Pancreatic AT1aR Deficiency Decreases Insulin Secretion in Obese C57BL/6 Mice

Robin Shoemaker; Yasir AlSiraj; Jeff Chen; Lisa A Cassis

doi:10.1093/ajh/hpz042

. 2019 Mar 22;32(6):597–604. doi: 10.1093/ajh/hpz042

Pancreatic AT1aR Deficiency Decreases Insulin Secretion in Obese C57BL/6 Mice

Robin Shoemaker ^1,^✉, Yasir AlSiraj ², Jeff Chen ³, Lisa A Cassis ²

PMCID: PMC6508375 PMID: 30903169

Abstract

BACKGROUND

Previously, we demonstrated that obese mice have marked elevations in systemic concentrations of angiotensin II (AngII). Drugs that inhibit the renin–angiotensin system (RAS), including angiotensin type 1 receptor (AT1R) antagonists, have been reported to delay the onset of type 2 diabetes (T2D), suggesting improvements in insulin sensitivity or regulation of pancreatic insulin secretion. Pancreatic islets possess components of the RAS, including AT1R, but it is unclear if AngII acts at islets to regulate insulin secretion during the development of T2D.

METHODS

We deleted AT1aR from pancreatic islets and examined effects on insulin secretion in mice fed a low-fat (LF) or high-fat (HF) diet. In separate studies, to exacerbate the system, we infused HF-fed mice of each genotype with AngII.

RESULTS

Pancreatic AT1aR deficiency impaired glucose tolerance and elevated plasma glucose concentrations in HF, but not LF-fed mice. In HF-fed mice, high glucose increased insulin secretion from islets of AT1aR^fl/fl, but not AT1aR^pdx mice. In AngII-infused mice, following glucose challenge, plasma glucose or insulin concentrations were not significantly different between genotypes. Moreover, high glucose stimulated insulin secretion from islets of AT1aR^fl/fl and AT1aR^pdx mice, presumably related to weight loss, and improved insulin sensitivity in both groups of AngII-infused HF-fed mice.

CONCLUSIONS

Our results suggest that during the adaptive response to insulin resistance from HF feeding, AngII promotes insulin secretion from islets through an AT1aR mechanism. These results suggest the timing of initiation of AT1R blockade may be important in the progression from prediabetes to T2D with β-cell failure.

Keywords: angiotensin, AT1R, blood pressure, diabetes, hypertension, insulin, islets, obesity

Patients with type 2 diabetes (T2D) are at increased risk for cardiovascular disease.¹ Blockade of the renin–angiotensin system (RAS) improves blood pressure control and cardiac outcomes, indicating that overactivity of the RAS contributes to the pathogenesis of cardiovascular diseases. Research investigating mechanisms linking T2D to cardiovascular diseases has identified that drugs inhibiting the RAS may reduce the risk of new-onset diabetes^2–4; however, conflicting results have been reported. In general, clinical trials using RAS inhibitors for cardiovascular conditions demonstrated a reduction in new-onset diabetes, whereas trials where diabetes was the primary outcome demonstrated that RAS inhibition resulted in modest delays in the development of diabetes.^3,5 These findings suggest that the beneficial effects of RAS inhibition to delay the onset of T2D depend on the disease state, where the most effective outcomes are in those patients with an activated RAS.

Angiotensin II (AngII), the main vasoactive component of the RAS, is implicated in the development of insulin resistance⁶ and is reported to regulate insulin secretion from isolated islets, albeit with conflicting findings. AngII inhibited glucose-stimulated insulin secretion in isolated islets,^7,8 while blockade or silencing of AT1R improved insulin secretion.^8–10 In contrast, infusion of AngII enhanced in vivo insulin secretion in mice fed standard murine diet.¹¹ In animal models of T2D, whole-body inhibition of the RAS consistently improves insulin sensitivity and glucose homeostasis.^9,12–14 Taken together, these findings suggest a complex role for AngII in the pathogenesis of diabetes, with possible effects to regulate insulin sensitivity and/or insulin secretion. However, as changes in insulin sensitivity influence pancreatic insulin release, tissue-specific effects of AngII to regulate glucose homeostasis are unclear.

