AT1R Expression Research Articles

GW24-e2404 Effect of Prenatal Hypoxia on The Expression of Local Renin - angiotensin System in Cerebral Cortex from Rat Offspring

Objectivesto study the effect of chronic hypoxia during pregnancy on the expression of ACE and the receptors of AngII (AT1R, AT2R) in cerebral cortex from rat offspring.MethodsMaternal hypoxic model was...

GW24-e3170 Angiotensin-(1-7) downregulates angiotensin II type 1 receptor and inhibites mechanical stress-induced cardiomyocyte autophagy

ObjectivesThe renin-angiotensin system peptides are critically involved in the regulation of autophagy. Angiotensin-(1-7) [Ang-(1-7)] has many beneficial effects in the cardiac remodelling. Here, we tested the hypothesis that Ang-(1-7) has...

Angiotensin type 2 receptors in the intermediolateral cell column of the spinal cord: Negative regulation of sympathetic nerve activity and blood pressure

Our previous study demonstrated that AT2R in brainstem nuclei participated in the regulation of sympathetic outflow and cardiovascular function. However, the functional significance of AT2R in the intermediolateral cell column (IML) of the thoracic spinal cord in normal rats remains elusive. We hypothesized that AT2R activation in the IML exerts a sympatho-inhibitory effect. Using Western-blot analysis, immunohistochemical staining and quantitative real-time PCR, both AT1R and AT2R expressions were detected in the spinal cord. The highest AT2R protein expression was found in the IML, while AT1R expression didn't display regional differences within the gray matter. Microinjection of Ang II into the IML dose-dependently elevated mean blood pressure (MAP, employing a transducer-tipped catheter) and renal sympathetic nerve activity (RSNA, using a pair of platinum-iridium recording electrodes), which were completely abolished by Losartan, and attenuated by TEMPOL and apocynin. Activation of AT2R in the IML with CGP42112 evoked hypotension (ΔMAP: -21 ± 4 mmHg) and sympatho-inhibition (RSNA: 73 ± 3% of baseline), which were completely abolished by PD123319 and l-NAME. Blockade of AT2R in the IML with PD123319 significantly increased MAP (11 ± 1 mmHg) and sympathetic nerve activity (RSNA: 133 ± 13% of baseline). Moreover, PD123319 significantly enhanced the Ang II induced pressor response. Furthermore, in isolated IML neurons, CGP42112 treatment augmented potassium current and decreased resting membrane potential by employing whole-cell patch clamp. In the normal condition, AT2R in the IML tonically inhibits sympathetic activity through an NO/NOS dependent pathway and subsequent potassium channel activation.

Expression and significance of ACE2-Ang-(1-7)-Mas axis in the endometrium of patients with polycystic ovary syndrome

To explore the expression of renin-angiotensin system component (ACE-AngII-AT1/AT2 & ACE2-Ang-(1-7)-Mas) in endometrium of polycystic ovary syndrome (PCOS) patients and normal controls. Thirty cases of normal endometrium in proliferative and secretory phases respectively were selected for the protein levels of AngII, AT1, AT2, ACE2, Ang-(1-7) and Mas through immunohistochemistry. And the specimens of proliferative endometrium from 15 PCOS patients and 15 normal controls respectively were prepared for the expressions of AT1, AT2, ACE2 and Mas mRNA through relative quantitative -polymerase chain reaction (RQ-PCR). The histological phases of endometria were confirmed by hematoxylin & eosin staining under microscope. The expressions of AngII, AT1R, AT2R, ACE2, Ang-(1-7) and Mas showed periodical changes in endometrium throughout normal menstrual cycles and shared a similar trend. The expression was more pronounced in epithelial cells than that in stromal cells while it was also more dramatic in secretory phase than proliferative phase; The mRNA expressions of AT1, AT2, ACE2 and MAS were higher in PCOS endometrium than those in normal controls. Statistically significant differences existed between two groups (P < 0.01). There is local existence of RAS in endometrium; Increased expressions of AT1 mRNA, AT2 mRNA, ACE2 mRNA and Mas mRNA in endometrium of PCOS may influence endometrial development and play a role in the pathological process of PCOS.

AB0150 Sex-specific differences in cardiac involvement in ssc patients – relation to angiotensin and endothelin receptors and antibodies?

BackgroundAutopsy studies showed cardiac fibrosis in the majority of patients with systemic sclerosis (SSc). Male SSc patients are at higher risk for left ventricular failure than their female counterparts suggesting...

