Effects of Allicin Pretreatment on Anti-oxidation Ability and Delayed Onset Muscle Soreness

Subjective pain sensation was assessed using the Visual Analog Scale (VAS), which ranges from 0 mm (no pain) to 100 mm (worst imaginable pain). Higher scores indicate a worse outcome (greater pain intensity). Measurements were taken to evaluate the degree of Delayed Onset Muscle Soreness (DOMS) at the following time points: pre-medication (Pre-drug), pre-exercise (Pre-exercise), immediately post-exercise (Post-exercise), 24 hours post-exercise (24 h), 48 hours post-exercise (48 h), and 72 hours post-exercise (72 h).

Bilateral calf circumference was measured using a soft measuring tape (Hoechstmass, Germany) at the following time points: before medication (Pre-drug), before exercise (Pre-exercise), immediately after exercise (Post-exercise), 24 hours post-exercise, 48 hours post-exercise, and 72 hours post-exercise. Subjects lay prone on the examination table with legs slightly abducted and knees extended. The tape was placed horizontally around the thickest part of each calf, and the circumference was recorded in centimeters (cm). Each side was measured three times, and the average value was used for analysis.

A vertical jump tester was used to measure the vertical jump height of the subjects at Pre-drug, Pre-exercise, Post-exercise, 24 h, 48 h, and 72 h. Subjects stood naturally on the jump platform with feet slightly apart, ready for testing. When "0.0" appeared on the screen of the electronic vertical jump platform (Hongkangda Sports Technology Development Co., Ltd., Beijing), and the subject heard "start," the test officially began. The subject bent their knees into a half-squat, swung their arms back, and forcefully jumped vertically upward. When the subject landed back on the platform, the display showed the corresponding value. The test was repeated three times, with the highest value taken.

The MicroFET2 handheld muscle strength tester (Hoggan, USA) was used to measure the posterior calf muscle strength of subjects at Pre-drug, Pre-exercise, Post-exercise, 24 h, 48 h, and 72 h. The subjects were in a prone position with arms placed at the sides, and the legs bent at 60° during the initial measurement. The tester placed the device under the Achilles tendon, and the subject resisted by flexing the calf for 8 seconds, then returned to the initial position. Each leg's muscle strength was tested five times, with the highest value recorded.

A goniometer (Changzhou Youbang Medical Rehabilitation Equipment Co., Ltd.) was used to measure the bilateral knee joint range of motion (ROM) in flexion at Pre-drug, Pre-exercise, Post-exercise, 24 h, 48 h, and 72 h. Subjects were prone (with hips and knees extended) on the edge of the examination bed, exposing the knee joint. The tester placed the axis of the goniometer at the fibular head of the knee joint, aligning the stationary arm parallel to the femoral shaft and the moving arm parallel to the fibular shaft. The moving arm followed the motion of the fibula, and the value of the knee joint's ROM was read at the terminal position

At Pre-drug, Pre-exercise, post-exercise, 24 h, 48 h, and 72 h, 4 ml of median cubital vein blood was collected from each subject into an anticoagulant tube. The sample was centrifuged using a benchtop high-speed centrifuge (temperature: 4°C, 1000 rpm/min) for 15 minutes, and the supernatant was transferred to an EP tube and stored at -80°C for further testing. The thiobarbituric acid (TBA) method was used to measure the MDA content, while the T-AOC was determined using the ferric ion reducing/antioxidant power (FRAP) assay. The activity of CK-MM was measured by Enzyme-Linked Immunosorbent Assay (ELISA), and CK (Creatine Kinase) activity was measured using an optimized DGK colorimetric method. The MDA, T-AOC, and CK assay kits were purchased from Nanjing Jiancheng Bioengineering Institute, and the CK-MM assay kit was obtained from CUSABIO® (Wuhan Huamei Bioengineering Co., Ltd.).