| With the development of human society, mankind is faced with more than the risk of natural background radiation; nearly half a century to the world male fertility declined significantly; testicular tissue as a target organ sensitivity to ionizing radiation, ionizing radiation has been become a cause of human male sexual dysfunction and infertility is one of the most common cause; male reproductive health is facing a very serious situation, improvement and maintenance of male reproductive health have become an increasing concern.Laminaria japonica polysaccharides is a kind of natural macromolecular substances whitch exists in Intercellular and Intracellular of laminaria japonica, and include three kinds of elements:Algin, Laminarin and Fucoidan.Studies have shown that Laminaria japonica polysaccharides is not only anti-coagulation, anti-arteriosclerosis, anti-tumor, anti-virus and other biological activity, also has anti-radiation; but in several small doses of local radiation and the field of reproductive function protection system has rarely been reported.Therefore, this research was to study the affects of repeated small doses of local radiation to the o sexual function, spermatogenic function, testicular cell apoptosis and related molecules of male rats; to study the potential mechanisms of the Interfere with recovery of reproductive dysfunction repeated small doses of laminarin on the local radiation-induced reproductive injury may interfere with recovery mechanisms, and from the overall level of cells and animals many small doses of laminarin on the local radiation-induced damage of testicular cells and its molecular mechanism. To provide a scientific basis for the mechanisms of anti-radiation by UP, and to provide theoretical support for the deep processing of LJP and the development of safe and effective eproductive health products.Part I The investigation of mechanism of laminaria japonica polysaccharides interfere the damage of sexual function lead bychronic ionizing radiationObjective:To set up foundation for further study by observing the self recovery and the recovery after interfered by LJP of the sexual function of male rat from damage of chronic ionizing radiation and discussing the potential mechanism. Method:56 Wistar male rats are divided at random into control group,3 model groups and 3 LJP groups after 7d adaptable feeding.3 LJP groups are lavaged with LJP100mg·kg-1·d-1 and other groups with saline. Each group is partly irradiated by 60Coγradial once per day but control group, which last for five continuous times per week and total 20 times with total dose 2.3 Gy for each rat. Erection and mate experiments are performed at 1,7 and 14 day from ceasing irradiation. One day later the rats are executed to collect blood, separate testicle and test indicator of rats'organization, sex gland quality and organ coefficient, mating and erection and LH,FSH, Ts and E2. Result:after chronic ionizing radiation, the model group 1,7, 14d rat's weight and its gland weight gradually increase but testicles weight along with recovery time prolonging. Yet model group rats'weight and gland weight is under control group while LJP group is above model group including testicles weight. Moreover, LJP7, 14d group rats'testicle weight is obviously increased compared to corresponding mode group. Coefficient of model group rats'organs is reduced along with time prolonging, especially testicle. While LJP group coefficient is closer to control group. Model group erection latent period is obviously prolonged and ejaculation frequency is obviously reduced while LJP groups' erection latent period is shortened and ejaculation frequency is increased along with recover time prolonging. Indicator of LJP14d group is closer to control group. Compare with control group, model group 1,7,14d serum LH, FSH, Ts content reduced and declined obviously after ceasing radiation 14d later. Serum E2 is continuously increased yet still under control group. While LJP 1,7,14d serum Ts increased and LH, FSH, E2 declined to be closed to control group. Conclusion:LJP has favorable effect on protection of male rats sexual function which is damaged by chronic ionizing radiation to promote rats weight and its testicle weight that is damaged by radiation, to elevate the coefficient of testicle, to shorten erection latent period, to enhance ejection and to restore serum sex hormone LH, FSH, Ts, and E2; to shorten erection and riding latent period by the regulation of hypothalamus-pituitary gland-goand shafting. LJP can improve sex function obviously.Part II The investigation of mechanism of laminaria japonica polysaccharides interfere the damage of spermatogenous function of testis on rat lead by chronic ionizing radiationObjective:To set up foundation for further study by observing the self recovery and the recovery after interfered by LJP of the sexual function of male rat from damage of chronic ionizing radiation and discussing the potential mechanism. Method:56 Wistar male rats are divided at random into control group, 3 model groups and 3 LJP groups after 7d adaptable feeding. 