Myocardial angiotensin II receptor expression in rats of atrial fibrillation and Artemisia annua extract intervention on the expression

Objective Through establishing a rat model of atrial fibrillation, to study myocardial angiotensin type I （AT1R） and type II receptor （AT2R） mRNA expression levels in of the state atrial fibrillation and Artemisia annua extract on its expression. Methods Rat model of atrial fibrillation was established, Artemisia annua extract was used for intervention and captopril was adopted as controls. AT1R, AT2R mRNA and protein expression were observed by PCR and Western-blot technology. Results Compared with the control group （0.36±0.05） , myocardial ATIR mRNA expression was significantly increased in the modelgroup （0.84±0.04） （P〈0.05） . BothArtemisia annua （0.56±0.03） andcaptopril （0.53±0.04） could significantly reduce the myocardial ATIR mRNA expression in the atrial fibrillation rats （P〈0.05） . Captopril showed obvious ATIR mRNA reduction trend, but there was statistical significance compared with Artemisinic extract （P〉0.05）. Artemisinic extract showed no impact on AT2R mRNA expression. Conclusion ATlR was closely related to the incidence of atrial fibrillation. AT1R expression was significantly increased in atrial fibrillation rat. The artemisinic extract can be effectively reduced fibrillation myocardial ATIR expression, which may link with its artemisinic antiarrhythmic mechanism. Key words: Atrial fibrillation; Angiotensin II receptor; AT1R; Artemisiaannua

Decreased MicroRNA Is Involved in the Vascular Remodeling Abnormalities in Chronic Kidney Disease (CKD)

Patients with CKD have abnormal vascular remodeling that is a risk factor for cardiovascular disease. MicroRNAs (miRNAs) control mRNA expression intracellularly and are secreted into the circulation; three miRNAs (miR-125b, miR-145 and miR-155) are known to alter vascular smooth muscle cell (VSMC) proliferation and differentiation. We measured these vascular miRNAs in blood from 90 patients with CKD and found decreased circulating levels with progressive loss of eGFR by multivariate analyses. Expression of these vascular miRNAs miR-125b, miR-145, and miR-155 was decreased in the thoracic aorta in CKD rats compared to normal rats, with concordant changes in target genes of RUNX2, angiotensin II type I receptor (AT1R), and myocardin. Furthermore, the expression of miR-155 was negatively correlated with the quantity of calcification in the aorta, a process known to be preceded by vascular de-differentiation in these animals. We then examined the mechanisms of miRNA regulation in primary VSMC and found decreased expression of miR-125b, 145, and 155 in VSMC from rats with CKD compared to normal littermates but no alteration in DROSHA or DICER, indicating that the low levels of expression is not due to altered intracellular processing. Finally, overexpression of miR-155 in VSMC from CKD rats inhibited AT1R expression and decreased cellular proliferation supporting a direct effect of miR-155 on VSMC. In conclusion, we have found ex vivo and in vitro evidence for decreased expression of these vascular miRNA in CKD, suggesting that alterations in miRNAs may lead to the synthetic state of VSMC found in CKD. The decreased levels in the circulation may reflect decreased vascular release but more studies are needed to confirm this relationship.

Telmisartan inhibits AGE-induced podocyte damage and detachment

Advanced glycation end products (AGE) formed at an accelerated rate under diabetes, could cause podocyte apoptosis, thereby being involved in the development and progression of diabetic nephropathy. Renin–angiotensin system (RAS) plays a role in diabetic nephropathy as well. However, it remains unknown whether there exists a pathophysiological crosstalk between the RAS and AGE in podocyte damage in diabetic nephropathy. Therefore, this study investigated the effects of telmisartan, an angiotensin II (Ang II) type 1 receptor (AT1R) blocker on AGE or Ang II-induced podocyte damage in vitro. We further examined here the effects of AGE on AT1R expression levels in podocytes. AGE or Ang II not only increased DNA damage of podocytes which was evaluated by comet assay, but also induced cell detachment, both of which were significantly blocked by the treatment with telmisartan. AGE significantly increased AT1R levels in podocytes, whereas podocyte Ang II production was modestly stimulated by AGE. Telmisartan alone did not affect the release of lactate dehydrogenase from podocytes. Our present study suggests that AGE could induce podocyte DNA damage and detachment partly via stimulation of the Ang II–AT1R axis, thus providing a novel beneficial aspect of telmisartan in diabetic nephropathy.

