| BACKGROUNDDefective sperm function is considered as the major cause of human infertility.As one of the most common causes of male infertility,asthenozoospermia is characterized by impaired sperm motility.Clinically,excessive production of reactive oxygen species by oxidative stress in seminal plasma,and exacerbated ROS-regulated damage of sperm membranes are often observed in asthenozoospermic patients.At present,oral antioxidants are mainly used in the treatment of oxidative stress in western medicine.Given that the etiology and pathogenesis of asthenozoospermia remain obscure,there is still no effective and specific treatment in clinical and researches are needed to delve further into its clinical intervention.The therapeutic effect of moxibustion is a kind of comprehensive effect.Modern researches have shown the moxa combustion products can exhibit multiple effects,such as bactericidal and antiviral,immune regulation,free radical scavenging and antioxidant,lipid metabolism regulation,and inflammatory response reduction.As one product of moxa combustion featuring special aromatic odor,moxa smoke(MS)shows a variety of biological activities and plays a key role to ensure the efficacy of moxibustion treatment.OBJECTIVEIn the current study,we investigated and compared the effect of differentiated concentrations of moxa smoke on sperm parameters and oxidative stress reactions of the reproductive system in the ornidazole(ORN)-induced asthenozoospermic rats to demonstrate its therapeutic effect on reproductive functions.Moreover,we highlighted the alleviating effect of moxa smoke on asthenozoospermia in comparison with cigarette smoke(CS)to provide experimental proofs for the effectiveness and safety of moxa smoke.METHODS72 male SD rats were randomly and evenly divided into six groups(n=12),including vehicle,ORN,ORN+CS,ORN+MS1(low concentration),ORN+MS2(medium concentration),and ORN+MS3(high concentration).Asthenozoospermia model was established by intragastric administration of ornidazole once daily till the end of intervention.Smoke intervention was initiated 11 days after intragastric administration of ORN.For the ORN+MS1,ORN+MS2 and ORN+MS3 groups,the concentrations of moxa smoke remained stable at 0.4%,2%and 15%in each respective moxa group.For the ORN+CS group,the concentration of cigarette smoke was set up at 2%,which was consistent with the ORN+MS2 group.Rats in the smoke intervention groups underwent 20-minute smoke exposure per day,6 days per week for 8 weeks.Eight rats in each group were randomly selected for detection of body weight gain,testis weight,testis index,sperm parameters and oxidative stress levels,whereas the remaining four rats were employed for examining of testicular histopathology.RESULTS1.Body weight gain:the vehicle rats gained more weight,whereas the body weight gain in the other groups decreased in varying degrees.Compared with the vehicle group,no significant difference was found among each group(P>0.05)except for the ORN+MS2 group(P<0.05).2.Testis weight and testis index:the testis weight of rats in vehicle group was the heaviest,while the testis weight in the other groups reduced in varying degrees.Testis weight and testis index showed no obvious difference among the six groups(P>0.05).3.Sperm parameters:the results revealed that the sperm motility and sperm viability had significantly decreased(P<0.01,P<0.01)and the sperm velocity parameters and swimming patterns were also notably diminished(P<0.01 or P<0.05)in the ORN-treated rats,as compared with the vehicle control rats.The deteriorated parameters of sperm motility and sperm viability induced by ORN were restored by low,medium and high concentrations of moxa smoke(P<0.01,P<0.01,P<0.01)and the sperm velocity parameters and swimming patterns were also obviously improved(P<0.01 or P<0.05),in comparison with the ORN-treated group.Notably,the effect was more obvious with the increase of the concentration of moxa smoke in the range of concentration set in this experiment.Except for sperm motility and sperm viability(P<0.05,P<0.05),there was no significant difference in other sperm parameters between the ORN-treated and ORN+CS rats(P>0.05).4.GSH-Px,SOD,CAT,MDA and NOS levels in testis:significant differences were determined in regard to testicular GSH-Px,CAT,MDA and NOS levels between the vehicle group and ORN-treated group.Compared with the vehicle group,the activities of GSH-Px and CAT were notably reduced(P<0.05,P<0.01)while the MDA and NOS levels were remarkably increased(P<0.01,P<0.01)in the ORN-treated group.However,the SOD level in the ORN-treated group was not significantly down-regulated(P>0.05).After the intervention of moxa smoke,the activities of antioxidant enzymes increased to varying degrees,whereas the contents of oxidation products decreased.The main performance showed that low concentration of moxa smoke exerted better effects on increasing SOD and CAT activities(P<0.05,P<0.01)and reducing MDA and NOS contents(P<0.01,P<0.05),medium concentration of moxa smoke did well in improving GSH-Px,SOD and CAT activities(P<0.05,P<0.05,P<0.05)and decreasing NOS content(P<0.05),and high concentration of moxa smoke was good at enhancing the activities of GSH-Px and CAT(P<0.01,P<0.01).Different concentrations of moxa smoke did not manifest a definite concentration-effect rule in improving oxidative stress injury.In comparison with the ORN-treated group,no significant variation in terms of GSH-Px,SOD,CAT,MDA and NOS levels could be determined in the ORN+CS group(P>0.05)5.Histopathological observation of testis:compared with the vehicle group,spermatogenesis of the ORN-treated group was notably reduced.It was manifested by slight atrophy of seminiferous tubules,mildly disordered and loosely connected spermatogenic cells with reduced layers and numbers,slightly widened lumen space,and small decreased size of spermatid cells.The incidence of spermatogenesis hypofunction decreased with the interventions of cigarette smoke and high concentration of moxa smoke,yet improved after the treatment of low and medium concentration of moxa smoke.CONCLUSION1.The deteriorated parameters of sperm motility induced by ORN were restored by differentiated concentrations of moxa smoke.Notably,the effect became more obvious with the increase of the concentration of moxa smoke in the range of concentration set in this experiment.2.Differentiated concentrations of moxa smoke improved the sperm motility and the histopathological manifestations in testis of asthenozoospermic rats by restoring the inhibited activities of antioxidant enzymes and the enhanced levels of oxidation products in vivo.3.Cigarette smoke did not improve the sperm quality and the oxidative stress responses of testis tissue in asthenozoospermic rats,and aggravated the damages of histopathological manifestations in testis,which was different from the effect of moxa smoke. |
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