| Objective:To investigate the effect of Coridius chinensis extract（CcE）on the testicular injury in rats due to manganese exposure and its mechanism by detecting the expression of FAK,Src as well as TJ proteins in the PI3K/c-Src/FAK signaling pathway.Methods:The sixty adult Sprague Dawley（SD）male rats were randomly divided into the control group,the model group,C.chinensis intervention groups（low-,medium-,high-）and the positive group.The model group were injected intraperitoneally with MnCl2solution（15mg/kg,30d）,C.chinensis intervention groups（low-,medium-,high-）were given CcE（50mg/kg,100mg/kg,200mg/kg）and the positive group was given Jin kui shen qi wan solution（1.25g/kg）by intragastric administration once a day for 30 days with MnCl2 solution 15mg/kg simultaneously.1.The reproductive ability index detection measures the sperm number,organ coefficient of testis and accessory sex glands（epididymis,seminal vesicle and prostate,levator ani muscle,preputial glands）;2.Behavioral observation was used to observe the mating behavior index of male rats in15min,including latency and frequency of capturing,mount,intromission and ejaculation;3.The morphological structure of the testicular tissue were observed by light and transmission electron microscope;4.The changes in the expression of Fak,c-Src and Claudin1 m RNA in the testicular tissue was detected by qRT-PCR;5.The expression of p-FAK,p-Src and TJ proteins（Claudin1,Occludin,ZO-1 and JAM-A）in the testicular tissue was detected by Western Blot.Results:1.The effect of C.chinensis on the sperm count,the coefficient of the testis and accessory sex glands in the rats exposed to manganese:Compared with the control group,the sperm count,the organ coefficient of testis and epididymis as well as preputial glands were decreased in the model group significantly（P<0.05）;The sperm count,the organ coefficient of the testis and accessory sex glands were increased in C.chinensis intervention groups（low-,medium-,high-）,in addition,the sperm count and the organ coefficient of the levator ani muscle and preputial glands were increased significantly in C.chinensis medium-intervention group（P<0.05）.2.The effect of C.chinensis on the mating behaviors of male rats of exposed manganese:the latency of capturing,mount,intromission and ejaculation were prolonged in the model group significantly（P<0.05）,moreover,the frequency were decreased significantly（P<0.05）;the results showed that the latency of mating behaviors were shortened obviously and the frequency were increased significantly in C.chinensis intervention groups（low-,medium-,high-）（P<0.05）.3.The effect of C.chinensis on the morphological structure of the testicular tissue of male rats of exposed manganese:HE results show broadened interstitial testicular tissue,larger vacuoles between seminiferous tubules,and the irregular arrangement of spermatogenic cells,decreased number and reduced layers of spermatogenic cells in lumen were also found in the model group;meanwhile,in C.chinensis intervention groups,the testicular morphological structures were obviously improved contained the broadened interstitial testicular tissue and the number as well as the layers of sperm spermatogenic cells in lumen.TEM results show that in the model group,spermatogonium atrophy and the lacuna widened between the Sertoli cells and the Spermatogonium,the mitochondrial showed an obvious abnormity,swelling,vacuoles and loss of crests,the endoplasmic reticulum was significant expansion.additionally,the destroyed integrity of BTB,there were no visible BTB-associated junctions.Meanwhile,compared with the model group,the swelled mitochondrial and the expanded endoplasmic reticulum as well as the impaired BTB of the Sertoli cells were improved significantly in C.chinensis intervention groups.4.The effect of C.chinensis on the PI3K/c-Src/FAK signal pathway of male rats of exposed manganese:The relative expression levels of Fak and c-Src mRNA were up-regulated and that Claudin1 mRNA was down-regulated significantly in the model group compared with the control group（P<0.05）.Meanwhile,the relative expression of p-FAK protein was raised while the relative expression of TJs were reduced with significant difference（P<0.05）.Since C.chinensis intervention inhibited the relative expression of Fak mRNA,the relative expression of p-FAK was decreased significantly compared to the model group（P<0.05）,while the relative expression of Claudin1 mRNA and TJs were increased significantly（P<0.05）,the results showed that C.chinensis can improve the micro-environment of spermatogenesis.Conclusions:1.C.chinensis can improve the reproductive ability and mating behavior of the manganese-exposed rats.2.C.chinensis can improve significantly the morphological structure of Sertoli cells and so as to enhance the protection.3.The effect of C.chinensis on the manganese-exposed rats and its mechanism might be inhibiting the expression of activated FAK of the manganese-exposed rats through dephosphorylatiing of p-FAK and increasing the expression of TJs in the PI3K/c-Src/FAK signal pathway. |
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