Arsenic upregulates the expression of angiotensin II Type I receptor in mouse aortic endothelial cells

Although chronic arsenic exposure is a well-known risk for cardiovascular disease and has a strong correlation with hypertension, the molecular pathogenesis underlying arsenic exposure-induced hypertension remains poorly understood. To delineate the pathogenesis, we examined changes in the mRNA levels of 2 angiotensin II Type I receptor (AT1R) subtypes, AT1AR and AT1BR, in a mouse aortic endothelial cell line, END-D. Quantitative real-time PCR analysis revealed significant increases in the mRNA levels of 2 AT1R subtypes, AT1AR and AT1BR following sodium arsenite (SA) treatment. Flow cytometry analysis revealed that SA increases the generation of reactive oxygen species (ROS) in a dose-dependent manner. In addition, western blot analysis revealed that SA enhances the phosphorylations of c-Jun N-terminal kinases (JNK) and activated protein 1 (AP-1). These phosphorylations were inhibited by N-acetylcysteine (NAC), an anti-oxidant. Finally, SA-induced AT1R expression was found to be prevented both by NAC and specific JNK inhibitor, SP6001325, strongly indicating that AT1R upregulation is a result of the ROS-mediated activation of the JNK signaling pathway. Taken together, our results indicate that arsenic indeed upregulates the AT1R expression, thus highlighting a role of arsenic-induced aberrant AT1R signaling in the pathogenesis of hypertension.

AT1, AT2, and AT1-7 receptor expression in the uteroplacental unit of normotensive and hypertensive rats during early and late pregnancy

IntroductionWe investigated the expression of angiotensin receptors in early pregnancy and established whether normal pregnancy or preeclampsia alters the expression and distribution of the uteroplacental AT1R, AT2R and mas/AT1-7R at late gestation. MethodsThe percentage of each receptor subtype present in tissues from virgin rats and from normotensive and RUPP hypertensive pregnant rats was established by in vitro receptor autoradiography. Receptor mRNA levels were determined by quantitative PCR at early and late pregnancy. ResultsAT1R mRNA levels were up-regulated in the interimplantation (IIS) site at day 7 of gestation. AT2R mRNA levels were decreased at day 5 and 7 in the IIS but increased in the implantation site (IS) at day 5 and 7 as compared to the IIS at day 5. Mas/AT1-7R mRNA was increased in early pregnancy. In normal pregnancy and RUPP the mRNA for all angiotensin receptors was reduced in the uterus at late gestation. The AT1R accounted for the majority of binding in the uterus of virgin and the placenta of pregnant and RUPP. In RUPP pregnancy there was a significant competition with d-Ala in the placenta labyrinth. Discussion and conclusionThe expression of angiotensin receptors suggests their involvement in the maintenance of early stages of pregnancy. During late gestation down-regulation of Ang receptors in the uterus may arise from feedback down-regulation by Ang II. In the placenta the levels of AT1Rs are equivalent in the RUPP model. The increased binding of mas/AT1-7R at late gestation in RUPP may represent a compensatory mechanism to reduce uteroplacental vascular resistance.

Rapamycin Attenuates Cardiac Expression of Anti‐inflammatory AT2R and Anti‐Apoptotic MCL1

Angiotensin II (Ang II) receptor AT2R is a member of the vasodilative/anti‐inflammatory branch of renin‐angiotensin system (RAS). However, mechanisms underlying regulation of AT2R expression in cardiomyocytes are not understood. To determine how vasodilative insulin (INS) and anti‐inflammatory drug rapamycin (Rap) modulate AT2R expression, we performed radioligand binding experiments with 125I‐CGP42112A, an AT2 specific ligand, on mouse atrial cardiomyocyte HL‐1 cells. INS (100nM, 12h) increased expression of functional AT2R that binds 125I‐CGP42112A by 2.2 fold (P&lt;0.05). Co‐treatment with Rap (10nM) attenuated this effect. Our in silico and in vitro studies showed that the AT2R is a target for the diabetic marker microRNA miR‐29 family. Expression levels of miR‐29a, b and c in HL‐1 cells were suppressed by INS and elevated by Rap (~3 fold; P&lt;0.05). Moreover, transfection of HL‐1 cells with miR‐29 mimics reduced 125I‐CGP42112A binding and suppressed insulin‐induced increases in AT2R protein (P&lt;0.05). Immunoblotting showed that Rap also suppressed miR‐29 target Mcl‐1, an anti‐apoptotic gene that promotes cardiomyocyte survival following ischemia‐reperfusion. Collectively these data show for the first time that Rap, a widely used anti‐inflammatory drug, attenuates cardiac expression of AT2R and Mcl1 and up‐regulates miR‐29 family that may promote cardiac INS resistance.