3 LJP groups are lavaged with LJP100mg·kg-1·d-1 and other groups with saline. Each group is partly irradiated by 60Coγradial once per day but control group, which last for five continuous times per week and total 20 times with total dose 2.3 Gy for each rat. Erection and mate experiments are performed at 1,7 and 14 day from ceasing irradiation. One day later the rats are executed to collect blood, separate testicle and test germ cell and indictor of MDA, GSH, SOD, GSH-PX and LDH. Result:After multiple mini-doses ionizing radiation, the number of spermatozoa and its survival rate are decreased obviously in LJP1d group, modle group and LJP groups compared to control group; but the drcrease trend of spermatozoa in LJP groups is slow down, the survival rate of spermatozoa is increased, there is significiant difference compared with modle group. Observation by optical microscope shows that the damage of sperm tube aggravates along with recovery time prolonging and testicle self repair unobviously. Although LJP groups have the same result, the damage of LJP groups is comparatively mild. Observation it s ultra-structure by electron microscope, we can find that damages are existed in nucleus membrane and Organelles of spermatogenous cell and spermiocytes in modle group, lipid granule increased, the damage is obviously in acrosome and arrhenokaryon of spermatozoa, although the gamage of sertoli cell and spermatid is mild, but there are deformed organelles and lipid granule in cytoplasm. The damage trend of all kinds of testis cells is the same in LJP group to modle group, but the lipid granule in in cytoplasm is more fewer than that of in modle group, the damage degree of organelles and spermatozoa is less than that of in modle group, and the number of spermatozoa is more than in modle group. After cessation of radiation, the vitality of SOD and GSH-PX in testis tissue of rat is obviously decreased in modle group, and there is significant difference between the content of MDA and GSH compared with control group, But in LJP groups, the content of MDA is decreased, there is no significant difference of the content of MDA in LJP 14d group, the content of GSH and the vitality of SOD,GSH-PX are increased compared with modle 14d group with the arrest of radiation time prolonging. The vitality of LDH is continuing decreasing in modle groups, and the difference is significant compared with control group, but it is almost the same with control group in testis tissue of LJP groups. Conclusion:LJP has satisfactory promotive effect to spermatogenic dysfunction of testis tissue on male rat after chronic ionizing radiation, the mechanism is likely to decresses the content of MDA, increases GSH in testis tissue, improves the vitality of SOD,GSH-PX and LDH in testis tissue, promote the recovery of anti-oxidant system in testis, improves it's spermatozoa count and survival rate,relieve the damage of convoluted tubule, maintains the better morphous of androgone and it's organelles in testis, and promotrs the recovery of spermatogenic dysfunction of testis on male rat obviously.Part III The investigation of mechanism of laminaria japonica polysaccharides interfere the recovery of testicle cells injury on rat lead by chronic ionizing radiationObjective:To set up foundation for further study by observing the self recovery of apoptosis in testicular cells and the recovery after interfered by LJP of male rat from damage of chronic ionizing radiation and discussing the potential mechanism. Method:56 Wistar male rats are divided at random into control group, 3 model groups and 3 LJP groups after 7d adaptable feeding.3 LJP groups are lavaged with LJP100mg·kg-1·d-1 and other groups with saline. Each group is partly irradiated by Coy radial once per day but control group, which last for five continuous times per week and total 20 times with total dose 2.3 Gy for each rat. the rats are executed at 1,7 and 14 day from ceasing irradiation to separate testicle tissue, make testicle cell suspension, electron microscopic and frozen section of testicle tissue, observe ultra-structure of Mitochondrion in testicle cell, measure the content of testicle cell,membrane potentials of Mitochondrion and density of Ca2+ etal in testicle cell of rat. Result:After cessation of radiation, the apoptosis count of modle group is increasing significantly,spermatozoa and spermatozoa cells are decreasing obviously, in LJP1,7,14d groups, the Percentage of cells in testicular cells in HC peak is obviously lower than that of corresponding modle group; in LJP14d group, the Percentage of cells in testicular cells in 1C,2C,4C peak is much higher