The effect of the sera from severe preeclamptic patients on endothelial cell viability and endothelin-1 in cultured human umbilical vein endothelial cells / 대한산부인과학회잡지

OBJECTIVES: In order to assess the effects of sera from severe preeclamptic patients on endothelial cell viability in vitro and endothelin-1 synthesis in cultured human umbilical vein endothelial cells. METHODS: The cultured human umbilical vein endothelial cells were incubated with media containing 10% sera from women with either preeclamptic patients or normal pregnancies for 24 hours or 48 hours. After then, their viability was measured by colorimetric MTT{3-(4,5-dimethylthiazol-2yl)2,5-diphenyl tetrazolium bromide} assay and their production of endothelin-1 was measured. We also measured the serum levels of endothelin-1 level in sera obtained from the normal and severe preeclamptic pregnancies. RESULTS: The calorimetric MTT assay revealed that after 24 hours, the absorbances in the media treated with normal pregnancies and severe preeclampsia sera were 0.0718+/-0.0078 and 0.0837+/-0.0129, respectively and after 48 hours, they were 0.1133+/-0.0103 and 0.1268+/-0.0186, respectively. Serum obtained from severe preeclampsia did not affect endothelial cell viability. 2. The serum mean levels of endothelin-1 in normal and severe preeclamptic pregnancies were 22.66+/-8.6 fmol/ml and 48.98+/-25.27 fmol/ml. The mean level in preeclamptic sera was significantly higher than that of normal pregnant women. (P<0.05) 3. After 24 hours, the mean amount of endothelin-1 stimulated by normal pregnant and severe preeclamptic sera were 37.52+/-18.41 fmol/ml and 97.58+/-53.64 fmol/ml, respectively. The mean amount of endothelin-1 in preeclamptic sera-treated cells was significantly higher than that of normal pregnant sera-treated cells. (P<0.05). CONCLUSION: The sera from severe preeclamptic women do not affect cell viability but act selectively on specific activation of their function such as endothelin-1 production. And it is necessary that the identification and isolation of the putative serum factor(s) will be performed to resolve the pathogenesis in future.

The effect of steroids on the viability of endothelial cells of stored cornea

The change of endothelial cell viability due to corticosteroid treatment in stored rabbit corneas was investigated. Hydrocortisone was injected into the anterior chamber of enucleated eyeballs which were stored in a moist chamber. After 24,48, or 72 hours of storage, the cornea was removed and stained with trypan blue. The unstained endothelial cells were counted under the light microscope in order to determine the density of viable endothelial cells. The same procedures were done on the contralateral eye with normal saline injected into the anterior chamber instead of hydrocortisone as a control. The density of viable endothelial cells in the corticosteroid-treated group was higher than that of the control group by 1.75%,14.39%, and 27.40% in 24,45, and 72 hour-stored corneas, respectively.