Objective To explore the regulatory effect of miR-181a on PTEN-induced kinase 1 (PINK1)/Parkin-related genes (Parkin) pathway, and on mitochondrial autophagy of osteoclasts in osteoporosis (OP) rats. Methods Twenty healthy female SD rats were randomly divided into osteoporosis (OP) model group and control group (10 each). Rats in OP model group were employed to prepare OP model. Osteoclasts were extracted from OP rats, and set them as: OP group (not transfected), si-miR-181a group (transfected with si-miR-181a vector plasmid), si-NC group (transfected with negative control plasmid), ad-miR-181a group (transfected with ad-miR-181a vector plasmid), and ad-NC group (transfected with negative ad-NC control plasmid),and the osteoclasts of rats in control group were cultured normally and set as normal control group. The expression of miR-181a was detected by RT-PCR, and MTT and flow cytometry were performed to detect the survival and apoptosis of osteoclasts, the mitochondria autophagy was observed with transmission electron microscope, the expression of Parkin in mitochondria was detected by immunofluorescence co-localization, Western blotting was performed to detect the expression of PINK1/Parkin and autophagy,as well as apoptosis related proteins. Knockdown of miR-181a in osteoclasts of OP rats to down regulate the expression of Parkin and verify the reversal effect of Parkin on miR-181a. Double Luciferase Report experiment verified the targeted regulation between miR-181a and Parkin. Results Compared with normal control group, miR-181a (1.59±0.15 vs. 1.02±0.11), Parkin⁺TOMM2⁺(2.02±0.20 vs. 0.13±0.10), Parkin (1.83±0.18 vs. 1.13±0.10) in osteoclasts of OP rats, and PINK1 (1.93±0.19 vs. 1.03±0.10)expression and survival rate (157.06%±12.32% vs. 100.09%±0.05%), autophagy marker protein-LC3-Ⅱ/Ⅰ ratio (1.89±0.18 vs. 1.15±0.10) increased (P<0.05). Compared with OP group, after up-regulating the expression of miR-181a in osteoclasts of OP rats, the cell survival rate (222.96%±22.15%) increased, Parkin⁺TOMM2⁺ (1.01±0.11), LC3-Ⅱ/Ⅰ ratio (1.36±0.12) and apoptosis rate (3.28%±0.35%) decreased (P<0.05). While after down-regulating the expression of miR-181a in osteoclasts of OP rats, the cell survival rate (106.96%±10.15%) decreased, Parkin⁺TOMM2⁺ (2.97±0.29), LC3-Ⅱ/Ⅰ ratio (2.47±0.24) and apoptosis rate (19.71%±1.83%) increased (P<0.05). The dual luciferase reporter test showed that Parkin is the target gene of miR-181a. Down-regulating Parkin expression can reverse the effects of miR-181a low expression in promoting mitochondrial autophagy and inhibiting cell survival (P<0.05). Conclusion Up-regulation of miR-181a expression in OP rat's osteoclasts can target down-regulation of Parkin expression, inhibit activation of mitochondrial autophagy, and promote the survival of osteoclasts.