Effects Of Dendrobium Officinale Polysaccharide On Cyclophosphamide-induced Reproductive Damage In Male Mice

Background:Cyclophosphamide?CP?is a non-special cytotoxicity drugs which is widely used in the treatment of various tumors,including genital tumors.However,its side effects can damage reproductive function.How to effectively alleviate the toxicity of chemotherapy to reproductive function is one of the important contents of cancer treatment.Dendrobium officinale Kimura et Migo is a precious traditional Chinese medicine in China.Polysaccharide?DOP?from Dendrobium officinale Kimura et Migo is the main bioactive component,it has multiple beneficial effects such as anti-tumor,anti-oxidation,anti-aging and immunoregulation.However,the medicinal effects of DOP on reproductive function have not been reported.Objective:The aims of this study were research the change of DOP?200mg/kg?on CP-induced reproductive damage in male mice and analyze the effects of DOP on CP-induced damage to reproductive system in male mice.On this basis,we explore the mechanism of DOP in preventing and treating on CP-induced reproductive damage.Methods:?1?6-week-old male mice were randomly divided into 3 groups?n=10?,CK group?distilled water+saline+distilled water?,CP group?distilled water+CP+distilled water?and DOP group?DOP+saline+DOP?.Mice of CK and CP groups were orally given distilled water for7 days,then saline or CP were injected intraperitoneally on the eighth day with single dose,followed with orally gavage of distilled water for another 7 days.After 14 days of gavage period,mice were killed.In the DOP group,mice were orally given DOP for 7 days,and subsequently saline injected on eighth day,followed with orally gavage of DOP for another 7 days with 14 days of gavage period.After last gavage,the mice were killed.Body weight,testis weight and epididymis weight were weighed to calculate the coefficients of testis and epididymis.Pathological changes in testis and epididymis were observed with microscope.Quality of sperm was analyzed.Level of MDA,SOD,CAT,GSH-Px and GSH of testis were detected.?2?6-week-old male mice were randomly divided into 9 groups?n=10?.Of 7-days groups,CK_7d group?distilled water+saline?,CP_7d group?distilled water+CP?,and CP+DOP_7d group?DOP+CP?were grouped.Of 14-days groups,CK_14d group?distilled water+saline+distilled water?,CP_14d group?distilled water+CP+distilled water?and CP+DOP_14d4d group?DOP+CP+DOP?were grouped.Of 21-days groups,CK_21d group?distilled water+saline+distilled water?,CP_21d group?distilled water+CP+distilled water?,and CP+DOP_21d1d group?DOP+CP+DOP?were grouped.Mice of CK groups and CP groups were orally given distilled water for 7 days,single dose of saline or CP was injected intraperitoneally on the eighth day respectively.The mice were killed on the next day or after another 7 days/14 days of distilled water.According to the days of gavage,the groups were labeled as CK_7d,CK_14d,CK_21d and CP_7d,CP_14d,CP_21d respectively.Mice of CP+DOP groups were orally given DOP for 7 days,single dose of CP was injected intraperitoneally on the eighth day.The mice were killed on the next day or after another 7 days/14 days of DOP.The groups were labeled as CP+DOP_7d,CP+DOP_14d and CP+DOP_21d1d respectively.Body weight,testis weight and epididymis weight were weighed to calculate the coefficients of testis and epididymis.Pathological changes in testis and epididymis were observed with microscope.Quality of sperm was analyzed.Level of MDA,SOD,CAT,GSH-Px and GSH of testis were detected.Results:?1?In the DOP group,the mice grew normally as control group,the body weight,testis and epididymis coefficients did not showed significantly difference in comparison to mice of control group.The numbers of spermatogenic cells,Sertoli cells and Leydig cells in testis were normal.There were a large numbers of sperms in the epididymis,and there was no obvious pathological damage in testis and epididymis.There was no significant decrease in sperm quality,and no oxidative damage in testis.There was no significant difference in reproductive system between the DOP group and CK group?P>0.05?.After CP treatment,the weight of mice was decreased dramatically,the coefficients of testis and epididymis were decreased.The numbers of spermatogenic cells,Sertoli cells and Leydig cells in testis were decreased,and the numbers of sperms in epididymis were decreased.The analysis of sperm in epididymis tail showed that total sperm numbers,sperm motility and rate of acrosome reaction were significant decreased.The rate of sperm malformation was increased greatly.The obvious oxidative damage was observed in testis.?2?Compared with corresponding CP_7d/CP_14d/CP_21d group,the coefficients of testis and epididymis were increased in CP+DOP_7d group,CP+DOP_14d group and CP+DOP_21d group.Pathological observation showed that the numbers of spermatogenic cells,Sertoli cells and Leydig cells in testis were increased,and the numbers of sperms in epididymal were increased.There were significant differences in sperm numbers,sperm motility,the rate of acrosome reaction and sperm malformation in three CP+DOP groups(CP+DOP_7d group,CP+DOP_14d group and CP+DOP_21d)compared with corresponding CP groups?P<0.05?.After CP injection,MDA content was increased,activity of SOD,CAT,GSH-Px and GSH content was decreased.The MDA content was the highest,activity of SOD,CAT,GSH-Px and GSH content were the lowest with CP_21d group.CP and DOP co-treatment could decrease MDA content and increase activity of SOD,CAT,GSH-Px and GSH content.The MDA content,SOD,CAT,GSH-Px activity and GSH content in CP+DOP_21d group were significantly different from those in CP_21d1d group?P<0.05?.Conclusions:?1?DOP did not affect the growth and development of reproductive system in male mice,including the development of testis and epididymis.DOP treatment showed no adverse effects on the quality of sperm.DOP treatment showed no toxicological effects on reproductive system in male mice as well.?2?DOP can effectively alleviate CP-induced reproductive toxicity in testis and epididymis in male mice.DOP oral treatment can prevent and repair the sperm toxicity induced by CP.The mechanism is associated with regulatory effects of DOP on balance status of oxidation-antioxidation in male reproductive system.