To explore the effects of vaginal delivery and cesarean section on the levator ani muscle morphology, pelvic floor muscle strength, and vaginal mucosal nerve distribution of puerperae. Methods: 354 puerperae were selected as the research subjects between December 2018 and December 2020. According to the delivery methods, these puerperae were divided into group A (248 puerperae with vaginal delivery) and group B (106 puerperae with cesarean section). In the 42th day after delivery, the levator ani muscle morphology, the pelvic floor muscle strength, and the vaginal mucosal nerve distribution of the puerperae were compared between the two groups. Results: In the rest state, there were no significant differences in the horizontal contained angle, diameter, area, and transverse diameter of the reproductive hiatus of the puerperae between the two groups (P>0.05). In the contraction state, the value of the horizontal contained angle reproductive hiatus of the puerperae in group A was significantly lower than that of the puerperae in group B, and the values of the diameter, the area, and the transverse diameter of reproductive hiatus of the puerperae in group A were significantly higher than those of the puerperae in group B (P<0.05). In Valsalva state, there was no significant difference in the horizontal contained angle of the reproductive hiatus of the puerperae between the two groups (P>0.05). The values of the diameter, the area, and the transverse diameter of the reproductive hiatus of the puerperae in group A were significantly higher than those of the puerperae in group B (P<0.05). The changes of the values of the horizontal contained angle, the diameter, and the area, and the bladder neck movement of the puerperae in group A were significantly less than those of the puerperae in group B in the rest and contraction states (P<0.05). The muscle strength of type Ⅰ or type Ⅱ muscle fibers of the puerperae in group A was significantly lower than that of the puerperae in group B (P<0.05), and there was no significant difference in the fatigue of type Ⅰ and type Ⅱ muscle fibers of the puerperae between the two groups (P>0.05). The distribution of nerve fiber distribution status of vasoactive intestinal peptide (VIP) and neuropeptide Y (NPY) in the anterior vaginal wall lamina propria of the puerperae in group A was significantly less than that of the puerperae in group B, and the distribution of 9.5 protein gene products and VIP nerve fibers in the posterior cortical lamina propria of the puerperae in group A was significantly less than that of the puerperae in group B (all P<0.05). There were no significant differences in the distributions of the other vaginal nerves of the puerperae between the two groups (P>0.05). Conclusion: Both vaginal delivery and cesarean section will damage the pelvic floor function of the puerperae, thus it is necessary to strengthen the protection of the pelvic floor of the puerperae.