Regulation of Angiotensin II Receptor Expression

The biological functions of angiotensin (Ang) II are mediated by two Ang II receptors, designated type 1 receptor (AT1R) and type 2 receptor (AT2R). Most of the cardiovascular effects of Ang II are mediated by AT1R that is expressed widely in the body. The expression of AT1R is up-regulated in cardiovascular lesions, and is regulated by many endogenous bioactive substances and drugs. The AT2R is generally considered to antagonize the effects of AT1R, but its precise function remains enigmatic and controversial, particularly in humans. The expression of AT2R is low in normal adult animals, but AT2R expression is up-regulated in cardiovascular lesions. The dynamic regulation of AT1R and AT2R expression suggests an active involvement of Ang II receptors in the development of cardiovascular diseases such as atherosclerosis, heart failure, chronic kidney diseases and cerebrovascular diseases. Further clarification of gene regulatory mechanisms of Ang II receptors may identify potential targets for the development of novel therapeutics for cardiovascular diseases.

Central Angiotensin II ‐ induced hypertension downregulates AT2R and the mir‐29 family in rats

Ang II plays a critical role in the pathogenesis of hypertension by enhancing sympathetic outflow via the AT1R. However, the role of AT2R in this process is not clear. The data from our lab revealed an inhibitory influence of AT2R on sympathetic tone. Mir‐29 is predicted to regulate AT2R expression. We hypothesized that the decreased AT2R expression may contribute to hypertension induced by central infusion of Ang II by a mechanism dependent on mir‐29. Male rats received icv infusion of Ang II (50 ng/min) for 7 days. Blood pressure (BP) and heart rate (HR) were measured by telemetry. On day 7, the brain was removed to evaluate protein and mir‐29. As expected, the rats receiving Ang II exhibited an increased BP (38.1 ± 5.2 mmHg) and HR (86.3 ± 11.5 bpm) and an up‐regulated AT1R. In addition, (1) icv Ang II significantly downregulated AT2R protein in PVN and RVLM by 39.2 ± 3.7% and 64.1 ± 4.4%; (2) Gαi2 and nNOS, two downstream signal molecules activated by the AT2R, were significantly down regulated; (3) The expression of mir‐29a and mir‐29b, two potential regulators of AT2R protein, were decreased by 37.5 ± 4.1% and 32.8 ± 2.8%. (4) All of the above alterations were abolished by Losartan. These results suggest that, in the brain, the down regulated AT2R and its downstream signaling pathways contribute to Ang II induced hypertension. The decreased AT2R may be attributable to the inhibition of mir‐29 family by Ang II. Supported by HL093028.

Early changes on proximal tubule Na+‐ATPase activity precede blood pressure elevation and renal dysfunction induced by intrauterine undernutrition: reprogramming by α‐tocopherol

We investigated whether intrauterine undernutrition that induces hypertension and renal dysfunction were preceded by early changes in proximal tubule ouabain‐resistant Na+‐ATPase activity, as well as, whether maternal α‐tocopherol treatment prevents these changes. Male Wistar rats obtained from undernourished dams during gestation presented increased Na+‐ATPase activity in proximal tubule membranes at age of 30 and 60 days. At 30 days of age, PKA activity was lowered, while at 60 days‐old PKC activity was elevated and AT2R expression was reduced. Systolic arterial pressure was increased from age of 105 days and lowered creatinine clearance, increased plasma volume, increased tubulointerstitial ANGII‐positive cells and increased renal p47‐ phox expression was seen at age of 150 days. Maternal α‐tocopherol treatment during lactation recovered these changes. These findings indicate a key role of Na+‐ATPase on blood pressure and renal function modulation, besides to support a non‐antioxidant effect of α‐tocopherol. CAPES, CNPq, FAPERJ, FACEPE.

PP018. Does altitude affect the placental renin angiotensin system (RAS) in pre-eclampsia (PE)?

