Determination of conventional protein kinase C isoforms involved in high intraocular pressure-induced retinal ischemic preconditioning of rats

Ding, Jingwen; Ding, Ning; Wang, Ningli; Lu, Qingjun; Lu, Ningning; Yang, Diya; Bu, Xiangning; Han, Song; Li, Junfa

doi:10.1016/j.visres.2008.10.018

Cited by 7 publications

(5 citation statements)

References 32 publications

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“…To determine changes in PKC isoform-specific expression in the ventricles of rats following chronic hypoxia-induced PH, immunostaining with cPKCα, βI, βII and γ, and nPKCδ was carried out as reported ( 26 ). LV and RV tissues were fixed in 10% formaldehyde in PBS at 4°C for 24 h. The tissues were dehydrated through graded alcohol, cleared with xylene, embedded in paraffin, and sectioned at a thickness of 5 µm.…”

Section: Methodsmentioning

confidence: 99%

Protein kinase C isozyme expression in right ventricular hypertrophy induced by pulmonary hypertension in chronically hypoxic rats

Zeng

Liang

Jiang

et al. 2017

Molecular Medicine Reports

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In chronic hypoxia, pulmonary hypertension (PH) induces right ventricular hypertrophy (RVH). Evidence indicates that protein kinase C (PKC) serves a crucial role in hypoxia-induced RVH. The present study investigated PKC isoform-specific expression and its involvement in RVH. Rats were exposed to normobaric hypoxia for a number of days to induce PH. PKC isoform-specific membrane translocation and protein expression in the myocardium were evaluated by western blotting and immunostaining. A total of six isoforms of conventional PKC (cPKC; α, βI and βII) and of novel PKC (nPKC; δ, ε and η), were detected in the rat myocardium. Hypoxic exposure (1–21 days) induced PH with RVH and vascular remodeling. nPKCδ membrane translocation at 3–7 days and cPKCβI expression at 1–21 days in the RV following hypoxic exposure were significantly decreased as compared with the normoxia control group. Membrane translocation of cPKCβII at 14–21 days and of nPKCη at 7–21 days in the left ventricle following hypoxic exposure was significantly increased when compared with the control. The results of the present study suggested that the alterations in membrane translocation, and nPKCδ and cPKCβI expression, are associated with RVH following PH, and the upregulation of cPKCβII membrane translocation is involved in left-sided heart failure.

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Section: Methodsmentioning

confidence: 99%

Protein kinase C isozyme expression in right ventricular hypertrophy induced by pulmonary hypertension in chronically hypoxic rats

Zeng

Liang

Jiang

et al. 2017

Molecular Medicine Reports

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“…As described previously [26,27], equal amounts of vascular protein (50 μg from either cytosolic or particulate fractions, and 20 μg from whole tissue homogenate) per lane were loaded on 10% SDS-PAGE gels. After separation, the proteins were transferred onto polyvinylidene difluoride (PVDF) membrane (GE Healthcare, Buckinghamshire, UK) at 4°C.…”

Section: Methodsmentioning

confidence: 99%

“…To determine the changes of PKC isoform-specific expressions in pulmonary arteries of rats after chronic hypoxia-induced pulmonary hypertension, immunostaining with cPKCα, βI, βII, γ and nPKCδ (Santa Cruz Biotechnology Inc., CA, USA) were performed as reported [27]. Lungs were infused by the fixative (3% formaldehyde in PBS) via the trachea, and then immersed in the fixative at 4°C for 24 h. The tissues were dehydrated, embedded in paraffin, and sectioned at a thickness of 5 μm.…”

Section: Methodsmentioning

confidence: 99%

Determination of PKC isoform-specific protein expression in pulmonary arteries of rats with chronic hypoxia-induced pulmonary hypertension

