〔Abstract〕 Objective To investigate the effect of hydrogen sulfide(H2S) on reactive proliferation and apoptosis of astrocytes and the inhibition mechanism of Rho-associated coiled coil-forming protein kinase(RhoA/ROCK) signaling pathway. Methods Astrocytes were cultured under hypoxia condition and pretreated with exogenous H2S donor, NaHS(50,100,200 μmol/L), ROCK inhibitor Fasudil(5 μmol/L), and co-application of NaHS(200 μmol/L) with Fasudil(5 μmol/L). The cell viability was detected by using cell counting kit-8(CCK-8), the activity of lactate dehydrogenase(LDH) in the culture supernatant was detected by spectrophotometry according to the procedures provided by the assay kit. Apoptosis was detected by flow cytometry, the variation of intracellular free Ca2+fluorescence was detected by fluorescence enzyme labeling. The expression of glial fibrillary acidic protein(GFAP), RhoA, ROCK1and ROCK2protein was detected by Western blot method. Results 100, 200 μmol/L NaHS significantly increased the astrocytes viability cultured under hypoxia condition, and decreased the cell apoptosis, Ca2+fluorescence and LDH activity released from astrocytes into the culture supernatant, suggesting that the protective effect of NaHS on the astrocytes against the hypoxia injury. In addition, Fasudil, an inhibitor of ROCK had the similarly protective effect on the injured astrocytes. In addition, 100, 200 μmol/L NaHS and Fasudil significantly decreased the expressions of GFAP, RhoA, ROCK1and ROCK2in astrocytes, and co-application with Fasudil could significantly enhance the protective effect of NaHS on the injured astrocytes and inhibition of the GFAP, ROCK1and ROCK2expressions. Conclusion H2S has an obviously protective effect on astrocytes against hypoxia injury, which may be related to the inhibition of the activation of RhoA/ROCK pathway.