Biochanin A Reduces Drug-Induced p75NTR Expression and Enhances Cell Survival: A New In Vitro Assay for Screening Inhibitors of p75NTR Expression

Abstract

Following spinal cord injury (SCI) or peripheral neuropathy, increased levels of the p75^NTR death receptor initiate the signal transduction cascade leading to cell death. Investigations of compounds that may ameliorate neuronal cell death have largely used rodent models, which are time consuming, expensive, and cumbersome to perform. Previous studies had demonstrated that steroids, particularly dexamethasone and its analog methylprednisolone sodium succinate, exhibit limited neuroprotective effects against neuronal injury. Significantly, many naturally occurring nonsteroidal plant compounds exhibit structural overlap with steroids. In this report, we present an in vitro cellular screen model to practically examine the efficacy of various phytoestrogens in modulating the ibuprofen-induced expression of p75^NTR and reduced cell survival of CCFSTTG1 and U87MG cells in a rescue (postinjury) or prevention (preinjury) regimen. We show that the phytoestrogen, biochanin A, and, to a lesser extent, genistein are more effective than dexamethasone at reducing p75^NTR expression and improving the viability of U87MG and CCFSTTG1 before and after p75^NTR induction. Furthermore, these studies implicate biochanin A's inactivation of p38-MAPK as a possible contributor to reducing p75^NTR with associated increased cell survival. This new in vitro assay facilitates a more time-efficient screening of compounds to suppress p75^NTR expression and increase neuronal cell viability prior to their evaluation in animal models of neurological diseases.