We have previously shown the activation of placental RAS in high altitude normotensive (NT) pregnancies [1], presumably related to hypoxia during placental development. PE incidence is increased at high altitude; hypoxia-reoxygenation injury contributes to the elevated oxidative stress seen in this condition. Binding of Angiotensin II to its type I receptor (AT1R) increases generation of reactive oxygen species and we have shown an increase in AT1R in PE [2]. Therefore we investigated whether the RAS is further augmented in PE at high altitudes. Biopsies were collected, with informed, written consent, from NT and PE women (ISSHP guidelines) at sea-level (UK) and high altitude, 3100m (Leadville, Colorado) immediately after vaginal delivery or elective non-laboured Caesarean section. mRNA expression of the RAS components (angiotensinogen (AGT), prorenin, prorenin receptor, AT1R and angiotensin type 2 receptor) was determined by qRT-PCR and normalised to 3 stably-expressed housekeeping genes using geNorm. Protein expression was semi-quantitatively assessed by immunohistochemistry (H score) and compared between groups (Mann Whitney U test). mRNA expression of all RAS components was unaffected by altitude in NT, but significantly lower for all in PE (P=0.026-<0.0001). However, in NT, protein expression of all components except AGT was significantly increased at altitude (P=0.037-<0.001). Protein expression of AGT and AT1R was also increased in altitude PE (P<0.004 for both). We have shown the placental RAS to be activated at altitude, with apparent differences in transcription/translation. Hypoxia-inducible transcription factors such as HIF-1 enhance transcription and translation of many genes encoding "rescue" proteins; this might explain the effects of altitude in NTs. HIF-1 is increased in PE, which might allow increased protein expression at altitude in spite of lower mRNA.

Effects of sapindus saponins on inflammatory response mediated by AngⅡ/p38MAPK pathway and cardiac hypertrophy in spontaneously hypertensive rats

To investigate the effects of sapindus saponins on myocardial inflammation mediated by Ang II/ p38MAPK signal pathway and cardiac hypertrophy in spontaneously hypertensive rats. And also to explore the correlation of cardiac hypertrophy and inflammation.Thirty-two 16-week-old spontaneously hypertensive rats (SHR) were randomly divided into four groups, one with placebo as model group, one with captopril tablets (27 mg x kg(-1)) as positive control, one with low-dose sapindus saponins (27 mg x kg(-1)), one with high-dose (108 mg x kg(-1)). And another eight healthy Wistar-Kyoto strain (WKY) rats were used as the normal group. The animals were treated for eight weeks, and the indicators detected were as follows: (1) left ventricular mass index (LVMI); (2) the content of Ang II and hs-CRP in plasma were determined by ELISA; (3) the protein expression of AT1R and VEGF were determined by immunohistochemical method; (4) the protein expression of p-p38MAPK in myocardial cells was determined by Western blot.Sapindus saponins reduced LVMI, and blocked the expression level of Ang II, AT1R, p-p38MAPK, VEGF and hs-CRP in myocardial tissue. Vs the SHR model group, there were significant differences (P < 0.05 or P < 0.01).Our findings suggested that sapindus saponins could inhibited cardiac hypertrophy, the possible mechanisms may be related to the inhibition on inflammatory response mediated by Ang II/p38MAPK pathway.

Exercise Training (ExT) Normalizes Subfornical Organ (SFO)‐ Mediated Sympathoexcitation in Chronic Heart Failure (HF)

ExT normalizes sympathoexcitation (SymEx) exhibited in HF, however the underlying mechanisms are not clear. The SFO is involved in dictating SymEx and is influenced by peptides such as angiotensin (Ang) II due to its weak blood‐brain barrier. We hypothesized that neuronal activity in the SFO would be enhanced in HF and contribute to increased Ang II‐mediated SymEx, and that ExT would reverse this process. HF induced by coronary artery ligation in Sprague‐Dawley rats was confirmed by echocardiography 4 weeks after surgery. HF rats displayed increased neuronal activity in the SFO, as assessed by FosB immunohistochemistry (101 ± 9 vs. 29 ± 2 cells). This increase was attenuated after 4 weeks of treadmill ExT (36 ± 7 cells). In anesthetized rats, microinjection of Ang II into the SFO increased renal sympathetic nerve activity (RSNA) to a greater extent in HF (23 ± 3 vs. 9 ± 1%; 100 pmol), and was normalized after ExT (8 ± 2%). ExT also abolished the decrease in RSNA in HF rats after losartan microinjection into the SFO (−21 ± 4 vs. −1 ± 3%; 200 pmol), implying higher endogenous Ang II tone in HF. ExT reversed the elevated mRNA and protein expression of AT1R in the SFO of rats with HF. The enhanced activation of the SFO by elevated tonic Ang II contributes to the enhanced SymEx exhibited in HF. The reduction in AT1 receptors in the SFO by ExT may be responsible for restoring the neuronal activation in the SFO and SFO‐mediated SymEx. Supp. by NIH HL62222.