Previously, we demonstrated that C57BL/6 mice with diet-induced obesity are insulin resistant and develop hypertension associated with elevated plasma concentrations of AngII.^15,16 Given that pancreatic islets express components of the RAS, including angiotensin type 1 receptors (AT1R),^7,17–19 we hypothesized that high concentrations of AngII in obese mice act directly at pancreatic AT1aR to regulate insulin secretion. To test this hypothesis, we examined effects of pancreatic AT1aR deficiency on whole-body glucose homeostasis and insulin secretion from islets of low-fat (LF)- and high-fat (HF)-fed mice. Moreover, to exacerbate the system, we infused HF-fed mice with exogenous AngII to define effects of pancreatic AT1aR deficiency on whole-body glucose homeostasis and islet insulin secretion.

MATERIALS AND METHODS

Experimental animals

Studies were conducted according to the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were approved by the University of Kentucky Institutional Animal Care and Use Committee. Female mice with loxP sites flanking exon 3 of the AT1aR gene on a C57BL/6 background (AT1aR^fl/fl) were bred to male AT1aR^fl/fl hemizygous transgenic mice expressing Cre recombinase under the control of the pancreas–duodenum homeobox-1 (pdx-1) promoter to generate mice with pancreas-specific deletion of AT1aR (AT1aR^pdx) or littermate controls (AT1aR^fl/fl). Male mice were used as clinical trials do not indicate sexual dimorphism of T2D.^20,21

Initial studies characterized the efficiency and specificity of pancreatic AT1aR deficiency using 8-week-old male AT1aR^fl/fl and AT1aR^pdx mice (n = 9–13 mice per group), and offspring of male AT1aR^pdx mice bred to female mice carrying the transgene with the ROSA26-stop-lacZ reporter (Jackson Laboratory, Bar Harbor, ME). Male mice, maintained on standard murine diet until 8–10 weeks of age, were randomly assigned to receive either a LF(10% kcal from fat; D12450B; Research Diets, New Brunswick, NJ) or a HF, (60% kcal from fat; D12492; Research Diets) diet fed ad libitum for 16 weeks (n = 4–13 per group). Body weight was quantified weekly.

In a separate study, AT1aR^fl/fl and AT1aR^pdx mice were fed a HF diet (n = 4–7 mice per group) for 12 weeks. At week 13, mice of each genotype were infused via osmotic micropump (Alzet, model 1004) with AngII (1,000 ng/kg/min) for a duration of 4 additional weeks while maintained on the HF diet. During the final week of AngII infusions, plasma glucose and insulin concentrations were quantified. Islets were isolated from anesthetized mice (ketamine/xylazine, 100/10 mg/kg i.p.) of each genotype at study end point (week 4 of AngII infusions, week 17 of HF diet).

Detection of β-galactosidase activity in tissues

Whole organs were fixed in formalin at 4 °C for 1 hour, then rinsed 3 times with buffer (100-nM sodium phosphate, 2-mM MgCl₂, 0.01% sodium deoxycholate, 0.02% NP-40). Organs were incubated overnight in X-gal staining buffer (above rinse buffer with 5-mM potassium ferricyanide, 5-mM potassium ferrocyanide, 1 mg/ml X-gal).

Extraction of DNA and RNA for genotyping and RT-PCR

DNA was extracted from whole pancreas (DNeasy; Quiagen, Alameda, CA) and amplified using the following primers: forward, 5′-TCTTCAAGACTGCTGATGTC, reverse, 5′-GGTTGAGTTGGTCTCAGAC (355-bp product between loxP site 1 and exon 3 of AT1aR gene demonstrates the presence of the floxed AT1aR gene), and 5′-GCAACTATGTCTGTCACTGG (423-bp sequence between loxP sites 1 and 3 demonstrates the deletion of exon 3 of the AT1aR gene). Total RNA was extracted from tissues using the SV total RNA Isolation System (Promega, Madison, WI). Reverse transcription was performed using qScript cDNA SuperMix (Quanta Biosciences, Gaithersburg, MD) and real-time polymerase chain reaction was performed using PerfeCTa SYBR Green FastMix (Quanta Biosciences). Mouse AT1R primers were obtained from Qiagen (QT00261464). Data are expressed as ΔΔCt relative to 18S rRNA (forward: 5′-AGTCGGCATCGTTTATGGTC, reverse, 5′-CGAAAGCATTTGCCAAGAAT).

