Salvia Miltiorrhiza Root (Danshen) Water-Extract Exerts A Protective Effect In Mice With Cyclophosphamide-induced Ovarian Damage

ObjectiveTo investigate the damage and oxidative stress effects of cyclophosphamide?CTX?on mice ovarian function,and whether salvia miltiorrhiza root water-extract?SM?could effectively protect ovarian function and attenuate the oxidative stress of ovarian damage induced by cyclophosphamide.MethodsA total number of thirty female mice?C57BL/6J?were studied at 7 weeks of age,and randomly divided into three groups?n=10 per group?.Mice in each group received premeditated drug treatment.Salvia miltiorrhiza root water-extract?SM?therapeutic group?CTX+SM?:intragastrical?i.g.?administration of Salvia Miltiorrhiza Root water-extract?contain a dose of 0.02g Salvia Miltiorrhiza powder/150?l?daily for 41 days.Mice in control group?CON?and CTX group?CTX?:intragastrical?i.g.?administration of same volume of drinking water.On the 11^th day,6 hours after intragastrical?i.g.?administration,mice in CTX group?CTX?and therapeutic group?CTX+SM?were recived intraperitoneal injection?i.p.?of cyclophosphamide?CTX;80mg/kg·BW;Baxter Oncology GmbH?.For comparison,the same voulm of normal saline?NS?injected into the control group?CON?.During the treatment,body weight of mice measured in every 3 days.Ovarian morphology and follicle number count of HE staining of ovarian tissue sections were observed,vaginal smear and serum anti-Mullerian hormone?AMH?concentration of mice were conducted to learn the damage effects of cyclophosphamide?CTX?on ovarian function,and whether salvia miltiorrhiza root water-extract?SM?could effectively attenuate the damage while restore the ovarian function.Then,according to the superoxide dismutase?SOD?serum level and the malondialdehyde?MDA?serum level of mice,the ovarian oxidative stress induced by cyclophosphamide were studied,and whether salvia miltiorrhiza root water-extract?SM?could attenuate the damage were researched.ResultsCompared with control group?CON?,according to daily vaginal smear after the day of injection?i.p.?of cyclophosphamide?CTX?,mice with injection of cyclophosphamide presented irregular or prolonged estrous cycle.Mice were sacrificed on day 42.Ovrian index was culculated and compared to control group?0.49±0.06mg/g·BW?mice treated with cyclophosphamide showed a much lower ovrian index?0.31±0.05mg/g·BW,P<0.0001?.HE staining of ovarian tissue sections showed that ovarian morphology of mice with the treatment of cyclophosphamide?CTX?was irregular or reduced in total volume,compared with control group?CON?each stage of healthy follicles?including primordial follicles,primary follicles,secondary follicles and antral follicles?were decreased?P<0.05?,and the number of atretic follicles were increased numerously?P<0.01?.The serum concentrations of AMH of mice in cyclophosphamide group?1.54±0.30ng/mL,P<0.05?were also significantly decreased comparing with the control group?2.42±0.15ng/mL?.In addition,it was found that the MDA serum level of mice in CTX group increased obviously?12.69±1.22nmol/mL,P<0.05?,and the SOD serum level of CTX group decreased significantly?847.66±119.66U/Ml,P<0.05?.On the contray,it was showed that ovarian index greatly increased after the administration of SM?0.40±0.06mg/g·BW,P<0.001?.In comparision with mice in CTX group,ovarian morphology of additional SM treated group showed a certain degree of recovery,the relative primordial follicle count increased up to34.75%?P<0.05?,and atretic follicle count decreased significantly?P<0.01?.Serum concentration of AMH in additional SM treated group also increased?2.90±0.24ng/mL,P<0.01?.Furthermore,it was showed that the SOD serum level increased in CTX+SM group?1143.97±77.09U/mL,P<0.05?,and the MDA level presented a downward trend with a concentration of 11.60±1.39nmol/mL?P=0.5908?.ConclusionCyclophosphamide?CTX?induced oxidative stress on C57BL/6J mice,and damaged both ovarian function and preservation.Treatment with SM additionaly could increase mice SOD level,alleviate ovarian injury and restore ovarian function,which indicate that administration of SM could effectively attenuate the oxidative stress induced by cyclophosphamide and protect ovarian preservation in mice.