Animal procedures were approved by the University of Melbourne Ethics Committee (#1613867) and adhered to the National Health and Medical Research Council of Australia guidelines and the Guide for the Care and Use of Laboratory Animals (76 ). To explore the role of microglia in retinal vasomodulation, Cx3cr1GFP/+ and Cx3cr1GFP/GFP mice were used which have one or both alleles of the monocyte-specific receptor, Cx3cr1, replaced with EGFP (77 (link)). To show that Cx3cr1 labels microglia within healthy retina and not infiltrating monocytes, immunohistochemistry was performed with select markers (SI Appendix, Fig. S1 ). NG2-DsRed pericyte reporter mice were used to explore pericyte-microglial contact and were provided by B.T.S. Hyperglycemia was induced in male adult (6- to 8-wk-old) dark agouti rats via a single intraperitoneal injection of STZ (55 mg/kg, in trisodium citrate buffer, pH 4.5; Sigma-Aldrich), with control animals receiving an equivalent volume of vehicle. A separate cohort of animals was treated with candesartan cilexetil (10 µg/mL; Sigma-Aldrich, #SML0245) or vehicle (PEG400/Ethanol/Kolliphor EL/water, 10:5:2:83; Sigma-Aldrich) in their drinking water, 24 h after diabetes induction. In vivo imaging and tissue isolation was performed after 4 wk of diabetes.
Candesartan cilexetil
Manufactured by Merck Group
Sourced in United States
Candesartan cilexetil is a laboratory product manufactured by Merck Group. It is a medication used to treat high blood pressure. The product's core function is to act as an angiotensin II receptor blocker, which helps to lower blood pressure by relaxing blood vessels.
Automatically generated - may contain errors
Lab products found in correlation
Valsartan, by Merck Group (2 mentions)
Trisodium citrate buffer, by Merck Group (1 mentions)
Telmisartan, by Cayman Chemical (1 mentions)
Eprosartan, by Merck Group (1 mentions)
Candesartan, by Cayman Chemical (1 mentions)
Docusate sodium, by Merck Group (1 mentions)
Sodium lauryl sulfate, by Merck Group (1 mentions)
Domiphen bromide, by Merck Group (1 mentions)
Sodium dodecylbenzenesulfonate, by Merck Group (1 mentions)
Cetylpyridinium bromide, by Merck Group (1 mentions)
Oxldl, by Thermo Fisher Scientific (1 mentions)
Mdivi 1, by Merck Group (1 mentions)
Smooth muscle basal medium 2, by Lonza (1 mentions)
Smc growth medium, by Lonza (1 mentions)
Dabrafenib, by Selleck Chemicals (1 mentions)
Pd184352, by Selleck Chemicals (1 mentions)
Sch772984, by Selleck Chemicals (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
Potassium chloride (kcl), by Merck Group (1 mentions)
5 protocols using candesartan cilexetil
1
Microglia Role in Retinal Vasomodulation
2
Angiotensin II Receptor Antagonist Protocols
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Azilsartan (SML-0432. Sigma–Aldrich), candesartan (9003239, Cayman), eprosartan (E2535, Sigma–Aldrich), fimasartan (SML3256, Sigma–Aldrich Cayman), telmisartan (11615, Cayman), and valsartan (SML0142, Sigma–Aldrich) were dissolved in DMSO and used in in vitro studies. For in vivo studies, we used candesartan cilexetil (SML0245, Sigma–Aldrich) dissolved in PBS.
3
Screening Bacterial Toxins and Drug Library
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All bacterial toxins were purchased from List Biological Laboratories (Campbell, CA, USA). An FDA-approved drug library comprising of 1,585 drugs was purchased from Johns Hopkins University Bloomberg School of Public Health, titled, Johns Hopkins Clinical Compound Library (JHCCL). The drugs in the library were kept at −20 °C as 3.3 mM stock solutions in sealed microtiter plates and were made using DMSO as solvent. Compounds of interest were repurchased and reproduced from 3.3 mM solutions. Salmon sperm protamine, cetylpyridinium bromide, domiphen bromide, cetalkonium chloride, sodium lauryl sulfate, ascorbic acid 6-palmitate, sodium dodecylbenzenesulfonate, docusate sodium, and candesartan cilexetil were purchased from Sigma-Aldrich (St. Louis, MO, USA). Thyropropic acid was purchased from Toronto Research Chemicals Inc (Toronto, Ontario, Canada). All drugs were prepared at 3.3 mM using DMSO as a solvent. Full-length human and truncated salmon and human protamines were synthesized by LifeTein (Hillsborough, NJ, USA). All chemicals and peptides in this study had purities greater than 98%.
4
Ox-LDL-Induced Senescence in Aortic SMCs
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Human aortic smooth muscle cells were purchased from LONZA (Portsmouth, NH, USA) and cultured in smooth muscle basal medium-2 (Lonza, Cat #CC-3181) supplemented with SMC growth medium (Lonza, Cat #CC-4149), as described previously (18 (link)). Prior to stimulation, the cells were starved in 0.5 % FBS-containing medium for 16 h. The cells were treated with 20 μmol/L ox-LDL (Thermo Fisher Scientific, Cat #L34357). Time of the exposure to ox-LDL was 24 h for assessment of senescence in immunoblot (detections of p53 and p21) and in Senescence-associated β-galactosidase (SA-β gal) staining, 12 h for mitochondrial autophagy in immunohistochemistry, and 3 h for other experiments. The following inhibitors were used: 20 μmol/L Mdivi-1 (Sigma, Cat #M0199), Drp1 inhibitor; 10 μmol/L candesartan cilexetil (Candesartan) (Sigma-Aldrich, Cat #SML0245), ARB; 10 μmol/L Dabrafenib (Selleck, Cat #S2807), RAF inhibitor; 2 μmol/L PD184352 (Selleck, Cat #S1020), MEK inhibitor; 2 μmol/L SCH772984 (Selleck, Cat #S7101), ERK inhibitor. For the inhibition experiments, cells were treated with each inhibitor for 1 h following the addition of ox-LDL. Control cells were exposed to the same amount of dimethyl sulfoxide (DMSO). The dose of Candesartan was based on the previous report (19 (link)).
5
Quantification of L-Kynurenine and Metabolites
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L-Kynurenine (sulfate salt), candesartan cilexetil, irbesartan, losartan potassium, olmesartan (RNH-6270), telmisartan, valsartan, dimethyl sulfoxide (DMSO), sodium chloride, potassium chloride, magnesium sulfate, calcium chloride, sodium phosphate monobasic, sodium phosphate dibasic, glucose, distilled water, Trizma base, acetic acid, pyridoxal 5′-phosphate, 2-mercaptoethanol, pyruvate, and glutamine were purchased from Sigma-Aldrich. Substances used for highperformance liquid chromatography (HPLC) were obtained from J.T. Baker Chemicals and from Sigma-Aldrich.
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