Extracellular ATP induces albuminuria in pregnant rats.

BACKGROUND: As circulating plasma ATP concentrations are increased in pre-eclampsia, we tested whether increased plasma ATP is able to induce albuminuria during pregnancy. METHODS: Pregnant (day 14) and non-pregnant rats were infused with ATP (3000 microg/kg bw) via a permanent jugular vein cannula. Albuminuria was determined, and blood samples were taken for leukocyte counts, plasma ATP and plasma haemopexin activity. At Day 20 of pregnancy, rats were sacrificed, fetuses and placentas weighed and kidney and placental tissue were snap frozen for immunohistology. RESULTS: ATP infusion induced albuminuria exclusively in pregnant rats, together with increased neutrophil counts, decreased staining for glomerular sialoglycoproteins and CD39 expression, significant intraglomerular monocyte infiltration and increased glomerular intracellular adhesion molecule-1 (ICAM-1) expression. Plasma haemopexin activity was increased in saline-infused pregnant rats as compared to non-pregnant rats but was inhibited in pregnant ATP-infused rats (to non-pregnant levels). At the end of pregnancy (Day 20), increased plasma ATP level was exclusively seen in ATP-infused pregnant rats. In pregnant rats as compared with non-pregnant rats, we found decreased expression of glomerular AT-1 receptors, which was increased after ATP infusion exclusively in pregnant animals. CONCLUSION: The present study shows that ATP infusion induced a pro-inflammatory response leading to glomerular albuminuria exclusively in the pregnant rat. Why extracellular ATP showed this pro-inflammatory response exclusively in the pregnant condition is unclear but is probably related with relatively enhanced non-specific immunity and inflammatory reactions characteristic for the pregnant condition.

Species differences in the effect of pregnancy on lymphocyte cytokine production between human and rat.

In the present study, we evaluated whether lymphocyte cytokine production during human and rat pregnancy shifts toward T helper cell type 2 (Th2) cytokine production. Therefore, blood samples were taken during the follicular and luteal phase and during pregnancy in rats and humans. Whole blood was ex vivo-stimulated with phorbol 12-myristate 13-acetate and calcium ionophore and intracellular interferon-gamma (IFN-gamma) and interleukin (IL)-4 production, and the percentage of cells in the various lymphocyte populations was measured using flow cytometry. Rats and humans adapted their immune responses to pregnancy but have different strategies: During human pregnancy, the percentage of lymphocytes producing IFN-gamma was decreased, and the percentage IL-4-producing lymphocytes was not affected. The rat adapts its immune response to pregnancy by decreasing the total number of the various lymphocyte populations, and the percentage of IFN-gamma- or IL-4-producing lymphocytes was not affected or increased (% IFN-gamma-producing cytotoxic lymphocytes). It is speculated that during rat pregnancy, there is no need to decrease the number of IFN-gamma-producing lymphocytes, as in nonpregnant rats, the total number of IFN-gamma-producing lymphocytes after stimulation is relatively low, and there is no necessity for a further decrease. In nonpregnant humans, the percentage IFN-gamma-producing lymphocytes is much higher and probably dangerous for pregnancy, and therefore, this percentage needs to decrease during pregnancy. In conclusion, although the data from humans concur with the Th1/Th2 paradigm, the data from rats do not concur with this paradigm. The present studies therefore challenge the classical Th1/Th2 paradigm during pregnancy.

Altered monocyte function in experimental preeclampsia in the rat.

OBJECTIVES: In the present study, we evaluated functional activity of monocytes in experimental preeclampsia induced by low-dose endotoxin infusion. STUDY DESIGN: Pregnant (n = 12) and cyclic rats (n = 12) were equipped with a permanent jugular vein cannula and infused with either low-dose endotoxin or saline. One day before the infusion, and 4, 24, 72, and 168 hours after the infusion, blood samples (400 microL) were taken and white blood cell (WBC) and differential cell counts were measured. Samples were (re)stimulated with endotoxin, and the percentage of tumor necrosis factor-alpha (TNFalpha) producing monocytes was measured. RESULTS: During experimental preeclampsia, monocyte TNFalpha production is persistently decreased, while total WBC and granulocyte counts are persistently increased compared with normal pregnant rats. No persistent effect of endotoxin was found in cyclic rats. CONCLUSION: Because decreased endotoxin-induced TNFalpha production is a feature of activated monocytes, the present results indicate that monocytes are persistently activated in experimental preeclampsia. Increased WBC counts and granulocyte numbers in these rats also point to an activated inflammatory response.

Developing ovarian follicles inhibit the endotoxin-induced glomerular inflammatory reaction in pseudopregnant rats.

PROBLEM: We tested the hypothesis that developing ovarian follicles produce factors inhibiting the endotoxin induced inflammatory response. METHOD OF STUDY: Pseudopregnant rats were treated with FSH to induced follicular development (FSH-rats). For control we used untreated pseudopregnant rats (PSP-rats) and rats in the follicular phase of the ovarian cycle (C-rats). All rats were infused with either saline or endotoxin. Three days after the infusion rats were sacrificed and kidney specimens were snapfrozen. Cryostat kidney sections were stained for the presence of monocytes, granulocytes, CD11a- and CD11b-positive cells and for ICAM-1 expression. RESULTS: The results show that induction of follicular development in pseudopregnant rats inhibited glomerular infiltration of monocytes and CD11b(+) cells, while it did not affect the other parameters, i.e. glomerular granulocyte number, CD11a(+) cells and glomerular ICAM-1 expression. CONCLUSION: Developing ovarian follicles produce factors inhibiting monocyte responses to endotoxin.