In vivo glucose-stimulated insulin release and glucose tolerance

For glucose tolerance tests, mice were fasted for 6 hours. Blood glucose concentrations were quantified using a glucometer (Freedom Freestyle Lite; Abbott Laboratories, Abbott Park, IL) immediately before and 15, 30, 60, 90, and 120 minutes following intraperitoneal (i.p.) administration of glucose (1.5 g/kg body weight). Blood samples (10 µl) collected via repeated tail vein stick in conscious mice following a 6 hour fast and again after i.p. administration of glucose (2 g/kg body weight) were used for quantification of plasma glucose concentrations (using a glucometer) and plasma insulin concentrations (quantified by ELISA using a commercial kit; Crystal Chem, Downers Grove, IL). The quantitative insulin sensitivity check index (QUICKI) was used to assess insulin sensitivity with the following formula: 1/[log (fasting insulin, uM/ml) + log (fasting glucose, mg/dl)].¹²

Glucose-stimulated insulin secretion from isolated pancreatic islets

The method for islet isolation has been described previously.²² Briefly, pancreata were perfused in situ through the common bile duct with 5 ml of collagenase (0.6 mg/ml, Collagenase P; Roche, Indianapolis, IN), excised, and incubated in 10 ml of collagenase at 37 ºC for 10 minutes. Islets were purified from exocrine tissue using a sucrose gradient (Histopaque 1077; Sigma-Aldrich, St. Louis, MO), allowed to recover in culture media (RPMI, 10% FBS, 1% penicillin/streptomycin) for 24 hours, and further purified from exocrine tissue by handpicking with a pipette. Purified islets (n = 20 per mouse) were placed in inserts in 12 well plates (Greiner Bio-One, Monroe, NC) and incubated at 37 ºC for 1 hour each in Krebs buffer with 3-mM glucose followed by incubation in Krebs buffer with 25 mM. Insulin concentrations in media and in islets (harvested in HCl–ethanol) were quantified by ELISA and normalized to protein content of islets (BCA assay; ThermoFisher, Rockford, IL).

Statistical analysis

Data are presented as mean ± SEM. All statistical analyses were performed using SigmaPlot, version 12.3. All data passed normality and/or equal variance tests, or logarithmic transformation was used to achieve normality. Two-tailed Student’s t-tests were used for analysis of data between 2 groups. For 2-factor analysis, a 2-way analysis of variance was used to analyze end-point measurements (between group factors of: genotype and time, genotype and diet, treatment and diet, treatment and genotype), followed by Holm-Sidak for post hoc analyses. Glucose tolerance tests were analyzed using a repeated measures 2-way analysis of variance. Significance was denoted at P < 0.05.

RESULTS

Development of a mouse model of pancreatic AT1aR deficiency

Deletion of the AT1aR gene using the Cre–LoxP system driven by the pancreas-specific promoter, pdx-1 (Figure 1a), was confirmed by polymerase chain reaction in DNA extracted from whole pancreas (Figure 1b). AT1aR mRNA abundance was significantly decreased in pancreas from AT1aR^pdx compared to AT1aR^fl/fl mice (Figure 1c; P < 0.05). In contrast, there was no difference in AT1aR mRNA abundance between genotypes in heart, kidney, liver, duodenum, stomach, spleen, lung, soleus, or adipose tissue (Figure 1c). Positive β-galactosidase staining was present in pancreas, duodenum, stomach, hypothalamus and partially in the brain stem and bile duct of AR1aR^pdx mice, and in the stomach and pyloric and hepatopancreatic sphincters in AT1aR^fl/fl mice, consistent with literature indicating endogenous β-galactosidase activity in these tissues^23,24 (Figure 1d). In 8-week-old mice fed standard murine diet, there was no significant difference in body weights or tissue weights between genotypes (heart, liver, kidney, duodenum, stomach, spleen, lung, soleus, adipose; data not shown). However, pancreas weight was significantly reduced in AT1aR^pdx compared to AT1aR^fl/fl mice (0.294 ± 0.011 vs. 0.356 ± 0.14 g, respectively, P < 0.05). However, there was no difference in glucose tolerance (Figure 1e) or in vivo glucose-stimulated insulin secretion (Figure 1f) between genotypes.

