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The transition from lean to obese states involves systemic metabolic remodeling that impacts insulin sensitivity, lipid partitioning, inflammation, and glycemic control. Here, we have taken a pharmacological approach to test the role of a nutrient-regulated chromatin modifier, lysine-specific demethylase (LSD1), in obesity-associated metabolic reprogramming. We show that systemic administration of an LSD1 inhibitor (GSK-LSD1) reduces food intake and body weight, ameliorates nonalcoholic fatty liver disease (NAFLD), and improves insulin sensitivity and glycemic control in mouse models of obesity. GSK-LSD1 has little effect on systemic metabolism of lean mice, suggesting that LSD1 has a context-dependent role in promoting maladaptive changes in obesity. In analysis of insulin target tissues we identified white adipose tissue as the major site of insulin sensitization by GSK-LSD1, where it reduces adipocyte inflammation and lipolysis. We demonstrate that GSK-LSD1 reverses NAFLD in a non-hepatocyte-autonomous manner, suggesting an indirect mechanism potentially via inhibition of adipocyte lipolysis and subsequent effects on lipid partitioning. Pair-feeding experiments further revealed that effects of GSK-LSD1 on hyperglycemia and NAFLD are not a consequence of reduced food intake and weight loss. These findings suggest that targeting LSD1 could be a strategy for treatment of obesity and its associated complications including type 2 diabetes and NAFLD.

T lymphocytes are required for the development of Angiotensin II (Ang II) hypertension in male mice. Cycling female mice are protected against Ang II hypertension, and inducing ovarian failure (menopause) eliminates this protection. We have previously shown that female Rag-1-/- mice with adoptively transferred T cells have significantly reduced renal T cell infiltration compared to males. Therefore, we hypothesized that an increase in renal T cell infiltration underlies the genesis of hypertension in menopausal female mice. 10 week old C57Bl/6 female mice received ip VCD injections for 20 consecutive days to induce ovarian failure. Cyclicity was monitored daily via vaginal cytology. Once in menopause, an osmotic minipump was implanted, releasing Ang II (800ng/kg/min) for 14 days. In a subset of mice, a 17-β estradiol pellet (0.1mg) was implanted subcutaneously. Upon harvest, T cell infiltration was measured via flow cytometry. Baseline blood pressure was similar among all groups. Ang II infusion elicited a significantly greater increase in MAP in menopausal mice compared to cycling control (Ang Δ11±10mmHg vs Meno/Ang Δ32±6, p<0.05), which was prevented by 17-β estradiol replacement (Meno/Ang/E2 Δ5±2mmHg). Despite elevated MAP, there was no significant increase in renal CD3+, CD4+ or CD8+ T lymphocyte infiltration in Meno/Ang group vs control. However, 17-β estradiol replacement decreased IL-2 renal mRNA expression (0.45±0.13 fold), and increased IL-10 expression (2.41±0.73 fold) compared to Ang II-treated menopausal mice. Ang II induced glomerular hypertrophy in all groups, independent of hormonal status (Control 2156±54μm2 vs. Ang 3970±383∗μm2 vs. Meno/Ang 3729±51∗μm2 vs. Meno/E2/Ang 3505±71∗μm2, ∗p<0.05 vs. control). These results demonstrate that the enhanced hypertensive response to Ang II in VCD-treated menopausal mice results from the loss of estrogen function, and does not require an increase in renal T lymphocyte infiltration. Ang II-induced glomerular hypertrophy occurs independently of blood pressure or hormonal status. The VCD model of ovarian failure is a useful model for investigating the underlying mechanisms of estrogen’s role in blood pressure regulation. [ABSTRACT FROM AUTHOR]

