Rachel Dahn
Published: 2023
Total Pages: 0
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Preeclampsia (PE) is characterized by endothelial cell (EC) dysfunction, loss of vasodilatory capacity, and loss of EC integrity. In a healthy pregnancy, there is a 'normal' increase in inflammatory factors, including TNF-[alpha]. TNF-[alpha] abundance increases beyond compensatory limits in PE. The first aim examined the requirement of SRC (using inhibitors PP2, PP3, SRC-I1, Dasatinib*, Saracatinib*, *denotes clinical inhibitor), MEK (using inhibitors U0126, PD98059, PD0325901), and P38MAPK (using inhibitors SB203580, BIRB0796) in maintaining resistance (a hallmark of EC integrity) in P-UAECs via ECIS in the absence and presence of TNF-[alpha]. Our results showed that these inhibitors (and combinations) have differential responses in the absence and presence of TNF-[alpha]. While some inhibitors (and combinations) were stimulatory in the absence, none were able to provide complete or full protection in the presence of TNF-[alpha]-mediated destruction. We concluded the resistance of the P-UAEC monolayer is regulated by SRC, MEK, and P38MAPK. The second aim was to identify the effect of other inflammatory mediators, including interleukin 1[beta] (IL-1[beta]), interleukin 6 (IL-6), and interleukin 8 (IL-8) in the absence and presence of TNF-[alpha] (using P-UAECs via ECIS). In addition, we attempted to discern the effects of GP130 and the downstream signaling pathway JAK (using inhibitor AG490) and SRC (using inhibitor PP2) on EC integrity in the absence and presence of TNF-[alpha], IL-1[beta], or TNF+IL-1[beta]. In the absence of TNF-[alpha] several inhibitors (or combinations) increased resistances above control. However, none of the agents tested (alone or in combination) could prevent or provide full rescue of P-UAEC integrity from the effects of TNF-[alpha] alone or the IL-1[beta]+TNF-[alpha] combination. We conclude that while IL-1[beta] and TNF-[alpha] share some signaling pathways, they are clearly under distinct mechanisms of control. In the third and final aim, we examined if P-UAECs undergo cell surface changes (a process known as immune modulation) in response to long-term exposure (20hrs) to different cytokines TNF-[alpha] , IL-1[beta], or IFN-[gamma] and identify if there were subpopulations of cells (using multiparameter flow cytometry experiments). The novel discovery of this collection of work is that PUAEC subpopulations were formed and changes of these populations depend upon the type of cytokine treatment. Immune modulation is indeed at play in regulating endothelial subpopulations, damage, and, therefore, the pathogenesis of preeclampsia.