1.Immunoreactivity of Constitutive and Inducible Heat Shock Protein 70 in Human Fetal Retina.
Jeong Hun KIM ; Young Suk YU ; Jin Hyoung KIM ; Yu Jeong KIM ; Jang Won HEO ; Chong Jai KIM
Korean Journal of Ophthalmology 2003;17(1):14-18
The purpose of this study was to measure the level of expression of the inducible heat shock protein70 (Hsp70), the constitutive heat shock protein70 (Hsc70) in the outer nuclear layer and the photoreceptors in the human fetal retina. Fetal eyeballs were selected from fetal autopsy specimens of 12 and 17 to 40 week old fetuses, with no history of congenital anomalies. The retinas had differentiated from neuroblastic cells, into matured sensory retinas, from a gestational age (GA) from 12 to 36 weeks. The Hsp70 and Hsc70 were prominently expressed in the nuclear layers. The Hsc70 was expressed at all GAs and the Hsp70 was expressed from 20 to 33 weeks GA. This period is in accordance with the maturation of the sensory retina. The expression of heat shock protein may be regulated by the development of the fetus, and play an important role in the ocular development.
Embryo and Fetal Development
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Fetus/metabolism
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Heat-Shock Proteins 70/*metabolism
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Human
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Immunohistochemistry
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Retina/*embryology
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Support, Non-U.S. Gov't
2.Effect of the absence of heat shock protein 70.1 (hsp70.1) on retinal photoreceptors in normal and rd mice.
Young Suk YU ; Jun Ho HEO ; Sang Won HWANG ; Jeong Hun KIM ; Jeong Sun SEO ; Sang In KHWARG ; Hum CHUNG
Korean Journal of Ophthalmology 2001;15(2):67-73
The purposes of this study are to elucidate the retinal changes of heat shock protein 70.1 (hsp70.1) knockout mice and to compare them between in normal and in retinal degeneration (rd) mice. Eyes of hsp70.1 wild type (+/+) and knockout (-/-) mice in the C57BL/6 or FVB genetic backgrounds respectively, which were reared in the normal environment, were examined by fundus photography, electroretinography, light microscopy, terminal dUTP nick-end labeling (TUNEL) stain, and immunohistochemistry. In C57BL/6 mice, fundus photography showed no changes between hsp70.1+/+ and -/- mice at 1 and 6 months of age. Electroretinographic examination showed a tendency of decreased amplitude of a- and b-wave with aging in both genotype, but there were not different statistically. The ratios of the thickness of inner nuclear and outer nuclear layer to the retinal thickness were respectively decreased with aging in both genotype, but there were not different statstically. TUNEL assay showed a few positively labeled cells in the ganglion cell, inner nuclear and outer nuclear layers and the immunohistochemistry showed no immunopositivity of hsp70 in the inner segments of photoreceptor cell layer in both genotype. In rd mice, fundus photography showed a narrowing of the retinal vessels at the age of 4 weeks, however, there were no differences of retinal changes including pigment epithelial layer in both genotype. Electroretinographic examination at the postnatal 2, 3 and 4 weeks showed no differences between them. Loss of photoreceptor cell and outer nuclear layers showed no differences in both genotype. In conclusion, there were no differences of the retinal changes at least under the normal environmental condition in hsp70.1+/+ and -/- mice. These results show that hsp70.1-/- mice can be used to study the role of hsp70.1 to the external stress to the retina.
Animal
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Electroretinography
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Fundus Oculi
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Heat-Shock Proteins 70/*deficiency/genetics
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Mice
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Mice, Inbred C57BL
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Mice, Knockout/genetics
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Photoreceptors, Vertebrate/*metabolism
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Protozoan Proteins/genetics
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Reference Values
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Retinal Degeneration/metabolism/*pathology/*physiopathology
3.Protective Effect of Heat Shock Proteins 70.1 and 70.3 on Retinal Photic Injury after Systemic Hyperthermia.
Jin Hyoung KIM ; Jeong Hun KIM ; Young Suk YU ; Seon Mi JEONG ; Kyu Won KIM
Korean Journal of Ophthalmology 2005;19(2):116-121
PURPOSE: This study aimed to determine the relationship between the heat shock protein 70 from hsps70.1 and 70.3 on retinal photic injury after systemic hyperthermia. METHODS: Eight-week-old female C57BL/6 mice were kept at a constant temperature of 41~42 degrees C for 25~30 minutes. After dark-adaptation for 8 hours, intense light of 11000 lux was maintained for 6 hours. Histology and immunohistochemistry for the inducible heat shock protein 70 (hsp70), the constitutive heat shock protein 70 (hsc70), and western blot analysis, reverse transcriptase-polymerase chain reaction for hsp70.1 and hsp70.3 were performed just before photic injury and after 1, 4, 7, and 14 days. RESULTS: Light-induced retinal degeneration was prevented by thermotolerance. After hyperthermia, hsp70 was densely expressed in the inner segment of the photoreceptor layer on the photic injury. Hsp70 expression increased for 4 days after photic injury and slowly decreased thereafter. mRNA from hsp70.3 was induced earlier than that of hsp70.1. CONCLUSIONS: Retinal photic injury was prevented by hyperthermia-induced hsp70. Hsp70 from hsp70.3 may be a rapid and short-lived responder, and that from hsp70.1 is a slower and more sustained responder. Hsp70 from hsp70.3 may be an initial retinal chaperone while hsp70 from hsp70.1 may be a sustained chaperone.
