BACKGROUND: Exposure to the ionizing radiation (IR) encountered outside the magnetic field of the Earth poses a persistent threat to the reproductive functions of astronauts. The potential effects of space IR on the circadian rhythms of male reproductive functions have not been well characterized so far. METHODS: Here, we investigated the circadian effects of IR exposure (3 Gy X-rays) on reproductive functional markers in mouse testicular tissue and epididymis at regular intervals over a 24-h day. For each animal, epididymis was tested for sperm motility, and the testis tissue was used for daily sperm production (DSP), testosterone levels, and activities of testicular enzymes (glucose-6-phosphate dehydrogenase (G6PDH), sorbitol dehydrogenase (SDH), lactic dehydrogenase (LDH), and acid phosphatase (ACP)), and the clock genes mRNA expression such as Clock, Bmal1, Ror-α, Ror-β, or Ror-γ. RESULTS: Mice exposed to IR exhibited a disruption in circadian rhythms of reproductive markers, as indicated by decreased sperm motility, increased daily sperm production (DSP), and reduced activities of testis enzymes such as G6PDH, SDH, LDH, and ACP. Moreover, IR exposure also decreased mRNA expression of five clock genes (Clock, Bmal1, Ror-α, Ror-β, or Ror-γ) in testis, with alteration in the rhythm parameters. CONCLUSION These findings suggested potential health effects of IR exposure on reproductive functions of male astronauts, in terms of both the daily overall level as well as the circadian rhythmicity.
ARNTL Transcription Factors/genetics* ; Acid Phosphatase ; Animals ; CLOCK Proteins/genetics* ; Circadian Rhythm/radiation effects* ; Epididymis/radiation effects* ; Gene Expression/radiation effects* ; Genitalia, Male/radiation effects* ; Glucosephosphate Dehydrogenase ; L-Iditol 2-Dehydrogenase ; L-Lactate Dehydrogenase ; Male ; Mice ; Mice, Inbred C57BL ; Models, Animal ; Nuclear Receptor Subfamily 1, Group F, Member 1/genetics* ; Nuclear Receptor Subfamily 1, Group F, Member 2/genetics* ; Nuclear Receptor Subfamily 1, Group F, Member 3/genetics* ; RNA, Messenger/genetics* ; Radiation Exposure ; Radiation, Ionizing ; Reproductive Physiological Phenomena/radiation effects* ; Sperm Motility/radiation effects* ; Spermatozoa/radiation effects* ; Testis/radiation effects*

OBJECTIVE: To investigate the effect of a modified Wuzi Yanzong Pill (, WZYZP) on the male rats' testis after microwave radiation, as well as its potential mechanism. METHODS: Forty-five male rats were randomly assigned to three groups: the control group, the radiation group, and the WZYZP group. The rats in the radiation group and WZYZP group were exposed to microwave radiation for 15 min once, while the rats in the control group were not exposed to any radiation. The rats in the WZYZP group were given a modified of WZYZP by gavage daily for 7 days. Apoptosis in the testis was evaluated using terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) assay. Histopathological alterations of the testis were observed by haematoxylin-eosin (HE) staining. Tat-interactive protein, 60kD (Tip60) and p53 expressions were determined by Western blotting. RESULTS: The apoptosis index (AI) in the radiation group was higher than that of the WZYZP group and control group on day 1 (D1), day 7 (D7) day 14 (D14) after radiation (P<0.05). The seminiferous tubules were of normal morphology in the control group. In the radiation group, the partial seminiferous tubules were collapsed, basement membranes of the seminiferous epithelia became detached. WZYZP could restore the morphological changes. There was no expression of Tip60 among the three groups on D7 and D14. The expression of p53 was higher in the radiation group than in the control group (P<0.05). WZYZP could down-regulate the rising p53 induced by radiation on D7 and D14 (P<0.05). CONCLUSION A modified WZYZP may affect germ cells, and its protective effects may partly result from its ability to intervene in Tip60 mediated apoptosis.
Animals ; Apoptosis ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Male ; Microwaves ; Rats, Wistar ; Testis ; drug effects ; metabolism ; pathology ; radiation effects ; Trans-Activators ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

ObjectiveTo investigate the influence of cellphone electromagnetic radiation (CER) on the testicular ultrastructure and the apoptosis of spermatogenic cells in male rats.atability, feasibility, applicability, and controllability in the construction of experimental animal models, we compared the major anatomic features of the penis of 20 adult beagle dogs with those of 10 adult men. Using microsurgical techniques, we performed cross-transplantation of the penis in the 20 (10 pairs) beagle dogs and observed the survival rate of the transplanted penises by FK506+MMF+MP immune induction. We compared the relevant indexes with those of the 10 cases of microsurgical replantation of the amputated penis.

METHODSThirty adult male SD rats were equally randomized into a 2 h CER, a 4 h CER, and a normal control group, the former two groups exposed to 30 days of 900 MHz CER for 2 and 4 hours a day, respectively, while the latter left untreated. Then the changes in the ultrastructure of the testis tissue were observed under the transmission electron microscope and the apoptosis of the spermatogenic cells was determined by TUNEL.