There is extensive evidence that activation of the immune system is both necessary and required for the development of Ang II induced hypertension in males and that the central nervous system (CNS) is involved in this response. The purpose of the present study was to determine if there are sex differences in ability of the adaptive immune system to induce Ang II hypertension and the effect of sex on T-cell infiltration into the brain and kidney during Ang II induced hypertension. Rag-1-/- mice, with a genetic deletion of the recombinase-activating gene and lacking both T and B cells were used in this study. Male (n=4) and female (n=4) Rag-1-/- mice both received adoptive transfer of male CD3+ T cells three weeks prior to 14 day Ang II infusion (490 ng/kg/min). Blood pressure was monitored non-invasively via tail cuff. In Rag-1-/- mice, in the absence of T cells, male and female mean arterial pressure (MAP) responses to Ang II were similar, males increased MAP 14.6 mmHg (107.8mmHg to122.5mmHg) and females increased 14.4 mmHg (102.7mmHg to 117.2 mmHg). However, following adoptive transfer of male CD3+ Tcells, Ang II induced a significantly greater increase in blood pressure in males (delta 37.8 mmHg, 107.2mmHg to 139.5mmHg, p<.05) while female mice had similar changes as seen in controls (delta 16.7 mmHg, 110.1mmHg to126.8mmHg). Flow cytometric analysis of CD3+, CD3+CD4+, CD3+CD4+Foxp3 and CD3+CD8+, lymphocyte surface markers from Rag-1-/- males and females was performed on whole blood, splenic, renal and brain tissue. Absolute CD3+ T cell count from spleen suggested that both sexes had equal T cells engraftment. Males had significantly greater numbers of CD3+CD4+, CD3+CD4+Foxp3 and CD3+CD8+ lymphocyte infiltration of the kidney compared to females in both control and AngII infused animals (p<0.05 comparison of males to females). Males also had significantly increased CD3+CD4+ brain infiltration relative to females. These results suggest that intact female Rag-1-/- mice during Ang II infusion are protected from the hypertensive effects of CD3+ male T cell adoptive transfer as compared to their male counterparts and this protection may involve sex differences in T cell infiltration of the kidney and brain. [ABSTRACT FROM AUTHOR]

Premenopausal females are protected from angiotensin II (ANG II)-induced hypertension following the adoptive transfer of T cells from normotensive donors. For the present study, we hypothesized that the transfer of hypertensive T cells (HT) or splenocytes (HS) from hypertensive donors would eliminate premenopausal protection from hypertension. Premenopausal recombination-activating gene-1 (Rag-1) -/- females received either normotensive (NT) or hypertensive cells 3 wk before ANG II infusion (14 days, 490 ng/kg/min). Contrary to our hypothesis, no increase in ANG II-induced blood pressure was observed in the NT/ANG or HT/ANG groups. Flow cytometry demonstrated that renal FoxP3 + T regulatory cells were significantly decreased, and immunohistochemistry showed an increase in renal F4/80 + macrophages in the HT/ANG group, suggesting a shift in the renal inflammatory environment despite no change in blood pressure. Renal mRNA expression of macrophage chemoattractant protein-1 (MCP-1), endothelin-1 (ET-1), and G protein-coupled estrogen receptor-1 (GPER-1) was significantly decreased in the HT/ANG group. The adoptive transfer of hypertensive splenocytes before ANG II infusion (HS/ANG) eliminated premenopausal protection from hypertension and significantly decreased splenic FoxP3 + T regulatory cells compared with females that received normotensive splenocytes (NS/ANG). Expression of macrophage inflammatory protein 1α/chemokine (C-C motif) ligand 3 (MCP-1/CCL3), a potent macrophage chemokine, was elevated in the HS/ANG group; however, no increase in renal macrophage infiltration occurred. Together, these data show that in premenopausal females, T cells from hypertensive donors are not sufficient to induce robust ANG II-mediated hypertension; in contrast, transfer of hypertensive splenocytes (consisting of T/B lymphocytes, dendritic cells, and macrophages) is sufficient. Further work is needed to understand how innate and adaptive immune cells and estrogen signaling coordinate to cause differential hypertensive outcomes in premenopausal females. NEW & NOTEWORTHY Our study is the first to explore the role of hypertensive T cells versus hypertensive splenocytes in premenopausal protection from ANG II-induced hypertension. We show that the hypertensive status of T cell donors does not impact blood pressure in the recipient female. However, splenocytes, when transferred from hypertensive donors, significantly increased premenopausal recipient blood pressure following ANG II infusion, highlighting the importance of further investigation into estrogen signaling and immune cell activation in females.