Glucose homeostasis is modestly impaired in HF-fed mice with pancreatic AT1aR deficiency

Body weight was significantly increased in HF-fed compared to LF-fed mice (P < 0.001), with no differences in body weight between genotypes (Figure 2a). HF-fed mice developed marked hyperinsulinemia, assessed by quantification of fasting plasma insulin concentrations, compared to LF mice (LF, AT1aR^fl/fl: 2.63 ± 0.22; LF, AT1aR^pdx: 2.91 ± 0.16; HF, AT1aR^fl/fl: 13.92 ± 2.95; HF, AT1aR^pdx: 12.69 ± 1.57 ng/ml; P < 0.05 effect of diet). Chronic HF feeding resulted in significantly increased blood glucose concentrations following an i.p. glucose challenge in both genotypes compared to LF controls (Figure 2b; P < 0.01). However, blood glucose concentrations were significantly elevated at the 15- and 30-minute time points in HF-fed AT1aR^pdx compared to AT1aR^fl/fl mice (Figure 2b; P < 0.05). A separate test was performed to quantify in vivo glucose-stimulated insulin secretion at 15 and 30 minutes following i.p. glucose challenge in HF-fed mice of each genotype. HF-fed AT1aR^pdx mice had significantly elevated plasma glucose concentrations at the 15-minute time point compared to AT1aR^fl/fl mice (Figure 2c; P < 0.05), contributing to an increased AUC (Figure 2d; P < 0.05). Corresponding plasma insulin concentrations were modestly, but not significantly decreased following glucose challenge in HF-fed AT1aR^pdx compared to HF-fed AT1aR^fl/fl mice at each time point (Figure 2e) and when analyzed as area under the curve (Figure 2f; P = 0.07). QUICKI index was used as a surrogate for insulin sensitivity. Although HF-fed mice were less insulin sensitive than LF-fed mice, there was no difference between genotypes (LF, AT1aR^fl/fl: 0.246 ± 0.003; LF, AT1aR^pdx: 0.246 ± 0.003; HF, AT1aR^fl/fl: 0.210 ± 0.005; HF, AT1aR^pdx: 0.209 ± 0.003).

HF feeding increases in vitro insulin secretion from islets isolated from AT1aR^fl/fl, but not AT1aR^pdx mice

To determine if modest reductions in in vivo plasma insulin concentrations of HF-fed AT1aR^pdx mice resulted from reduced insulin release from pancreatic islets, we quantified in vitro insulin secretion under high-glucose (25 mM) conditions from isolated pancreatic islets from mice of each genotype. There was a trend for augmented high glucose-induced insulin secretion from islets of LF-fed AT1aR^pdx compared to LF-fed AT1aR^fl/fl mice (Figure 3a; P = 0.075). In AT1aR^fl/fl mice, islet insulin secretion was increased in HF-fed compared to LF-fed mice (Figure 3a; P < 0.05). In contrast, HF feeding did not increase islet insulin secretion in AT1aR^pdx mice compared to LF controls (Figure 3a). Although insulin content of islets was increased in HF-fed mice compared to LF mice in both genotypes (Figure 3b; P < 0.001), islet insulin content was increased 3-fold in AT1aR^fl/fl mice and 2-fold in AT1aR^pdx mice. To assess the stimulatory response of islets to glucose, islets were incubated under low glucose (3 mM, basal) conditions followed by high-glucose (25 mM, stimulated) conditions, and the stimulatory index (ratio of insulin secreted under stimulated to basal conditions) was calculated. Although glucose-stimulated insulin secretion increased 2- and 3-fold in LF-fed AT1aR^fl/fl and AT1aR^pdx mice, respectively, (Figure 3c), HF feeding markedly reduced the stimulatory effect of glucose on isolated islets, with no differences between genotypes (P < 0.01; Figure 3c.)