RESULTSCompared with the normal controls, the rats of the 2 h CER group showed swollen basement membrane of seminiferous tubules, separated tight junction of Sertoli cells, increased cell intervals, apparent vacuoles and medullization in some mitochondria, and increased apoptosis of spermatogenic cells, mainly the apoptosis of primary spermatocytes (P<0.05 ). In comparison with the 2 h CER group, the animals of the 4 h CER group exhibited swollen basement membrane of seminiferous tubules, more separated tight junction of Sertoli cells, wider cell intervals, incomplete membrane of spermatogonial cells, fragments of cytoplasm, nuclear pyknosis and notch, slight dilation of perinuclear space, abnormalities of intracellular mitochondria with vacuoles, fuzzy structure, and fusion or disappearance of some cristae, and increased damage of mitochondria and apoptosis of spermatogenic cells, including the apoptosis of spermatogonial cells, primary spermatocytes, and secondary spermatocytes (P<0.05 ).

CONCLUSIONSCER can damage the testicular ultrastructure and increase the apoptosis of spermatogenic cells of the male rat in a time-dependent manner, and the apoptosis of spermatogenic cells may be associated with the damage to mitochondria.

Animals ; Apoptosis ; Cell Phone ; Electromagnetic Radiation ; Male ; Mitochondria ; radiation effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; radiation effects ; Sertoli Cells ; radiation effects ; Spermatocytes ; radiation effects ; Spermatogonia ; radiation effects ; Testis ; radiation effects ; ultrastructure

OBJECTIVETo investigate the effects of Morina Officinalis How (MOH) on the abnormal levels of serum luteotrophic hormone (LH) and LH receptor (LHR) in the testis tissue induced by cellphone radiation (CPR) in rats.

METHODSFifty adult male SD rats were randomly divided into five groups of equal number: sham CPR, untreated CPR, negative double distilled water (DDW) control, aqueous MOH extract, and alcohol MOH extract. All the animals were exposed to mobile phone radiation except those of the sham CPR group. Then, the rats of the latter two groups were treated intragastrically with MOH at 20 g per kg of the body weight per day in water and alcohol, respectively. After 2. weeks of treatment, all the rats were sacrificed for measurement of the levels of serum LH and LHR in the testis tissue.

RESULTSThe levels of serum LH and LHR were 30.15 ± 8.71 and 33.28 ± 6.61 in the aqueous MOH group and 0.96 ± 0.06 and 0.94 ± 0.08 in the alcohol MOH group, both significantly decreased as compared with the negative DDW controls (P < 0.05), but with no remarkable difference between the two MOH groups (P > 0.05).

CONCLUSIONMOH can improve CPR-induced abnormality of LH and LHR in adult male rats.

Animals ; Cell Phone ; Electromagnetic Radiation ; Luteinizing Hormone ; blood ; drug effects ; radiation effects ; Male ; Morinda ; chemistry ; Radiation Injuries, Experimental ; blood ; drug therapy ; Random Allocation ; Rats ; Receptors, LH ; blood ; drug effects ; radiation effects ; Testis ; radiation effects

OBJECTIVETo observe the effect of Liuweidihuang Pills in relieving cellphone electromagnetic radiation-induced histomorphological abnormality, oxidative injury, and cell apoptosis in the rat testis.

METHODSThirty adult male SD rats were equally randomized into a normal, a radiated, and a Liuweidihuang group, the animals in the latter two groups exposed to electromagnetic radiation of 900 MHz cellphone frequency 4 hours a day for 18 days. Meanwhile, the rats in the Liuweidihuang group were treated with the suspension of Liuweidihuang Pills at 1 ml/100 g body weight and the other rats intragastrically with the equal volume of purified water. Then all the rats were killed for observation of testicular histomorphology by routine HE staining, measurement of testicular malondialdehyde (MDA) and glutathione (GSH) levels by colorimetry, and determination of the expressions of bax and bcl-2 proteins in the testis tissue by immunohistochemistry.

RESULTSCompared with the normal controls, the radiated rats showed obviously loose structure, reduced layers of spermatocytes, and cavitation in the seminiferous tubules. Significant increases were observed in the MDA level (P < 0.01) and bax expression (P < 0.01) but decreases in the GSH level (P < 0.01) and bcl-2 expression (P < 0.01) in the testis issue of the radiated rats. In comparison with the radiated rats, those of the Liuweidihuang group exhibited nearly normal testicular structure, significantly lower MDA level (P < 0.05), bax expression (P < 0.01), and bcl-2 expression (P < 0.01).

CONCLUSIONLiuweidihuang Pills can improve cellphone electromagnetic radiation-induced histomorphological abnormality of the testis tissue and reduce its oxidative damage and cell apoptosis.