et al. 2012

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SummaryBackgroundEvidence indicates that protein kinase C (PKC) plays a pivotal role in hypoxia-induced pulmonary hypertension (PH), but PKC isoform-specific protein expression in pulmonary arteries and their involvement in hypoxia-induced PH are unclear.Material/MethodsMale SD rats (200–250 g) were exposed to normobaric hypoxia (10% oxygen) for 1, 3, 7, 14 and 21 d (days) to induce PH. PKC isoform-specific membrane translocation and protein expression in pulmonary arteries were determined by using Western blot and immunostaining.ResultsWe found that only 6 isoforms of conventional PKC (cPKC) α, βI and βII, and novel PKC (nPKC) δ, ɛ and η were detected in pulmonary arteries of rats by Western blot. Hypoxic exposure (1–21 d) could induce rat PH with right ventricle (RV) hypertrophy and vascular remodeling. The cPKCβII membrane translocation at 3–7 d and protein levels of cPKCα at 3–14 d, βI and βII at 1–21 d decreased, while the nPKCδ membrane translocation at 3–21 d and protein levels at 3–14 d after hypoxic exposure in pulmonary arteries increased significantly when compared with that of the normoxia control group (p<0.05 vs. 0 d, n=6 per group). In addition, the down-regulation of cPKCα, βI and βII, and up-regulation of nPKCδ protein expressions at 14 d after hypoxia were further confirmed by immunostaining.ConclusionsThis study is the first systematic analysis of PKC isoform-specific membrane translocation and protein expression in pulmonary arteries, suggesting that the changes in membrane translocation and protein expression of cPKCα, βI, βII and nPKCδ are involved in the development of hypoxia-induced rat PH.

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“…2.6 | Analysis of IL-17R membrane translocation According to our previous reports, 19,20 the frozen samples were thawed and homogenized in buffer A (50 mM Tris-Cl, pH 7.5, containing 2 mM EDTA, 1 mM EGTA, 100 mM iodoacetamide [SH-group blocker], 5 μg/mL each of leupeptin, aprotinin, pepstatin A and chymostatin, 50 mM potassium fluoride, 50 nM okadaic acid, 5 mM sodium pyrophosphate). Homogenates were centrifuged at 100 000g for 30 minutes at 4°C to yield the supernatants as cytosolic fraction.…”

Section: Reverse Transcription Quantitative Real-time Polymerase Chai...mentioning

confidence: 99%

Interleukin‐17A‐mediated alleviation of cortical astrocyte ischemic injuries affected the neurological outcome of mice with ischemic stroke

Dai

Liu

et al. 2019

J of Cellular Biochemistry

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We previously reported that astrocytes are the main sources of interleukin (IL)-17A production that could aggravate neuronal injuries in ischemic stroke. However, the effects of IL-17A on ischemic astrocytes themselves and the underlying molecular mechanism are still unclear. In this study, we found that recombinant mouse (rm) IL-17A could significantly (P < 0.05 or <0.001) alleviate 1-hour oxygen-glucose deprivation (OGD)/reoxygenation (R) 24-hour-induced ischemic injuries in cortical astrocytes with a dose-dependent manner (n = 6 per group). The Western blot and cell cycle analysis results revealed that rmIL-17A significantly (P < 0.05) inhibited procaspase-3 cleavage without affecting cell proliferation in 1-hour OGD/R 24-hour-treated cortical astrocytes (n = 6 per group). Among the five IL-17 receptor subunits (IL-RA, -RB, -RC, -RD, and -RE), only IL-17RA (P < 0.01) and -17RC (P < 0.05) membrane translocation (not messenger RNA and protein) levels were downregulated in cortical astrocytes following 1-hour OGD/reperfusion 24 hours, and rmIL-17A could significantly (P < 0.05 or <0.001) inhibit this downregulation (n = 6 per group). To further verify the impact of IL-17A on the neurological outcome of ischemic stroke, we found that the intracerebroventricular injection of IL-17A neutralizing monoclonal antibody remarkably (P < 0.001) reduced the astrocyte activation and improve neurological function (P < 0.05 or <0.01) of mice following 1-hour middle cerebral artery occlusion/reperfusion (R) 3 to 7 days (n = 6 or 8 per group). These results suggested that IL-17A-mediated alleviation of cortical astrocyte ischemic injuries could affect the neurological outcome of mice with ischemic stroke, which might be mainly dependent on the cell apoptosis pathway through inhibiting the downregulation of IL-17RA and -17RC membrane translocations. K E Y W O R D S astrocytes, interleukin-17A, interleukin-17 receptor subunits, middle cerebral artery occlusion, oxygen-glucose deprivation

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Determination of conventional protein kinase C isoforms involved in high intraocular pressure-induced retinal ischemic preconditioning of rats

Cited by 7 publications

References 32 publications

Protein kinase C isozyme expression in right ventricular hypertrophy induced by pulmonary hypertension in chronically hypoxic rats

Protein kinase C isozyme expression in right ventricular hypertrophy induced by pulmonary hypertension in chronically hypoxic rats

Determination of PKC isoform-specific protein expression in pulmonary arteries of rats with chronic hypoxia-induced pulmonary hypertension

Interleukin‐17A‐mediated alleviation of cortical astrocyte ischemic injuries affected the neurological outcome of mice with ischemic stroke

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