Objective      To determine the role of sonic hedgehog (Shh) signaling in the differentiation of rat bone marrow stromal cells (MSCs) into neuron-like cells induced by resveratrol in vitro.       Methods      The rat MSCs were isolated and cultured by whole bone marrow adherent culture

and then induced to differentiate into neuron-like cells with serum-free DMEM/F12 medium containing resveratrol. Cell morphology was observed with invert microscopy. Indirect immunofluorescence assay and Western blotting were applied to detect the protein expression of NSE

MAP-2

Smo and Gli1

and the translocation of Smo and Gli1.       Results      Almost all MSCs and the control cells were poor positive for NSE. The differentiated cells showed neuron-like morphology with the cell body shrinkage and extend long processes after by resveratrol induction

and were positive to NSE and MAP-2 but negative to GFAP. Immunofluorescence assay confirmed that MSCs had primary cilia

and expressed Smo and Gli1 in the cytoplasm. After resveratrol induction

Smo entered into the primary cilia from the cytoplasm while Gli1 translocated to the nucleus from the cytoplasm. Meanwhile

Western blot analysis indicated that the expression of Smo and Gli1 protein was gradually enhanced with the prolongation of induction (P0.01). However

the control cells had no such marked changes.       Conclusion      Resveratrol induces MSCs to differentiate into neuron-like cells. Moreover

Shh signaling pathway is activated during induced process

suggesting that Shh signaling may play an important role in the differentiation.