Although it is known that the prevalence and severity of hypertension increases in women after menopause, the contribution of T cells to this process has not been explored. Although the immune system is both necessary and required for the development of angiotensin II (ANG II) hypertension in men, we have demonstrated that premenopausal women are protected from T cell-mediated hypertension. The goal of the current study was to test the hypotheses that 1 ) female protection against T cell-mediated ANG II hypertension is eliminated following progression into menopause and 2 ) T regulatory cells (Tregs) provide premenopausal protection against ANG II-induced hypertension. Menopause was induced in Rag-1 -/- mice (via 4-vinylcyclohexene diepoxide), and all mice received a 14-day ANG II infusion. Donor CD3 + T cells were adoptively transferred 3 wk before ANG II infusion. In the absence of T cells, systolic blood pressure responses to ANG II were similar to those seen in premenopausal mice (Δ12 mmHg). After adoptive transfer of T cells, ANG II significantly increased systolic blood pressure in postmenopausal females (Δ28 mmHg). A significant increase in F4/80 positive renal macrophages, an increase in renal inflammatory gene expression, along with a reduction in renal expression of mannose receptor C-type 1, a marker for M2 macrophages, accompanied the increase in systolic blood pressure (SBP). Flow cytometric analysis identified that Tregs were significantly decreased in the spleen and kidneys of Rag-1 -/- menopausal mice versus premenopausal females, following ANG II infusion. In a validation study, an anti-CD25 antibody was used to deplete Tregs in premenopausal mice, which induced a significant increase in SBP. These results demonstrate that premenopausal protection against T cell-mediated ANG II hypertension is eliminated once females enter menopause, suggesting that a change in hormonal status upregulates macrophage-induced proinflammatory and T cell-dependent responses. Furthermore, we are the first to report that the presence of Tregs are required to suppress ANG II hypertension in premenopausal females. NEW & NOTEWORTHY Whether progression into menopause eliminated female protection against T cell-mediated hypertension was examined. Menopausal mice without T cells remained protected against angiotensin II (ANG II) hypertension; however, in the presence of T cells, blood pressure responses to ANG II increased significantly in menopause. Underlying mechanisms examined were anti-inflammatory protection provided by T regulatory cells in premenopausal females and renal inflammatory processes involving macrophage infiltration and cytokine activation.

Premenopausal females are resistant to the development of hypertension, and this protection is lost after the onset of menopause, resulting in a sharp increase in disease onset and severity. However, it is unknown how a fluctuating ovarian hormone environment during the transition from perimenopause to menopause impacts the onset of hypertension, and whether interventions during perimenopause prevent disease onset after menopause. A gradual transition to menopause was induced by repeated daily injections of 4-vinylcyclohexene diepoxide (VCD). ANG II (800 ng·kg(-1)·min(-1)) was infused into perimenopausal and menopausal female mice for 14 days. A separate cohort of mice received 17β-estradiol replacement during perimenopause. ANG II infusion produced significantly higher mean arterial pressure (MAP) in menopausal vs. cycling females, and 17β-estradiol replacement prevented this increase. In contrast, MAP was not significantly different when ANG II was infused into perimenopausal and cycling females, suggesting that female resistance to ANG II-induced hypertension is intact during perimenopause. ANG II infusion caused a significant glomerular hypertrophy, and hypertrophy was not impacted by hormonal status. Expression levels of aquaporin-2 (AQP2), a collecting duct protein, have been suggested to reflect blood pressure. AQP2 protein expression was significantly downregulated in the renal cortex of the ANG II-infused menopause group, where blood pressure was increased. AQP2 expression levels were restored to control levels with 17β-estradiol replacement. This study indicates that the changing hormonal environment in the VCD model of menopause impacts the severity of ANG II-induced hypertension. These data highlight the utility of the ovary-intact VCD model of menopause as a clinically relevant model to investigate the physiological mechanisms of hypertension that occur in women during the transition into menopause., (Copyright © 2015 the American Physiological Society.)