Experimental Study On Reproductive System Induced By Apigenin In Male Mice

Objective To explore the effect of apigenin on reproductive system in male mice, so as to provide the basic scientific materials for the utilization of apigenin.Methods (1) 84 healthy adult male mice of Kunming strain in SPF degree were divided randomly into seven groups,including negative control group,solvent control group,the 5 experimental groups of apigenin.12 mice of each group were administered by intraperitoneal injection at a certain time every day,negative control group with 0.4ml of normal saline,solvent control group with 0.4 ml of 0.5% DMSO and experimental groups with 0.4ml of apigenin solutions of 5mg/kg,10mg/kg,15mg/kg,20mg/kg and 25mg/kg respectively.After 7 days all mice were weighted and killed,drawn materials and observed their sperm morphology. (2) 100 male mice of Kunming strain in SPF were divided randomly into 5 groups according to body weight (negative control group;solvent control group; 84mg/kg,168mg/kg and 252mg/kg group of apigenin).Each dose group was randomly divided into two groups 7 days and 14 days group.The two groups of mice were given with the corresponding dose by intragastric administration at a certain time every day until 7 and 14 days.After 35 days of first gavage,all mice were weighted,killed,drawn materials and observed their sperm morphology. (3) Observation Indexes:①Liver, spleen, kidney, testis and epididymis were taken to be weighed and calculated organic indexes respectively;②the sperm motility parameters were measured by computer assisted semen analyzer (CASA).③Sperm abnormality test;④Radioimmunoassay of serum hormone (T), follicle-stimulating estrogen (FSH) and luteinizing hormone (LH).⑤Hemoglobin.⑥The content of ALT, AST, ALT/AST, ALB, GLO, A/G, ALP,GGT.⑦Flow cytometry was made to detect all kinds of ploidy cells of testicular spermatogenic cells and its cycle.⑧Pathological alterations of testis in male mice were observed with microscope and ultramicrostructure with electronmicroscope.Results (1)The test of intraperitoneal injection instructions:compared with the negative control group,the ALP,A/G of 5mg/kg group were increased and the GLO,ALT/AST,TG were decreased;the content of haploid cell and the ratio of 1C and 4C testis cells increased significantly in 5 mg/kg group;the sperm density decreased in 25 mg/kg group.(2)The test of intragastric administration showes:at 7 days,compared with the negative control group kidney coefficient of 168mg/kg group and spleen coefficient of 84mg/kg and 252mg/kg groups decreased;the TG of three dose groups of apigenin decreased;the AKP and LDH of the groups decreased,At 14 days,GGT of 168mg/kg increased,168mg/kg and 252mg/kg groups of the ratio of 1C and 2C testis cells increased significantly;VAP, VSL, STR, WOB, b-class proportion of sperm and sperm energy of 252mg/kg group were lower than the negative control group,BCF is higher than the negative control group;with the intragastric prolonging of time,the density and activity of three dose groups mice sperm decearsed;AKP in 84mg/kg and 252mg/kg groups at 14days increased,ACP and LDH in 168mg/kg and 252mg/kg groups decreased. With increasing of dose and treatment time,the ultra-microstructure damage of seminiferous tubules and seitoli cells became more and more serious.the morphology of spermatoblast and the quantity of sperm were changed.Conclusion Apigenin had some effect on the liver function, sperm motility, cell cycle and structure of testis,organizational activity;but teratogenicity in mice sperm of apigenin was not obviouse.