Figure 3. — High-fat (HF) feeding increases high glucose-stimulated insulin secretion from islets isolated from *AT1aR*^fl/fl, but not *AT1aR*^pdx mice. (a) Insulin release from pancreatic islets isolated from low-fat (LF)- and HF-fed *AT1aR*^fl/fl and *AT1aR*^pdx mice (n = 20 islets per mouse) incubated in high glucose concentrations (25 mM) for 1 hour. (b) Islet insulin content from LF- and HF-fed *AT1aR*^fl/fl and *AT1aR*^pdx mice. (c) Stimulatory index: ratio of insulin released under high glucose (25 mM) to low glucose (3 mM) concentrations. Data are mean ± SEM from n = 5–11 mice per group. ^*P < 0.05 significance of diet using 2-way analysis of variance.

Pancreatic AT1aR deficiency has no effect on in vivo or in vitro glucose-stimulated insulin secretion in HF-fed mice infused with AngII

C57BL/6 mice of either genotype had robust hyperinsulinemia following 15 weeks of HF feeding associated with hyperglycemia (Figure 2c), suggesting that they had not progressed to β-cell failure. Therefore, we infused AngII to HF-fed mice of each genotype to determine if high exogenous AngII could promote β-cell failure, and whether this would be influenced by pancreatic AT1aR deficiency. Infusion of AngII to HF-fed mice resulted in a significant decrease in body weight (by 24% and 19% in AT1aR^fl/fl and AT1aR^pdx mice, respectively, P < 0.01), with no difference between genotypes at study end point (AT1aR^fl/fl: 38.8 ± 7.8; AT1aR^pdx: 40.9 ± 6.4 g). Plasma glucose and insulin concentrations at baseline and following glucose challenge were decreased with AngII infusion compared to mice of each genotype fed the HF diet for 15 weeks (Figure 4a, b vs. Figure 2c, e). There was no significant difference in glucose or insulin concentrations between genotypes with AngII infusion (Figure 4a, b). Surprisingly, insulin secretion from islets was stimulated by high glucose in of AngII-infused HF-fed mice (Figure 4c; P < 0.05), with no differences between genotypes.

Figure 4. — Glucose-stimulated insulin release in islets isolated from high-fat (HF)-fed mice of each genotype infused with angiotensin II (AngII). *AT1aR*^fl/fl and *AT1aR*^pdx mice were fed a HF diet for 12 weeks, followed by 28 days of infusion with AngII (1,000 ng/kg//min) *via* osmotic pump. (a) Plasma glucose concentrations and (b) corresponding plasma insulin concentrations following a 6 hour fast (0 time point), and 15 and 30 minutes following intraperitoneal (i.p.) glucose administration after 28 days of infusion with AngII and 16 weeks of HF feeding. (c) Insulin release from pancreatic islets (n = 20 islets per mouse) incubated in low (3 mM), followed by high (28 mM) glucose concentrations. Data are mean ± SEM (n = 4–7 mice per group). ^@P < 0.01 overall significance of high glucose using repeated measures 2-way analysis of variance.

DISCUSSION

We defined the role of pancreatic AT1aR on in vivo and in vitro glucose-stimulated insulin secretion in LF- and HF-fed mice, and in HF-fed mice infused with exogenous AngII. Our results demonstrate that, contrary to expectations, pancreatic deficiency of AT1aR modestly impaired in vivo and in vitro glucose-stimulated insulin release in glucose-intolerant obese mice. Elevations in plasma glucose concentrations of HF-fed mice with pancreatic AT1aR deficiency were associated with reduced insulin secretion under high-glucose conditions from pancreatic islets. These results indicate a previously unrecognized role of AngII to increase insulin release through an AT1aR mechanism during the compensatory hyperinsulinemic phase of obesity-induced insulin resistance. Translational relevance of these findings relates to the timing of initiating AT1R blockade to treat cardiovascular conditions associated with T2D, where AT1R inhibition during the early stages of hyperinsulinemia may have negative consequences on insulin release.

