The Improvement Effect Of Morinda Officinalis Extract On Cytoxan-impaired Testis Of Adult Male SD Rats

Objective:To discuss the improvement effect that Morinda Officinalis extract exert on the spermatogenesis of testis of cytoxan impaired adult male SD rats.Methods:56 adult SD male rats were randomly divided into 7 groups marked as blank control group, CTX model group, CTX+NS group, 10g/kg Morinda Officinalis extract group,20g/kg Morinda Officinalis extract group,30g/kg Morinda Officinalis extract group,40g/kg Morinda Officinalis extract group. After gavage has been given for two weeks, the weight, the testis ratio and epididymis ratio were recorded. The changes of testicular tissue were observed with HE stained optical microscope. Besides, the receptor of FSHR in testicular tissue was labeled with IHC, and the results were observed with optical microscope. Meanwhile, the expression levels of FSHR mRNA and ABP mRNA were determined by real-time fluorescent PCR method. Finally, the expression levels of FSHR receptor were determined by Western-blot method.Results:1. Rat stomachs were perfused with Morinda Officinalis extract, the weight of the CTX model group was not significantly different from that of the blank control group; Compared with the blank control group and the CTX model group, the weight of CTX+NS group, Morinda Officinalis extract 10g/kg group,20 g/kg group,30g/kg group and 40g/kg group were distinctly decreased(P<0.05); The weight of the CTX+NS model group was not significantly different from that of the 10g/kg Morinda Officinalis extract group,20g/kg group,30g/kg group and 40g/kg group.2. The testis ratio:the testis ratio of the CTX control group was lower than that of the blank control group(P<0.05); the testis ratio of 20 g/kg Morinda Officinalis extract group,30g/kg group and 40g/kg group were dramatically higher than that of the CTX+NS group (P<0.05). The epididymidis ratio:the epididymidis ratio of the four treatment groups were not prominently different from CTX+NS group(P>0.05).3. Morphological observation of testis:in CTX model group and CTX+NS group, the epithelium of seminiferous tubule was thinned, spermatogenic cells in the lumen were in disordered level, spermatogenic cells of each level decreased significantly with sparse distribution, spermatoblast and sperms were not easy to be seen in the lumen, and falling necrotic cells could be seen; in the Morinda Officinalis extract lOg/kg group, it could be seen that the structures of most of the seminiferous tubule were significantly ameliorated, the spermatogenic cells in the lumen were hierarchical, the number of spermatogenic cells in each level increased with a close arrangement. More sperms could be seen in most of the lumens, but there were still spermatogenic cells in some seminiferous tubules disordered, with spermatogenic cells decreasing and spermatoblast and sperms not easy to be seen; in the Morinda Officinalis extract 20g/kg group,30g/kg group and 40g/kg group, it could be seen that the structure of seminiferous tubule was basically normal with thickened seminiferous epithelium, hierarchical spermatogenic cells in the lumen, increasing number of spermatogenic cells in each level and close arrangement, and more sperms could be seen in the lumen.4. The results of immunohistochemistry showed that follicle-stimulating hormone receptor was expressed in the testicular tissue of rates in each group, the positive staining was mainly positioned to the peripheral Sertoli cells in the testicular seminiferous tubule.5. Realtime-PCR detection FSHR mRNA results showed that in comparison of Morinda Officinalis extract lOg/kg group,20g/kg group,30g/kg group and 40g/kg group with CTX+NS group, the expression of FSHR mRNA increased significantly, which was statistically significant (P<0.05), the Morinda Officinalis extract 30g/kg group had the highest expression of FSHR mRNA.6. Results of Realtime-PCR detection of ABP mRNA showed that in comparison of Morinda Officinalis extract lOg/kg group,20g/kg group,30g/kg group and 40g/kg group with CTX+NS group, the expression of FSHR mRNA increased significantly, which was statistically significant(P<0.05).As the concentration of Morinda Officinalis extract increased, the expression of ABP mRNA was also elevated, which was highest in 40g/kg group.7. Western-bolt detection FSHR protein results showed that in comparison of Morinda Officinalis extract lOg/kg group,20g/kg group,30g/kg group and 40g/kg group with CTX+NS group, the expression of FSHR protein increased significantly, which was statistically significant (P<0.05), the Morinda Officinalis extract 30g/kg group had the highest expression of FSHR protein.Conclusion:1. Successfully construct a model of adult male SD rat with spermatogenic function disorders that is treated with intraperitoneal injection of cyclophosphamide.2. Morinda officinalis extract has recovery effects on the spermatogenic function of injured testis of male rat.3. Morinda officinalis extract has improved the expression level in FSHR mRNA and FSHR protein on sustentacular cells.4. Morinda officinalis can promote sustentacular cells to composite and secrete ABP.5. Morinda officinalis, promotes functions of sustentacular cells through improving the FSHR expression on sustentacular cells, so as to recover injured spermatogenic functions of the rats and promote spermatogenesis.