Objective: The present study investigates the expressions of miR-133b and RhoA in acute ischemic stroke (AIS) patients. Methods: In the present study, serum samples from AIS patients and healthy controls were collected to determine the regulatory miRNAs and RhoA involved in AIS.The expression levels of miR-133b in serum were detected by real-time quantitative PCR, and the expression levels of RhoA in blood were detected by ELISA. The correlation between the levels of miR-133b and RhoA was analyzed. Results: The results indicate that the RhoA level in the AIS patients is very high compared with that in controls. The level of miR-133b showed the opposite change with that of RhoA.The association between the National Institutes of Health stroke scale (NIHSS) scores with RhoA and miR-133b-related RhoA serum levels was calculated using Pearson?s/Spearman?s correlation coefficient. The findings revealed a positive correlation between NIHSS scores and RhoA level, whereas a negative correlation was observed between NIHSS scores and miR-133b. In addition, the relationship between serum RhoA and miR-133b was negative in AIS patients. Conclusion: Serum miR-133b and the miR-133b-regulatory RhoA may serve as molecular markers for AIS.