuclear cells.

Intricate interrelationships connect tissue factor (TF), the principal initiator of the clotting cascade, to inflammation, a cross-talk amplified by locally active angiotensin II, a proinflammatory agent with direct TF-stimulating properties mediated by the angiotensin II type 1 receptor (AT1R)s. However, angiotensin II also stimulates angiotensin II type 2 receptor (AT2R)s and they may as well contribute to TF expression, a possibility in need of further evaluation. We investigated the effect of C21, a highly specific AT2R agonist, on TF antigen (ELISA), procoagulant activity (PCA, one-stage clotting assay) and TF-mRNA (real-time PCR) in peripheral blood mononuclear cell (PBMC)s activated by lipopolysaccharide (LPS), a pro-inflammatory and procoagulant stimulus. C21 downregulated LPS-stimulated TF antigen, PCA and TF mRNA, an effect abolished by PD123 319, a selective AT2R antagonist, and left unchanged by omesartan, a selective AT1R antagonist. PD123 319 per se did not affect LPS-induced TF expression while omesartan inhibited and BAY 11-7082, a specific NFκB inhibitor, abolished endotoxin-activated procoagulant activity (PCA). C21, a selective AT2R agonist, downregulates the transcriptional expression of TF in LPS-activated PBMCs, a finding consistent with the existence in PBMCs of AT2Rs whose stimulation attenuates inflammation-mediated procoagulant responses. The data open insofar unexplored and potentially relevant facets to our understanding of the complex links connecting angiotensin II to inflammation and coagulation.

Angiotensin II induces the generation of procoagulant microparticles by human mononuclear cells via an angiotensin type 2 receptor-mediated pathway.

INTRODUCTION: Microparticles are small vesicles shed by cells upon activation and during apoptosis which participate in physiologically relevant phenomena, including blood coagulation. Intracellular calcium mobilization is one of the mechanisms of microparticle generation during cell activation. Because the renin-angiotensin system has been proposed as a link between hypertension and increased thrombotic risk, we investigated whether angiotensin II upregulates the generation of procoagulant microparticles by human mononuclear cells. MATERIALS AND METHODS: Human mononuclear cells were exposed to angiotensin II for 15min. Intracellular calcium concentration was assessed by a Fluo 4 based kit. The supernatants were analyzed for both microparticle content, with a commercially available kit based on phosphatidylserine analysis, and microparticle-associated tissue factor, with a one-stage clotting assay. RESULTS: Intracellular calcium concentration is increased upon exposure of mononuclear cells to angiotensin II. Incubation with angiotensin II stimulates microparticles release; microparticle-associated tissue factor is also upregulated. The effect is inhibited by an angiotensin receptor type 2 antagonist (PD123319) and not by two angiotensin type 1 antagonists (Losartan and Olmesartan). CONCLUSIONS: Angiotensin receptor 2-mediated upregulation of tissue factor-bearing, procoagulant microparticle generation represents a novel mechanism linking the renin-angiotensin system to thrombosis.

Angiotensin 1-7 and Mas decrease thrombosis in Bdkrb2-/- mice by increasing NO and prostacyclin to reduce platelet spreading and glycoprotein VI activation.

Bradykinin B2 receptor-deleted mice (Bdkrb2(-/-)) have delayed carotid artery thrombosis times and prolonged tail bleeding time resulting from elevated angiotensin II (AngII) and angiotensin receptor 2 (AT2R) producing increased plasma nitric oxide (NO) and prostacyclin. Bdkrb2(-/-) also have elevated plasma angiotensin-(1-7) and messenger RNA and protein for its receptor Mas. Blockade of Mas with its antagonist A-779 in Bdkrb2(-/-) shortens thrombosis times (58 ± 4 minutes to 38 ± 4 minutes) and bleeding times (170 ± 13 seconds to 88 ± 8 seconds) and lowers plasma nitrate (22 ± 4 µM to 15 ± 5 µM), and 6-keto-PGF1α (259 ± 103 pg/mL to 132 ± 58 pg/mL). Bdkrb2(-/-) platelets express increased NO, guanosine 3',5'-cyclic monophosphate, and cyclic adenosine monophosphate with reduced spreading on collagen, collagen peptide GFOGER, or fibrinogen. In vivo A-779 or combined L-NAME and nimesulide treatment corrects it. Bdkrb2(-/-) platelets have reduced collagen-related peptide-induced integrin α2bß3 activation and P-selectin expression that are partially corrected by in vivo A-779, nimesulide, or L-NAME. Bone marrow transplantations show that the platelet phenotype and thrombosis time depends on the host rather than donor bone marrow progenitors. Transplantation of wild-type bone marrow into Bdkrb2(-/-) hosts produces platelets with a spreading defect and delayed thrombosis times. In Bdkrb2(-/-), combined AT2R and Mas overexpression produce elevated plasma prostacyclin and NO leading to acquired platelet function defects and thrombosis delay.