Animals
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Female
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Fever/metabolism/*physiopathology
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Heat-Shock Proteins 70/*metabolism
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In Vitro
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Light/*adverse effects
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Mice
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Mice, Inbred C57BL
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Radiation Injuries/*prevention & control
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Research Support, Non-U.S. Gov't
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Retina/*radiation effects
4.Effects of rosuvastatin in homocysteine induced mouse vascular smooth muscle cell dedifferentiation and endoplasmic reticulum stress and its mechanisms.
Chang-Zuan ZHOU ; Sun-Lei PAN ; Hui LIN ; Li-Ping MENG ; Zheng JI ; Ju-Fang CHI ; Hang-Yuan GUO
Chinese Journal of Applied Physiology 2018;34(1):43-48
OBJECTIVE:
To investigate the effect of rosuvastatin on homocysteine (Hcy) induced mousevascular smooth muscle cells(VSMCs) dedifferentiation and endoplasmic reticulum stress(ERS).
METHODS:
VSMCs were co-cultured with Hcy and different concentration of rosuvastatin (0.1, 1.0 and 10 μmol/L). Cytoskeleton remodeling, VSMCs phenotype markers (smooth muscle actin-α, calponin and osteopontin) and ERS marker mRNAs (Herpud1, XBP1s and GRP78) were detected at predicted time. Tunicamycin was used to induce, respectively 4-phenylbutyrate(4-PBA) inhibition, ERS in VSMCs and cellular migration, proliferation and expression of phenotype proteins were analyzed. Mammalian target of rapamycin(mTOR)-P70S6 kinase (P70S6K) signaling agonist phosphatidic acid and inhibitor rapamycin were used in Rsv treated VSMCs. And then mTOR signaling and ERS associated mRNAs were detected.
RESULTS:
Compared with Hcy group, Hcy+ Rsv group (1.0 and 10 μmol/L) showed enhanced α-SMA and calponin expression (<0.01), suppressed ERS mRNA levels (<0.01) and promoted polarity of cytoskeleton. Compared with Hcy group, Hcy+Rsv group and Hcy+4-PBA group showed suppressed proliferation, migration and enhanced contractile protein expression (<0.01); while tunicamycin could reverse the effect of Rsv on Hcy treated cells. Furthermore, alleviated mTOR-P70S6K phosphorylation and ERS (<0.01)were observed in Hcy+Rsv group and Hcy+rapamycin group, compared with Hcy group; while phosphatidic acid inhibited the effect of Rsv on mTOR signaling activation and ERS mRNA levels (<0.01).
CONCLUSIONS
Rosuvastatin could inhibit Hcy induced VSMCs dedifferentiation suppressing ERS, which might be regulated by mTOR-P70S6K signaling.
Actins
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metabolism
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Animals
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Calcium-Binding Proteins
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metabolism
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Cell Dedifferentiation
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drug effects
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Cells, Cultured
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Endoplasmic Reticulum Stress
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drug effects
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Heat-Shock Proteins
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metabolism
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Homocysteine
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Membrane Proteins
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metabolism
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Mice
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Microfilament Proteins
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metabolism
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Muscle, Smooth, Vascular
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cytology
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Myocytes, Smooth Muscle
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cytology
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drug effects
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Ribosomal Protein S6 Kinases, 70-kDa
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metabolism
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Rosuvastatin Calcium
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pharmacology
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TOR Serine-Threonine Kinases
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metabolism
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X-Box Binding Protein 1
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metabolism
5.Protective Effect of Heat Shock Protein 70 Against Oxidative Stresses in Human Corneal Fibroblasts.
Yun Sang KIM ; Jung Ah HAN ; Tae Bum CHEONG ; Jae Chun RYU ; Jae Chan KIM
Journal of Korean Medical Science 2004;19(4):591-597
We evaluated DNA protection effect of heat shock protein (HSP) against cytotoxic effects of exogenous nitric oxide (NO) and reactive oxygen intermediate (ROI). Cultured human corneal fibroblasts were divided into 4 groups. Control (Group I) was not exposed to a sub-lethal heat treatment. Other 3 groups were exposed to 43 degrees C for 1 hr, then incubated at 37 degrees C during different duration (1, 6, 24 hr, Group II, III, IV, respectively). Expression pattern of HSP 70 was analyzed by Western blot. Cell viability was measured by MTT assay and the relationship between HSP 70 expression and DNA damage was examined by terminal deoxyribonucleotidyl transferase mediated dUTP-digoxigenin nick and labeling (TUNEL) stain and single cell gel electrophoresis. Expression pattern of HSP 70 was dependent on recovery times. Cell viability following heat treatment was significantly increased and the TUNEL positive cell number was decreased at 6 hr. In single cell gel electrophoresis, tail moments were increased in a dose-dependent manner by SNAP and X/XO. Following heat treatment, tail moments showed decreased significantly at 6 hr. These results suggest that induction of HSP 70 by sub-lethal heat treatment is closely related with cytoprotective effects against oxidative stresses in human corneal fibroblasts.
Cell Survival
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Cells, Cultured
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Cornea/*cytology
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DNA Damage
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Dose-Response Relationship, Drug
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Fibroblasts/cytology/drug effects/*metabolism
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Heat
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Heat-Shock Proteins 70/genetics/*metabolism
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Humans
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In Situ Nick-End Labeling
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Nitric Oxide/metabolism
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Nitric Oxide Donors/pharmacology
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*Oxidative Stress
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Reactive Oxygen Species/metabolism
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Research Support, Non-U.S. Gov't
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S-Nitroso-N-Acetylpenicillamine/pharmacology
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Xanthine/pharmacology
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Xanthine Oxidase/pharmacology