Effects of Manual Acupuncture on Malondialdehyde Levels and State Anxiety Inventory Scores in Active Smokers: A Randomized Controlled Clinical Trial

Cigarette smoking is an important source of reactive oxygen species and may contribute to oxidative stress through increased lipid peroxidation and an imbalance between oxidant production and antioxidant defense. Malondialdehyde (MDA), a product of lipid peroxidation, is commonly used as a biomarker of oxidative stress. In addition to its biological effects, smoking and temporary smoking abstinence may also be associated with anxiety and other psychological symptoms.

Acupuncture has been proposed as a non-pharmacological intervention that may modulate oxidative stress and anxiety through neurophysiological, autonomic, neuroendocrine, and antioxidant-related mechanisms. However, evidence regarding the effects of a single session of manual acupuncture on MDA and state anxiety in active smokers, particularly within a 90-minute post-intervention period, remains limited. This study also aims to examine the correlation between changes in MDA levels and changes in SAI scores, in order to explore the relationship between oxidative stress and state anxiety following the intervention.

This study is designed to evaluate the effects of manual acupuncture on MDA concentration and SAI scores in active smokers. The intervention will use bilateral HT7 Shenmen, ST36 Zusanli, and SP6 Sanyinjiao, together with GV20 Baihui.

This study is a randomized controlled trial with two parallel groups. A total of 44 participants will be recruited and randomly allocated in a 1:1 ratio to either a manual acupuncture group or a sham acupuncture group, with 22 participants in each group. Participants will be blinded to treatment allocation, and outcome assessment and data collection will be performed by an independent investigator who is not involved in administering the intervention.

Participants will be active smokers aged 21-40 years who smoke at least 20 cigarettes per day and have smoked for at least five years. Participants must have no previous history of manual acupuncture treatment, have a body mass index (BMI) of 18.5-29.9 kg/m², be willing to complete all study procedures and provide written informed consent, be able to communicate effectively, and be willing to complete the study questionnaires independently.

At the study visit, participants will undergo an initial assessment during the two-hour smoking-abstinence period, including medical history, physical examination, questionnaire completion, and placement of an intravenous catheter. After completing the two-hour abstinence period, a baseline blood sample will be collected for measurement of plasma MDA, and the baseline SAI will be completed.

In the intervention group, a Park Sham Needle Device was used with sterile 0.25 × 25 mm acupuncture needles. The needles were inserted perpendicularly to a depth of 20 mm at bilateral ST36, SP6, and HT7, and to a depth of 10 mm at GV20. The needles were retained for 30 minutes. Manual stimulation was performed using a twirling-rotating technique, consisting of alternating clockwise and counterclockwise needle rotation with a rotational amplitude of 180 degrees at a frequency of once per second for 10 seconds, at minute 0 and minute 15. After needle removal, gentle pressure was applied to the insertion sites for 10 seconds to ensure that no bleeding occurred.

In the control group, the same Park Sham Needle Device was used; however, the needle was blunt and did not penetrate the skin surface. The needle was placed at the same acupoints and retained for 30 minutes without manipulation.

Both interventions will be performed with participants in a standardized supine position in a controlled treatment room. Skin antisepsis will be performed using 70% alcohol before the intervention. The procedure will be administered by a physician trainee in the Medical Acupuncture Specialist Program with a minimum competency level equivalent to second-year residency or above.

Participants will be reassessed 90 minutes after completion of the intervention. A second blood sample will be collected to determine post-intervention plasma MDA concentration, and the SAI will be repeated to evaluate changes in state anxiety. Blood samples will be collected in EDTA-containing tubes and processed and stored according to the laboratory protocol.

The primary outcome of interest is the change in plasma MDA concentration from baseline to 90 minutes after the intervention and the difference in this change between the manual acupuncture and sham groups. The secondary outcome is the change in SAI score from baseline to 90 minutes after the intervention and the difference in this change between the two groups. An additional exploratory outcome is the correlation between changes in plasma MDA concentration and changes in SAI scores from baseline to 90 minutes after the intervention.

Safety will be monitored throughout the study. Potential acupuncture-related adverse events include local pain, bleeding, bruising or hematoma, dizziness, vasovagal symptoms, and local infection. Potential intravenous catheter-related adverse events, including pain, bleeding, hematoma, and local irritation, will also be monitored. Participants will remain under observation during the study visit, and any adverse events will be documented and managed according to the study protocol.