Association between angiotensin II Type 2 receptor gene A/C3123 polymorphism and high-density lipoprotein cholesterol with hypertension in asymptomatic women.

OBJECTIVE: The present study investigated the association between the angiotensin II type 2 receptor (AT2R) gene adenine/cytosine (A/C)-3123 polymorphism and cardiometabolic variables in subjects with and without hypertension. METHODS: Cardiometabolic variables, in addition to genotyping by an allele-specific DNA assay, were measured in 161 asymptomatic community-dwelling Japanese women (age range 30-83 years). They were divided into hypertensive (n = 82, age 50-81 years) and nonhypertensive (n = 79, age 30-83 years) subjects. RESULTS: The A-allele carriers (n = 53) showed significantly lower high-density lipoprotein cholesterol (HDL-C) levels than the non-A-allele carriers (n = 26) among nonhypertensive subjects (1.45 ± 0.38 vs. 1.66 ± 0.33 mmol/l, p = 0.02). Even when multiple-adjusted analyses were performed, the HDL-C levels continued to differ significantly and independently of other variables, including the body mass index and insulin resistance index, between A-allele and non-A-allele carriers. However, this association was not observed among hypertensive subjects. CONCLUSION: The present study demonstrated that A-allele carriers had significantly lower HDL-C levels than did non-A-allele carries among nonhypertensive women, while this association was not observed among hypertensive women. This indicates that the A/C3124 polymorphism may be a marker associated with HDL metabolism by hypertension. This was a small study, so further research is warranted to confirm the observed association.

Angiotensin II type 1 and type 2 receptor expression in circulating monocytes of diabetic and hypercholesterolemic patients over 3-month rosuvastatin treatment.

BACKGROUND: In diabetes, a variety of pro-inflammatory cellular changes has been found in various cell types, including monocytes which are known to be involved in all the phases of atherogenesis. Angiotensin II (Ang II) type 1 receptor (AT1R) mediates the pro-atherogenic effects of Ang II whereas the type 2 receptor (AT2R) seems associated with atheroprotection. We sought to investigate the potential changes of AT1R-AT2R expression in human monocytes of type 2 diabetic- hypercholesterolemic patients and in hypercholesterolemic subjects, upon clinical treatment with rosuvastatin. METHODS: The AT1R membrane protein and mRNA AT1R and AT2R expression in monocytes were investigated in 10 type 2 diabetic-hypercholesterolemic patients and in 10 hypercholesterolemic subjects, before and after 3-month rosuvastatin treatment. Moreover, the serum cytokine levels of interferon-γ (IFN-γ) and interleukin-4 (IL-4) were detected. RESULTS: As expected, rosuvastatin was associated with a change in the lipid profile in the two groups. Both the membrane protein (P = 0.008) and the AT1R mRNA expression (P = 0.038) were significantly reduced during treatment in the absence of AT2R expression change in diabetic-hypercholesterolemic patients whereas no significant difference was observed in hypercholesterolemic subjects. The serum IL-4 levels were increased during treatment whereas no change was observed in IFN-γ in diabetic-hypercholesterolemic patients. No cytokine change was observed in hypercholesterolemic subjects. CONCLUSIONS: Our study on monocytes of diabetic-hypercholesterolemic patients, showing a reduced AT1R but not AT2R expression during rosuvastatin treatment, suggests that statin therapy may modulate favorably the AT1-AT2 receptor balance in subjects with coexistent type 2 diabetes.

Angiogenic factors and their soluble receptors predict organ dysfunction and mortality in post-cardiac arrest syndrome.

