TY - JOUR
T1 - Heat shock protein-70 and 4-hydroxy-2-nonenal adducts in human placental villous tissue of normotensive, preeclamptic and intrauterine growth restricted pregnancies
AU - Hnat, Michael D.
AU - Meadows, Juliana W.
AU - Brockman, Diane E.
AU - Pitzer, Brad
AU - Lyall, Fiona
AU - Myatt, Leslie
PY - 2005/9
Y1 - 2005/9
N2 - Objective: The purpose of this study was to compare immunohistochemical expression of heat shock protein-70 (hsp70), a marker for oxidative stress, and 4-hydroxy-2-nonenal adducts (HNE), a marker for lipid peroxidation, in placental villous tissue of normotensive, preeclampsia, and intrauterine growth restricted (IUGR) pregnancies. Study design: Placentas were collected and flash frozen in liquid nitrogen after delivery from normotensive pregnancies (n = 5), and pregnancies complicated by preeclampsia (n = 5), IUGR (n = 5), and preeclampsia plus IUGR (n = 4). Cryosections were cut and immunostained with polyclonal anti-hsp70 and monoclonal anti-HNE antibodies using Vectastain Elite ABC kit. Normal rabbit serum or mouse IgG were used as negative controls. Three independent observers, blinded to identity of tissue, examined each slide to identify cellular localization and intensity of the immunostaining. Western blot analysis and scanning densitometry were used to quantify and compare the amount of hsp70 and HNE adducts present in tissue homogenates. Results: Positive immunostaining for both antibodies was observed in cytoplasm of syncytiotrophoblasts, extravillous trophoblasts, vascular smooth muscle, and endothelial cells for all groups. Expression of hsp70 and HNE adducts was reported as observers' mean stained intensity. Overall, kappa showed good agreement between observers. Immunostaining intensity was similar in all tissue types for each group with the exception that immunostaining was significantly more intense in the vascular endothelium of the preeclamptic group for HNE adducts (P = .02) and significantly less intense in the IUGR group for hsp70 (P = .013). Scanning densitometric analysis of the Western blots showed no significant difference in total hsp70 and HNE adducts expression in all 4 tissue groups. Conclusion: Immunohistochemistry showed local changes for oxidative stress and lipid peroxidation in the vascular endothelium from placentas of preeclamptic and IUGR pregnancies. However, these changes were masked when studying tissue homogenates.
AB - Objective: The purpose of this study was to compare immunohistochemical expression of heat shock protein-70 (hsp70), a marker for oxidative stress, and 4-hydroxy-2-nonenal adducts (HNE), a marker for lipid peroxidation, in placental villous tissue of normotensive, preeclampsia, and intrauterine growth restricted (IUGR) pregnancies. Study design: Placentas were collected and flash frozen in liquid nitrogen after delivery from normotensive pregnancies (n = 5), and pregnancies complicated by preeclampsia (n = 5), IUGR (n = 5), and preeclampsia plus IUGR (n = 4). Cryosections were cut and immunostained with polyclonal anti-hsp70 and monoclonal anti-HNE antibodies using Vectastain Elite ABC kit. Normal rabbit serum or mouse IgG were used as negative controls. Three independent observers, blinded to identity of tissue, examined each slide to identify cellular localization and intensity of the immunostaining. Western blot analysis and scanning densitometry were used to quantify and compare the amount of hsp70 and HNE adducts present in tissue homogenates. Results: Positive immunostaining for both antibodies was observed in cytoplasm of syncytiotrophoblasts, extravillous trophoblasts, vascular smooth muscle, and endothelial cells for all groups. Expression of hsp70 and HNE adducts was reported as observers' mean stained intensity. Overall, kappa showed good agreement between observers. Immunostaining intensity was similar in all tissue types for each group with the exception that immunostaining was significantly more intense in the vascular endothelium of the preeclamptic group for HNE adducts (P = .02) and significantly less intense in the IUGR group for hsp70 (P = .013). Scanning densitometric analysis of the Western blots showed no significant difference in total hsp70 and HNE adducts expression in all 4 tissue groups. Conclusion: Immunohistochemistry showed local changes for oxidative stress and lipid peroxidation in the vascular endothelium from placentas of preeclamptic and IUGR pregnancies. However, these changes were masked when studying tissue homogenates.
KW - Heat shock protein (hsp)
KW - Hydroxynonenal (HNE)
KW - Intrauterine growth restriction
KW - Lipid peroxidation
KW - Oxidative stress
KW - Placenta
KW - Preeclampsia
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U2 - 10.1016/j.ajog.2005.01.059
DO - 10.1016/j.ajog.2005.01.059
M3 - Article
C2 - 16150283
AN - SCOPUS:24344445304
SN - 0002-9378
VL - 193
SP - 836
EP - 840
JO - American journal of obstetrics and gynecology
JF - American journal of obstetrics and gynecology
IS - 3
ER -