As C57BL/6 mice continued to exhibit hyperinsulinemia throughout prolonged HF feeding, we infused mice of each genotype with AngII in an attempt to promote β-cell failure. Infusion of AngII resulted in weight loss in both genotypes associated with improved in vivo and in vitro glucose homeostasis. However, pancreatic AT1aR deficiency had no effect on in vivo or in vitro glucose-stimulated insulin release of AngII-infused mice. These results are consistent with findings in this study from LF-fed mice, where endogenous AngII did not influence glucose homeostasis through pancreatic AT1aR. Taken together these results suggest that during normal physiology (or weight loss in obese mice), pancreatic AT1aR are not primary regulators of glucose-stimulated insulin secretion.

Although effects of AngII to either increase¹¹ or decrease⁷ insulin secretion are conflicting, AngII is consistently implicated in several other mechanisms contributing to overall dysfunction of β-cells, such as inflammation,²⁵ generation of reactive oxygen species,²⁶ and fibrosis.²⁷ Moreover, several studies indicate that AT1R blockade improves glycemia in vivo in models of T2D.^8,12 Previous studies in experimental models where β-cell function was markedly deteriorated, such as islets isolated from db/db mice^8,10 or diabetic rats,^27,28 demonstrated beneficial effects of RAS inhibition on β-cell function. In this study, C57BL/6 mice fed a HF diet for 15 weeks continued to exhibit hyperinsulinemia associated with glucose intolerance. This murine strain has been reported previously to be resistant to effects of high glucose to reduce insulin secretion from islets²⁹ or in vivo insulin responses to glucose challenge,³⁰ even after 10 months of HF feeding.³¹ These data indicate that HF-fed C57BL/6 mice in this study did not progress to β-cell failure, precluding observation of protective effects of pancreatic AT1aR deficiency in an advanced state of T2D.

Factors contributing to β-cell failure include prolonged exposure to metabolic stress, inflammation, endoplasmic reticulum stress, and reactive oxygen species.³² As AngII is implicated in nearly all of these processes, we hypothesized that infusion of exogenous AngII via osmotic pump into HF-fed mice would hasten β-cell dysfunction and decrease insulin secretion, and this effect would be mediated by pancreatic AT1aR. We did not observe a deleterious effect of exogenous AngII infusion on glucose homeostasis in HF-fed mice, potentially related to weight loss-producing effects of AngII infusions.³³ These results suggest that effects of AngII infusion to improve glycemia in vivo and restore the stimulatory response to glucose ex vivo were likely due to peripheral effects of improved insulin sensitivity with weight loss, and not due to direct effects of AngII at the pancreas. Alternatively, effects of increased insulin secretion with pancreatic AT1aR deletion or AngII infusion could be attributed to AngII effects at angiotensin type 2 receptors (AT2R). AT2R are expressed on rodent islets,^34,35 where AngII exerts insulinotropic effects through AT2R activation. Specifically, activation of AT2R by AngII³⁵ or an AT2R agonist³⁶ increased insulin secretion in rats, and administration of an AT2R agonist improved insulin sensitivity in mice.³⁷ Thus, actions of AngII at AT2R may have contributed to improved metabolic parameters with AngII infusion in HF-fed mice of this study.

In summary, deficiency of pancreatic AT1aR reduced in vivo and in vitro glucose-stimulated insulin secretion of HF-fed mice, supporting an initial protective role for AngII at pancreatic AT1aR in the compensatory phase of hyperinsulinemia. In contrast, pancreatic AT1aR deficiency had no effect on in vivo or in vitro glucose-stimulated insulin release in LF-fed mice, or in obese mice losing weight from infusion of exogenous AngII, suggesting that this pathway is not a primary regulator of insulin secretion in normal physiology or in lean states. These results suggest that the timing of initiation of AT1R blockade during the progression from prediabetes to diabetes in obese patients may be important, with potential negative consequences of AT1R blockade at early stages of compensatory hyperinsulinemia.

ACKNOWLEDGMENTS

This study was funded by the National Institutes of Health (NIH) Heart, Lung, and Blood Institute (grant R01HL73085 to L.C.), by the NIH General Medical Sciences (grant P20 GM103527 to L.C.), by the American Heart Association (grant 12PRE12050430 to R.S.), and by the Diabetes Research Center at Washington University (grant 5 P30 DK020579 to L.C.).