Fetal kidney programming by severe food restriction: Effects on structure, hormonal receptor expression and urinary sodium excretion in rats

The present study investigates, in 23-day-old and adult male rats, the effect of severe food restriction in utero on blood pressure (BP), and its association with nephron structure and function changes, angiotensin II (AT1R/AT2R), glucocorticoid (GR) and mineralocorticoid (MR) receptor expression. The daily food supply to pregnant rats was measured and one group (n=15) received normal quantity of food (NF) while the other received 50% of that (FR50%) (n=15). Kidneys were processed to AT1R, AT2R, MR, and GR immunolocalization and for western blotting analysis. The renal function was estimated by creatinine and lithium clearances in 12-week-old offspring. By stereological analyses, FR50% offspring present a reduction of nephron numbers (35%) with unchanged renal volume. Expression of AT1R and AT2R was significantly decreased in FR50% while the expression of GR and MR increased in FR50%. We also verified a pronounced decrease in urinary sodium excretion accompanied by increased BP in 12-week-old FR50% offspring. The current data suggest that changes in renal function are conducive to excess sodium tubule reabsorption, and this might potentiate the programming of adult hypertension. It is plausible to arise in the current study an association between decreasing natriuresis, reciprocal changes in renal AngII and steroid receptors with the hypertension development found in FR50% compared with age-matched NF offspring.

Functionalized dendrimer-based delivery of angiotensin type 1 receptor siRNA for preserving cardiac function following infarction

Cardiovascular disease (CVD) is the leading cause of death throughout the world and much pathology is associated with upregulation of inflammatory genes. Gene silencing using RNA interference is a powerful tool in regulating gene expression, but its application in CVDs has been prevented by the lack of efficient delivery systems. We report here the development of tadpole dendrimeric materials for siRNA delivery in a rat ischemia-reperfusion (IR) model. Angiotensin II (Ang II) type 1 receptor (AT1R), the major receptor that mediates most adverse effects of Ang II, was chosen to be the silencing targeting. Among the three tadpole dendrimers synthesized, the oligo-arginine conjugated dendrimer loaded with siRNA demonstrated effective down-regulation in AT1R expression in cardiomyocytes in vitro. When the dendrimeric material was applied in vivo, the siRNA delivery prevented the increase in AT1R levels and significantly improved cardiac function recovery compared to saline injection or empty dendrimer treated groups after IR injury. These experiments demonstrate a potential treatment for dysfunction caused by IR injury and may represent an alternative to AT1R blockade.

Early Life Stress and Post-Weaning High Fat Diet Alter Tyrosine Hydroxylase Regulation and AT1 Receptor Expression in the Adrenal Gland in a Sex Dependent Manner

Previous studies have shown that early life stress induced by maternal separation or non-handling can lead to behavioural deficits in rats and that these deficits can be alleviated by providing palatable cafeteria high-fat diet (HFD). In these studies we investigated the effects of maternal separation or non-handling and HFD on tyrosine hydroxylase (TH) protein and TH phosphorylation at Ser40 (pSer40TH) and the expression of angiotensin II receptor type 1 (AT1R) protein in the adrenal gland as markers of sympatho-adrenomedullary activation. After littering, Sprague-Dawley rats were assigned to short maternal separation, S15 (15 min), prolonged maternal separation, S180 (180 min) daily from postnatal days 2-14 or were non-handled (NH) until weaning. Siblings were exposed to HFD or chow from day 21 until 19 weeks when adrenals were harvested. Maternal separation and non-handling had no effects on adrenal TH protein in both sexes. We found an effect of HFD only in the females; HFD significantly increased TH levels in NH rats and pSer40TH in S180 rats (relative to corresponding chow-fed groups), but had no effect on AT1R expression in any group. In contrast, in male rats HFD had no effect on TH protein levels, but significantly increased pSer40TH across all treatment groups. There was no effect of HFD on AT1R expression in male rats; however, maternal separation (for 15 or 180 min) caused significant increases in AT1R expression (relative to NH group regardless of diet). This is the first study to report that early life stress and diet modulate TH protein, pSer40TH and AT1R protein levels in the adrenal gland in a sex dependent manner. These results are interpreted in respect to the potential adverse effects that these changes in the adrenal gland may have in males and females in adult life.