Studied On Testosterone Synthesis Effects Of Carbamazepine In Rats Based On ERK1/2-CREB Signaling Pathway

Objective To observe the effect of carbamazepine on ERK1/2-CREB signaling pathway in normal male rats,and to explore the mechanism of its effect on testosterone synthesis in rats.Methods120 male SD rats aged from 6 to 8 weeks with body weight of 200～220g were randomly divided into normal control group and carbamazepine 50,100,200 and 400 mg/kg groups with 24 rats in each group.Each group was given intragastric administration according to dose for consecutive 90 days,while the normal group was given distilled water of equal volume.After the last administration,the animals were fasted without water for 12 hours,and 3% chloral hydrate was used to anesthetize the animals.Blood was collected from the abdominal aorta,and organs such as testicles and epididymis were extracted.Calculate the organ coefficient,detect the sperm viability and vitality;Enzyme linked immunosorbent assay(ELISA)is used to detect blood and Testoserone(T),Luteinizing hormone(LH),follicle-stimulating hormone(FSH),Estradiol(E2),Prolactin(Prolactinin testicular tissue.PRL)and Progesterone(P).Hematoxylin-eosin staining(HE)was used to observe the pathological staining of testicular tissue.The expression of STAR,3β-HSD and 17β-HSD in testis was observed by Immunohistochemistry staining.The expressions of STAR,3β-HSD,17β-HSD,ERK1/2,CREB,p-ERK1/2 and p-CREB in testis were detected by Western blot(WB),and the mRNA expressions of ERK1/2 and CREB in testis were detected by Real time quantitative polymerase chain reaction(RT-qPCR).Results1.Effect of carbamazepine on body weight of rats: compared with the normal group,testicular weight and organ coefficient of testis were decreased in each group,and the decrease was significant in the 200 and 400mg/kg groups,with statistical significance(P < 0.05).2.Effects of carbamazepine on sperm quality and testis pathology in rats: compared with the normal group,sperm motility rate and sperm motility in carbamazepine groups were significantly decreased,with statistical significance(P < 0.05),and the sperm motility rate and sperm motility in the 400mg/kg group were the most significantly decreased.3.Effects of carbamazepine on serum and testis sex hormones in rats: The results of six serum sex hormones levels showed that compared with the normal group,T levels in each group were significantly decreased,and showed a step-down trend with the increase of drug concentration,with statistical significance(P < 0.05);The levels of LH and FSH were significantly increased,and the differences were statistically significant(P < 0.05).The results of six levels of sex hormones in testis tissue showed that compared with the normal group,the levels of T and P in the administration groups were significantly decreased,and showed a gradual decline with the increase of drug concentration,with statistical significance(P < 0.05).The levels of LH and FSH were significantly increased in each group,and the differences were statistically significant(P < 0.05).4.Effects of carbazepine on the expression of STAR,3β-HSD,17β-HSDERK1/2,CREB: Compared with the normal group,the protein expressions of STAR,3β-HSD,17β-HSD,ERK1/2 and CREB in the testis of rats were significantly decreased in the treatment groups.5.Effects of carbamazepine on the expressions of ERK1/2 and CREBmRNA in rats:Compared with the normal group,the expressions of ERK1/2 and CREBmRNA in the treated groups were significantly decreased.ConclusionCarbamazepine for normal male rat reproductive system has obvious toxicity,one of its mechanism may be through regulating ERK1/2-after CREB signaling transduction,suppresses the mesenchymal cells testosterone synthesis of related enzyme protein expression decrease testosterone synthesis,thus stress HPT shaft production has changed the hormone levels and the relevant receptors,eventually led to the decrease of the rat sperm quality.