INTRODUCTION: Post-cardiac arrest syndrome (PCAS) often leads to multiple organ dysfunction syndrome (MODS) with a poor prognosis. Endothelial and leukocyte activation after whole-body ischemia/reperfusion following resuscitation from cardiac arrest is a critical step in endothelial injury and related organ damage. Angiogenic factors, including vascular endothelial growth factor (VEGF) and angiopoietin (Ang), and their receptors play crucial roles in endothelial growth, survival signals, pathological angiogenesis and microvascular permeability. The aim of this study was to confirm the efficacy of angiogenic factors and their soluble receptors in predicting organ dysfunction and mortality in patients with PCAS. METHODS: A total of 52 resuscitated patients were divided into two subgroups: 23 survivors and 29 non-survivors. The serum levels of VEGF, soluble VEGF receptor (sVEGFR)1, sVEGFR2, Ang1, Ang2 and soluble Tie2 (sTie2) were measured at the time of admission (Day 1) and on Day 3 and Day 5. The ratio of Ang2 to Ang1 (Ang2/Ang1) was also calculated. This study compared the levels of angiogenic factors and their soluble receptors between survivors and non-survivors, and evaluated the predictive value of these factors for organ dysfunction and 28-day mortality. RESULTS: The non-survivors demonstrated more severe degrees of organ dysfunction and a higher prevalence of MODS. Non-survivors showed significant increases in the Ang2 levels and the Ang2/Ang1 ratios compared to survivors. A stepwise logistic regression analysis demonstrated that the Ang2 levels or the Ang2/Ang1 ratios on Day 1 independently predicted the 28-day mortality. The receiver operating characteristic curves of the Ang2 levels, and the Ang2/Ang1 ratios on Day 1 were good predictors of 28-day mortality. The Ang2 levels also independently predicted increases in the Sequential Organ Failure Assessment (SOFA) scores. CONCLUSIONS: We observed a marked imbalance between Ang1 and Ang2 in favor of Ang2 in PCAS patients, and the effect was more prominent in non-survivors. Angiogenic factors and their soluble receptors, particularly Ang2 and Ang2/Ang1, are considered to be valuable predictive biomarkers in the development of organ dysfunction and poor outcomes in PCAS patients.

Gene polymorphisms of ACE and the angiotensin receptor AT2R1 influence serum ACE levels in sarcoidosis.

Angiotensin converting enzyme (ACE) is thought to influence susceptibility, disease progression, and/or outcome of sarcoidosis by functional mutations/polymorphisms of the ACE gene, such as the ACE gene deletion/insertion (D/I) polymorphism or the angiotensin receptors like the angiotensin II receptor type 1 (AT2R1) A1166 --> C polymorphism. The aim of our study was to examine the distribution of the ACE D/I genotypes and the AT2R1 A1166 --> C genotypes in sarcoidosis and healthy controls, and to test their influence on disease progression. In this study, we assessed ACE and AT2R1 genotypes by PCR in 264 healthy Caucasians and 95 sarcoidosis patients. Serum ACE levels were determined using a kinetic test. Genotyping sarcoidosis patients for the AT2R1 A166 --> C polymorphism revealed an increase in homozygous genotypes CC (sarcoidosis: 11.6%, controls: 9.2%) and AA (sarcoidosis: 61.1%, controls: 47.3%) but a lower frequency in heterozygous genotypes (sarcoidosis: 27,4%, controls: 43,5%; p = 0.024) which was more pronounced in male patients. The co-incidence of DI and AC was less frequent in patients with sarcoidosis, suggesting protection by the combination of DI and AC. The AT2R1 A1166 --> C gene polymorphism modulated the effect of the ACE D/I polymorphism on serum ACE levels with the A allele promoting its influence and the C allele reducing it. We conclude that neither the ACE D/I nor the AT2R1 A1166 --> C polymorphism has a role in sarcoidosis disease progression. In males, the homozygous AT2R1 genotypes CC and AA possibly increase the risk for sarcoidosis. Co-incidence of the heterozygous genotypes DI and AC might be protective against sarcoidosis.

C-reactive protein causes downregulation of vascular angiotensin subtype 2 receptors and systolic hypertension in mice.

