To investigate the protective effect of acupuncture on the reproductive function of mice by observing the effect of the“Zhibian（BL54）-to-Shuidao（ST28）”acupuncture method on key factors in the death receptor pathway of testicular tissue in asthenospermia mice.

Thirty C57BL/6 male mice were randomly divided into a blank group，a model group and an acupuncture group（n＝10）. The blank group was given an equal volume of 0.9% sodium chloride solution by gavage，while the model group and acupuncture group were given intraperitoneal injection of cyclophosphamide（50 mg·kg^-1·d^-1）for 5 days. After the completion of modeling，the acupuncture group were given continuous acupuncture treatment with“Zhibian（BL54）-to-Shuidao（ST28）”for 2 weeks，once a day，20 minutes for each time. The other two groups were only fixed，and the samples were taken the next day after treatment. In daily observations，the general condition of mice was assessed，and the changes in body weight and testicular and epididymal indices were recorded. The sperm quality of the mice was evaluated. Enzyme-Linked Immunosorbent Assay（ELISA）was used to measure the levels of testosterone（T），follicle-stimulating hormone（FSH）and luteinizing hormone（LH）in the serum. Hematoxylin-Eosin（HE）staining was employed to examine the pathological morphology of testicular tissue. Terminal deoxynucleotidyl transferase dUTP nick end labeling（TUNEL）staining was used to observe cell apoptosis in testicular tissue. Fluorescent quantitative polymerase chain reaction（PCR）and Western blot were employed to assess changes in the mRNA and protein expressions of Fas cell surface death receptor（Fas），Fas ligand（FasL），tumor necrosis factor alpha（TNF-α），TNF receptor 1（TNFR1），tumor necrosis factor-related apoptosis-inducing ligand（TRAIL），death receptor 5（DR5），Caspase-8 and Caspase-3 in testicular tissue. Immunofluorescence staining was conducted to detect the positive expressions of cleaved Caspase-3 and PARPin testicular tissue.

Compared with the control group，the model group exhibited a decrease in body weight of male mice（P<0.01），a decrease in testicular and epididymal mass and coefficients（P<0.01），a decrease in total sperm count，sperm motility and sperm viability（P<0.01），a decrease in serum levels of testosterone（T），follicle-stimulating hormone（FSH）and luteinizing hormone（LH）（P<0.01），deterioration in the pathological morphology of testicular and epididymal tissues，an increase in the number of positively stained apoptotic cells（P<0.01），an increase in the mRNA and protein expression levels of FasL，Fas，TNF-α，TNFR1，FADD，Caspase-8 and Caspase-3（P<0.01，P<0.05）. There was no significant difference in the expression levels of TRAIL，DR5 mRNA or protein（P>0.05），and an increase in the expressions of cleaved Caspase-3 and PARP（P<0.01）. Compared to the model group，the acupuncture group showed no significant change in body weight after treatment（P>0.05）. There was an increase in testicular and epididymal mass and coefficients（P<0.01）. The levels of total sperm count，sperm motility and sperm viability increased（P<0.01）. The serum levels of testosterone（T），follicle-stimulating hormone（FSH）and luteinizing hormone（LH）increased（P<0.01）. There was an improvement in the pathological morphology of testicular and epididymal tissues，a decrease in the number of positively stained apoptotic cells（P<0.01），a decrease in the mRNA and protein expression levels of FasL，Fas，TNF-α，TNFR1，FADD，Caspase-8 and Caspase-3（P<0.01，P<0.05）. There was no significant difference in the expression levels of TRAIL，DR5 mRNA and protein（P>0.05）or decrease in the expression of cleaved Caspase-3 and PARP（P<0.01）.

The“Zhibian（BL54）-to-Shuidao（ST28）”technique can improve spermatogenic cell apoptosis in oligospermia mice and enhance reproductive function，and its mechanism may be associated with the regulation of key factors in the testicular tissue death receptor apoptosis pathway.