| Objective：To establish microwave induced reproductive impairment animal model of adult sdmale rat, then to investigate the effect on the male rats’ reproductive and ifhypothalamus promote spermatogenesis after given Radix Morinda Officialis extract,in addiation, to detect the levels of some biochemical criterion. Eventually, to providethe theoretical bases of effective component screenin from Radix Morinda Officialisand research on active monomer.Methods：75sd male rats were average divided into there groups as blank control group (5)sham exposed control group (35) and exposed group (35). We have been to draw thematerials from5rats in each group. And then, the rats of exposed group wererandomly divided into there groups as microwave model group, Radix MorindaOfficialis aqueous extract treatment group and Radix Morinda Officialis alcoholextract treatment group after exposure,10for each. At the same time, sham exposedcontrol group also were average randomly divided into three groups as control group,Radix Morinda Officialis aqueous extract given group and Radix Morinda Officialisalcohol extract given group.The rats of exposed group were given microwaveradiation with Microwave Signal Generator (900MHz1.0W),218μm/cm2,12h/d,for2weeks. And then Radix Morinda Officialis aqueous extract treatment group ratswere given Radix Morinda Officialis aqueous extract (20g/kg.d-1,for2weeks), thesame to Radix Morinda Officialis aqueous extract given group. Radix MorindaOfficialis alcohol extract treatment group rats were given Radix Morinda Officialisalcohol extract (20g/kg.d-1,for2weeks), the same to Radix Morinda Officialis alcoholextract given group.The growth of rats, the sexual function: capture incubation period (CIP) and capture times(CT), the morphological changes on testes, epididymis andhypothalamus, testis-weight ratio and epididymis-weight ration, sperm concentration，malformed sperm ratio, serum hormone (GnRH, LH, FSH, T) levels and theexpression of GnRH-mRNA on hypothalamus arcuate nucleus and Median Eminencewere assessed.Results：1. There were no statistically significant differences between exposed group (255±27) g and sham exposed control group (265±20) g on body weight. But the bodyweight instinctly decreased after given Radix Morindae Officinalis compared withblank control group. The rats of microwave model group showed that yellowmousefur, dry rot and reluster, especially significant in caudal1/3place comparedwith control group, and to become more and more serious along with extendingradiation time. The performance of neuroethology in rats from daytime sleep andthe night awake, physical agility, defense attack ability to restlessness, irritabilityin the process of exposure. Compared with microwave model group, damagedchanges on the rats in two treatment groups slowly be reduced with regulartreatment after given different extract of Radix Morindae Officinalis. There wasno significant difference between the treatment groups and the control group onthe surface characteristics, but we found that the therapeutic effects of aqueousextract of Radix Morindae Officinalis treatment group was better than alcoholextract of Radix Morindae Officinalis.2. The sexual function include capture incubation period (CIP) and capturetimes(CT). Compared with sham exposed group, CIP was significantly increasedand CT was instinctly decreased in microwave model group (P<0.05).3. Testis-weight ration and Epididymidis-weight ration were significantly decreasedcompared with other groups but Radix Morindae Officinalis alcohol extracttreatment group (P<0.05). 4. Histological assessments were observed with a light microscope afterHematoxylin-eosin (HE) staining. In blank control group, seminiferous epitheliumof testis to line up in order, the level of androgone, spermatocytes and spermatiddistribution well-arranged. A great quantity sperm was seen in seminiferoustubule and epididymal duct. In microwave model group, the germ cells ofgerminal epithelium irregulation and deformation in the spermatogenesis,spermatogenic cells arrangement disorder, fall off, necrosis. Cast-off cells andnecrotic cellular debris were filled with seminiferous tubules or tube cavity empty.Only we can see a small quantity sperm in seminiferous tubules or filled withnothing. Sometimes, we may see some red blood cells (RBC) freeing outsideseminiferous tubule. interstitial cells scattered distribution. There are threechanges in epididymis tube on microwave model group:①the epididymis tubefilled with cast-off cells and necrotic cellular debris;②the epididymis tube filledwith cast-off cells, necrotic cellular debris and a small amount of sperm;③theepididymis tube emptiness. In Radix Morindae Officinalis aqueous extracttreatment group, seminiferous tubule cells and new sperms (namely sperm fromthe basal part trending intraluminal) in seminiferous tubule were increased, mostof the epididymis tube are full of sperm, occasional1~2empty pipe cavity exist.In Radix Morindae Officinalis alcohol extract treatment group still visible mostspermatogenic cells necrosis and desquamate, sperm rarely. Epididymis tubeexfoliated cells and cell debris sparse distribution, emptiness bureaucratic tone isstill visible.Ultrastructure assessments were observed with transmission electron microscopy(TEM). Spermatogenesis cells were arrayed closely in control group and also alarge number of sperms can be seen. In microwave model group, large areas ofnecrosis in testis, in sertoli cells and spermatogenic cells, the mitochondria swellingand vacuolar degeneration, nuclear fission anomaly as the central empty ring andvisible nuclei dissolved, visible very small amount of sperms. In Radix MorindaeOfficinalis aqueous extract treatment group, sperm count significantly increased inseminiferous tubule. In Radix Morindae Officinalis alcohol extract treatment group that spermatogenic cells recovery significantly but have no seen sperm countincreased.5. Sperm parameters include sperm concentration and sperm malformed ratio wereevaluated by sperm count. sperm concentration of microwave model group wassignificantly decreased and sperm malformed ratio was instinctly increasedcompared with other groups (P<0.05, respectively) but Radix MorindaeOfficinalis alcohol extract treatment group.6. The level of testosterone in microwave model group was decreased significantlycompared with blank control group, also gonadotropin-releasing hormone leveland luteinizing hormone level were increased apparently. There was nostatistically significant differences in GnRH, FSH, LH and T between RadixMorindae Officinalis alcohol extract treatment group and microwave modelgroup.7. Ultrastructure of hypothalamus was assessed by TEM. The neuro cells weredamaged distinctly in microwave model group.It showes that the mitochondriaswelling and vacuolar degeneration, Endoplasmic reticulum expansion,myelinated nerve fiber demyelination, axon swelling. The damage structures weredecreased in Radix Morindae Officinalis extract treatment group.8. GnRH main expression in arcuate nucleus and expression enhanced in microwavemodel group. The results were found by Immunohistochemisty (IHC)andImmunofluorescence (IF).9. Real-time PCR indicate that GnRH-mRNA in microwave model group expresssignificantly.Conclusions：1. Reproductive impairment animal model of adult sd male rat was successful builedby microwave radiation for2weeks.2. Radix Morinda Officialis aqueous extract can promote spermatogenesis andsperm maturation in testis and encourage the plerosis of injured reproductive organs. But there is unconspicuous about the effect Radix Morinda Officialisalcohol extract to spermatogenesis.3. Microwave radiation on hypothalamus has a certain degress of damage, MorindaOfficinalis How against the expression of GnRH in hypothalamus.4. The effect of Radix Morinda Officialis aqueous extract on reproductive system isbetter than Radix Morinda Officialis alcohol extract effection. |
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