BACKGROUND: Chronic elevations in circulating C-reactive protein (CRP) are associated with a greater risk of hypertension. Whether elevations in CRP cause hypertension is unknown. METHODS AND RESULTS: Chronic, conscious blood pressure (BP) measurements were performed by radiotelemetry in wild-type CF1 control and CF1 transgenic mice expressing rabbit CRP (CF1-CRP) under the regulation of the phosphoenolpyruvate carboxykinase promoter. Compared with controls, CF1-CRP mice had hypertension that was predominantly systolic, and the severity of hypertension varied in parallel with changes in CRP levels modulated by dietary manipulation. Mice that were hemizygous for the transgene with CRP levels of 9 microg/mL were also hypertensive, indicating that modest elevations in CRP are sufficient to alter BP. CRP transgenic mice had exaggerated BP elevation in response to angiotensin II and a reduction in vascular angiotensin receptor subtype 2 (AT2) expression. In contrast, the decline in BP with angiotensin receptor subtype 1 (AT1) antagonism and vascular AT1 abundance were unaltered, which indicates a selective effect of CRP on AT2. Ex vivo experiments further showed that the CRP-induced decrease in AT2 is a direct effect on the vascular wall, not requiring systemic responses, and that it is reversed by an NO donor, which indicates a role for NO deficiency in the process. In parallel, the chronic inhibition of NO synthase in wild-type mice attenuated vascular AT2 expression without affecting AT1. CONCLUSIONS: These findings provide direct evidence for CRP-induced hypertension, and they further identify a novel underlying mechanism involving downregulation of AT2 related to NO deficiency.

Long-term prevention of hypertension and end-organ damage in Ren-2 transgenic rats is achieved only with persistent but not transient AT1 receptor blockade.

The aim of this study was first to evaluate the effects of persistent or transient blockade of the angiotensin II (ANG II) receptor AT(1) on the development of hypertension and end-organ damage in hypertensive Ren-2 transgenic rats (TGR), and second to assess the potential role of AT(2) receptors in the control of blood pressure (BP) in this monogenetic model of hypertension. Male heterozygous TGR and Hannover Sprague-Dawley (HanSD) rats fed a normal salt diet were treated from day 32 of age either persistently until the end of the experiment (day 100 of age) or transiently until day 56 of age with the selective AT(1) receptor antagonist candesartan or with the combination of candesartan and the AT(2) receptor antagonist PD 123319. Persistent treatment with candesartan completely prevented the rise in BP, proteinuria and the increase in left ventricular weight/body weight ratio, whereas transient treatment with candesartan was effective only as long as the drug was administered. In the presence of candesartan, PD 123319 was without effect. Our results show that in male heterozygous TGR persistent candesartan treatment completely prevented hypertension and end-organ damage as long as the drug was administered, whereas transient AT(1 )receptor blockade had no long-term effects.

[Differences of Angiotensin II receptor expression between systemic and pulmonary circulation in children with left-to-right shunt congenital cardiac lesions].

OBJECTIVE: It has been shown that angiotensin converting enzyme inhibitors (ACEI) can reduce the ratio of pulmonary and systemic circulation blood flow (Qp/Qs) and thus decrease the blood flow of left-to-right shunt in children with left-to-right shunt congenital cardiac lesions. This suggests that there are differences in the expression of Angiotensin II receptors between systemic and pulmonary circulation. This study aimed to explore the differences of Angiotensin II receptors type 1 and type 2 (AT1 and AT2 receptors) expression between systemic and pulmonary circulation in children with left-to-right shunt congenital cardiac lesions. METHODS: Lung and skeletal muscular tissues were obtained from 20 children with left-to-right shunt congenital cardiac lesions by biopsy during operation. The specimens were stained by immunohistochemistry techniques for AT1 and AT2 receptors. The technique of morphometric analysis was used to measure the immunoreactivity of AT1 and AT2 receptors (expressed by IOD values) of pulmonary, skeletal muscular and pleural small vascular wall the diameter of which was 15-100 microm. RESULTS: The immunoreactivities of AT1 and AT2 receptors of pulmonary small vascular walls [(124 +/- 95)x10(3) and (85 +/- 62)x10(3) respectively] were significantly lower than those of skeletal muscular [(219 +/- 156)x10(3) and (155 +/- 139)x10(3) respectively] and those of pleural small vascular walls [(279 +/- 191)x10(3) and (175 +/- 128)x10(3) respectively] in children with left-to-right shunt congenital cardiac lesions (P < 0.05). CONCLUSIONS: The expression of AT1 and AT2 receptors in small vascular walls of systemic circulation was higher than that of pulmonary circulation in children with left-to-right shunt congenital cardiac lesions.