DISCLOSURE

The authors declared no conflict of interest.

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26. Hirata AE, Morgan D, Oliveira-Emilio HR, Rocha MS, Carvalho CR, Curi R, Carpinelli AR. Angiotensin II induces superoxide generation via NAD(P)H oxidase activation in isolated rat pancreatic islets. Regul Pept 2009; 153:1–6. [DOI] [PubMed] [Google Scholar]
27. Tikellis C, Wookey PJ, Candido R, Andrikopoulos S, Thomas MC, Cooper ME. Improved islet morphology after blockade of the renin- angiotensin system in the ZDF rat. Diabetes 2004; 53:989–997. [DOI] [PubMed] [Google Scholar]
28. Kaihara M, Nakamura Y, Sugimoto T, Uchida HA, Norii H, Norii H, Hanayama Y, Makino H. Olmesartan and temocapril prevented the development of hyperglycemia and the deterioration of pancreatic islet morphology in Otsuka-Long-Evans-Tokushima Fatty rats. Acta Med Okayama 2009; 63:35–42. [DOI] [PubMed] [Google Scholar]
29. Zraika S, Aston-Mourney K, Laybutt DR, Kebede M, Dunlop ME, Proietto J, Andrikopoulos S. The influence of genetic background on the induction of oxidative stress and impaired insulin secretion in mouse islets. Diabetologia 2006; 49:1254–1263. [DOI] [PubMed] [Google Scholar]
30. Kooptiwut S, Zraika S, Thorburn AW, Dunlop ME, Darwiche R, Kay TW, Proietto J, Andrikopoulos S. Comparison of insulin secretory function in two mouse models with different susceptibility to beta-cell failure. Endocrinology 2002; 143:2085–2092. [DOI] [PubMed] [Google Scholar]
31. Ahrén B, Pacini G. Insufficient islet compensation to insulin resistance vs. reduced glucose effectiveness in glucose-intolerant mice. Am J Physiol Endocrinol Metab 2002; 283:E738–E744. [DOI] [PubMed] [Google Scholar]
32. Prentki M, Nolan CJ. Islet beta cell failure in type 2 diabetes. J Clin Invest 2006; 116:1802–1812. [DOI] [PMC free article] [PubMed] [Google Scholar]
33. Cassis LA, Marshall DE, Fettinger MJ, Rosenbluth B, Lodder RA. Mechanisms contributing to angiotensin II regulation of body weight. Am J Physiol 1998; 274:E867–E876. [DOI] [PubMed] [Google Scholar]
34. Leung KK, Liang J, Zhao S, Chan WY, Leung PS. Angiotensin II type 2 receptor regulates the development of pancreatic endocrine cells in mouse embryos. Dev Dyn 2014; 243:415–427. [DOI] [PubMed] [Google Scholar]
35. Shao C, Zucker IH, Gao L. Angiotensin type 2 receptor in pancreatic islets of adult rats: a novel insulinotropic mediator. Am J Physiol Endocrinol Metab 2013; 305:E1281–E1291. [DOI] [PMC free article] [PubMed] [Google Scholar]
36. Shao C, Yu L, Gao L. Activation of angiotensin type 2 receptors partially ameliorates streptozotocin-induced diabetes in male rats by islet protection. Endocrinology 2014; 155:793–804. [DOI] [PMC free article] [PubMed] [Google Scholar]
37. Quiroga DT, Muñoz MC, Gil C, Pffeifer M, Toblli JE, Steckelings UM, Giani JF, Dominici FP. Chronic administration of the angiotensin type 2 receptor agonist C21 improves insulin sensitivity in C57BL/6 mice. Physiol Rep 2018; 6:e13824. [DOI] [PMC free article] [PubMed] [Google Scholar]

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PERMALINK

Pancreatic AT1aR Deficiency Decreases Insulin Secretion in Obese C57BL/6 Mice

Robin Shoemaker

Yasir AlSiraj

Jeff Chen

Lisa A Cassis