Abstract: 【Objective】 In order to repair or restore the function of damaged brain

to regulate signaling pathway by chemicals

which ultimately leads to endogenous neural stem cells (NSC) proliferation and differentiation

has been regarded as an new treatment strategy. Here

we found the effects of nuclear factor erythroid 2-related factor(Nrf2) on C17.2 cell proliferation

and tried to illuminate its possible mechanisms. 【Methods】 Cell viability was tested by MTT assay

and cell proliferation was measured by BrdU incorporation assay. Intracellular ROS formation was monitored by flow cytometry. The protein levels were measured by western blot.【Results】 Two Nrf2 activators

tert-butylhydroquinone(t-BHQ) and sulforaphane(SFN)

promoted the proliferation of C17.2 neural stem cells

which was suppressed by brusatol

an Nrf2 inhibitor. Unexpectedly

intracellular reactive oxygen species (ROS) after treatment with Nrf2 activators or inhibitor were similar to the control group

but Nrf2 activation by t-BHQ or SFN could protect C17.2 cells against hydrogen peroxide (H2O2)-induced damage. Moreover

we observed that microtubule-associated protein 1 light chain 3 (LC3)-II protein level and the green fluorescent protein tagged-LC3 (GFP-LC3) puncta were markedly increased by treatment with t-BHQ or SFN and were significantly decreased by exposure to brusatol in C17.2 cells. Overexpression of Nrf2 not only induced the conversion of LC3-I to LC3-II

but increased C17.2 cell proliferation that was partly reversed by two autophagy inhibitors

ammonium chloride (NH4Cl) and chloroquine (CQ). 【Conclusion】 Our findings suggest that activating autophagy

instead of modulating ROS

plays a pivotal role in the Nrf2-induced C17.2 cell proliferation.