Objective To investigate the application of three ultrasound-guided nerve blocks in conjunction with general anesthesia in laparoscopic cholecystectomy (LC). Methods Sixty patients taking selective LC at Yulin Hospital, First Hospital, Xi 'an Jiaotong University from October 2022 to November 2023 were selected for the randomized controlled trial, and were divided into an erector spinae plane block (ESPB) group, a paravertebral nerve block (PVB) group, and a transversal-midpleural nerve block (MTPB) group by the random number table method, with 20 cases in each group. All the patients received ultrasound-guided nerve blocks combined with general anesthesia. There were 9 males and 11 females in the ESPB group; they were (49.85±10.58) years old. There were 8 males and 12 females in the PVB group; they were (54.40±10.51) years old. There were 7 males and 13 females in the MTPB group; they were (49.95±11.61) years old. The intraoperative remifentanil and propofol dosages, intraoperative and postoperative mean arterial pressures (MAP), scores of Visual Analogue Scale (VAS), and levels of C-reactive protein (CRP), norepinephrine concentrations, and blood glucose levels at various time points after surgery, as well as postoperative adverse reactions, were recorded and compared. t, F, and χ2 tests were applied. Results There were no statistical differences in the propofol dosage, MAP's before and after the surgery, and score of VAS 24 h after the surgery between the 3 groups (all P>0.05). The remifentanil dosages, MAP's during the surgery, and scores of VAS 2 and 8 h after the surgery in the MTPB group and the PVB group [(1.09±0.50) mg, (89.96±4.56) mmHg (1 mmHg=0.133 ) kPa , (4.95±3.14) , and (3.55±2.50) ; (0.84±0.47) mg, (90.42±5.08) mmHg, (4.60±2.56) , and (4.00±2.55) ] were lower than those in the ESPB group [(1.77±0.29) mg, (93.00±3.90) mmHg, (7.75±1.89) , and (5.95±2.87) ], with statistical differences (F=25.04, 4.06, 8.94, and 4.65; all P<0.05). There were no statistical differences in the blood glucose levels 2, 8, and 24 h after the surgery between the 3 groups (P>0.05). Two, 8, and 24 h after the surgery, the levels of CRP and NE in the MTPB group and the PVB group [(2.83±0.82) ng/dl, (3.34±0.99) ng/dl, (2.75±1.10) ng/dl, (361.89±44.43) ng/L, (351.01±56.17) ng/L, and (358.16±43.28) ng/L; (1.84±0.93) ng/dl, (2.14±1.09) ng/dl, (1.70±0.90) ng/dl, (303.31±55.97) ng/L, (302.45±47.87) ng/L, and (297.46±44.68) ng/L] were lower than those in the ESPB group [(3.87±1.15) ng/dl, (3.89±0.82) ng/dl, (3.92±1.11) ng/dl, (400.77±55.38) ng/L, (407.11±58.59) ng/L, and (403.27±51.48) ng/L], with statistical differences (F=21.62, 16.96, 22.75, 17.67, 18.54, and 25.94; all P<0.05). There was no statistical difference in the incidence of adverse reactions between the 3 groups (P>0.05). Conclusion Application of MTPB or PVB combined with general anesthesia in patients taking LC provides effective postoperative analgesia, reduces intraoperative remifentanil dosage, prolongs analgesic effect, inhibits postoperative stress responses without causing serious adverse reactions, but MTPB is simple and safe.