than that of in modle14d group. After recovery for 14 days, chronic ionizing radiation to the repair of the mitochondrion s size and structure damage in androgone and spermatozoa of testicle is unobviously; after interfere with LJP, the restoration of mitochondrion is obvious than that of in modle groupand the condition is better. TheΔΨm of mitochondrion in modle groups is continuely decreases than that of in control groups after radiation damage; therefore, after interfere with LJP, theΔΨm of mitochondrion is obviously higher than that of in modle groups. The density of Ca2+ in testicle cell of modle groups is significantly higher than that of in control groups; but it is obviously lower in LJP 7 and LJP 14 groups than that of in corresponding modle groups. Conclusion:LJP has depressant effect to apoptosis of testicle cell on male rat which lead by multiple chronic ionizing radiation, the mechanism is likely to cuts down the count of apoptosis in testis, improves the ultra-structure of sperm tail and mitochondrion of testicle cell, elevates the membrane potentials of mitochondrion, reduces the density of Ca2+ in testicle cell, promotes self recovery of damaged testicle cell, avoides abundant generation of apoptosis cells.Part IV The investigation of mechanism of laminaria japonica polysaccharides interfere the molecule of testicle cell on rat lead by chronic ionizing radiationObjective:To observe the regulation of abnormality expression of molecular in testicular cells damaged by chronic ionizing radiation to the injury of reproductive function on male rat and the regulation effect after interfered with LJP, to discuss the molecular mechanism of LJP to the recovery of reproductive function injury on rat. Method:56 Wistar male rats are divided at random into control group,3 model groups and 3 LJP groups after 7d adaptable feeding. 3 LJP groups are lavaged with LJP100mg·kg-1·d-1 and other groups with saline. Each group is partly irradiated by 60Coγradial once per day but control group, which last for five continuous times per week and total 20 times with total dose 2.3 Gy for each rat. the rats are executed at 1,7 and 14 day from ceasing irradiation,separates the tisticke tissue of rat, determines the damage of DNA in testicle cell, the differential expression of P53 and Hsp70 protein and gene in testicle cell and the expression of Bcl-2/Bax, Fas and casapse3 in testicle tissue. Result:The comets tailing rate and the tail length of testicle cell in modleld, modle 7d and modle 14d group is aggravate with the recovery time prolonging after radiation, and there is significant difference compared to the control group; although the comets tailing rate and the tail length of testicle cell in LJP groups is also aggravate with the recovery time prolonging, but is obviously palliate than that of in corresponding modle groups. Bcl-2 decreasing but Bax increasing significantly in modle groups compared to control groups, the ratio of Bcl-2 and Bax is depressed; after recovery for 14 days, Fas, caspase3 and P53 are increasing but Hsp70 is decreasing obviously. The change tendency of P53 in LJP groups is the same to modle groups,although P53 in LJP14d group is notable increasing compared to control group, but is decreased obviously than that of in corresponding modle14d group. Bcl-2 in LJP groups is notable lower than that of in control groups, but the tendency is rosen; and in LJP14d group, Bcl-2 is increasing than that of in corresponding modle group; Bax is also shows a set-up tendency, but in LJP groups is notable lower than that of in corresponding modle groups; Bcl-2/Bax is downtrend, but the ratio is greater than modle groups. Alougth the expression of Fas in LJP groups is also shows a set-up tendency, but are all obviously lower than those of in corresponding modle groups. The Caspase3 in LJP groups is increasing continuely, higher than that of in control groups significantly, but is decreasing compared to corresponding modle groups. The expression of Hsp70 in LJP 1d group is notable higher than that of in control group, but is decreasing significantly in LJP 14d group compared to control group, and the content of Hsp70 in LJP 14d group is increasing obviously than that of in modle 14d group. Conclusion:LJP has apparent regulation effect to the molecular expression of testicle cell on male rat lead by chronic ionizing radiation, induces depression of the expression of P53 in testicle tissue on rat after 14 days recovery, lead to the reduction of Bax s expression, elevation of Bcl-2, aubmentation of Bcl-2/Bax; the expression of Hsp70 is elevated, the expression of Fas is reduced, so the Caspase-3 is decreased, and it promotes the damage repair of DNA in testicle cell, abates the cell apoptosis, protects the germ cell of rat, and improves it s reproductive function.
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