Animals ; Apoptosis ; drug effects ; radiation effects ; Body Weight ; drug effects ; radiation effects ; Cell Phone ; Drugs, Chinese Herbal ; pharmacology ; Electromagnetic Radiation ; Glutathione ; metabolism ; Male ; Malondialdehyde ; metabolism ; Oxidative Stress ; Radiation-Protective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; drug effects ; radiation effects ; Spermatocytes ; drug effects ; metabolism ; radiation effects ; Staining and Labeling ; Testis ; drug effects ; metabolism ; pathology ; radiation effects

OBJECTIVETo investigate the effect of Inonotus obliquus polysaccharides on testicular injury induced by exposure to high power microwave (HPM) in rats.

METHODSA total of 30 male Wistar rats were randomly divided into 5 groups, i.e., the normal control group, the microwave radiation model group, the treatment group, the new microwave radiation model group, and the prevention group, 6 in each group. All rats, except those in the normal control group, were exposed to microwave at an average power density of 200 mW/cm2 for 6 min. Rats in the control group and the model group were administered with normal saline by gastrogavage, once a day. Rats in the treatment group and the prevention group were given with Inonotus obliquus polysaccharides by gastrogavage, 2 mL each time (400 mg/kg body weight), once a day. All rats were sacrificed on the 11th day.The sperm density and the rate of sperm deformity were determined. Pathological changes of testis were observed by light microscope and transmission electron microscope.

RESULTSShort-term HPM irradiation could significantly reduce the sperm density and increase the sperm deformity rate (P < 0.05). Meanwhile, obvious pathological changes of testes occurred. Compared with the two model groups, the sperm density increased and the sperm deformity rate decreased in the treatment group and the prevention group (P < 0.05). Under the light microscope, injuries of spermatogenic cells and stromal cells, as well as vascular dilatation and congestion were obviously alleviated in the treatment group and the prevention group. Mitochondrial swelling and endoplasmic reticulum expansion shown by ultrastructural observation were also significantly alleviated. Of them, injuries of spermatogenic cells and inflammation response were milder in the treatment group than in the prevention group.

CONCLUSIONSInonotus obliquus polysaccharides had significant protective effect on microwave radiation induced testicular injury. Better effect was obtained by therapeutic medication than preventive medication.

Animals ; Basidiomycota ; chemistry ; Male ; Microwaves ; adverse effects ; Polysaccharides ; pharmacology ; Radiation Injuries, Experimental ; prevention & control ; Radiation-Protective Agents ; pharmacology ; Rats ; Rats, Wistar ; Testis ; drug effects ; pathology ; radiation effects

OBJECTIVETo investigate the effects of different concentrations of Morinda Officialis How (MOH) extracts on microwave radiation-induced injury to the spermatogenic function of male rats.

METHODSForty SD male rats were equally divided into four groups: control, microwave injury model, aqueous extract of MOH treatment, and alcohol extract of MOH treatment. Models of microwave-induced injury were made by exposing the rats to microwave radiation from a microwave signal generator (900 MHz 1.0 W) at 218 microm/cm2, 12 h/d, for 2 weeks. The model rats of the two treatment groups were intragastrically given aqueous extract and alcohol extract of MOH, respectively, both at 20 g per kg per day for 2 weeks. Then we observed the growth, capture incubation period (CIP), capture times (CT), changes in testicular and epididymal weight and morphology, sperm concentration and malformation, and levels of serum testosterone.

RESULTSCompared with the controls, the rats of the model group showed a slightly reduced body weight, markedly prolonged CIP and decreased CT (P < 0.05), significantly reduced sperm concentration (P < 0.05) and remarkably in- creased sperm malformation (P < 0.05), but no statistically significant differences in the testosterone level. The two treatment groups exhibited obviously decreased body weight, CIP and sperm malformation compared with the control group (P < 0.05) but markedly increased CT, sperm concentration and testosterone level as compared with the models (P < 0.05). The microwave radiation-induced testis injury was repaired perfectly in the two treatment groups, the epididymal ducts filled with sperm and cast-off cells.

CONCLUSIONBoth aqueous and alcohol extracts of MOH can promote spermatogenesis and repair of reproductive injury induced by microwave radiation.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Male ; Microwaves ; adverse effects ; Morinda ; Organ Size ; Plant Extracts ; pharmacology ; Radiation Injuries, Experimental ; drug therapy ; Rats ; Rats, Sprague-Dawley ; Spermatogenesis ; drug effects ; Testis ; drug effects ; Testosterone ; blood

OBJECTIVETo investigate the role of claudin-11, a tight junction component of Sertoli cells, in spermatogenic dysfunction induced by oxidative stress in mice exposed to local radiation.

METHODSWe randomly allocated 48 male Kunming mice to a blank control group (A) and three radiation groups (B, C and D) of equal number, the latter three exposed to local radiation of the lower abdomen with 2 Gy, 6 Gy and 10 Gy of 60Co-gamma-ray, respectively, to induce oxidative stress. Four weeks later, we killed the animals, obtained their body and testis weights, observed the histological changes of the testis by HE staining, measured the levels of serum FSH, testosterone and LH by ELISA, and determined the mRNA levels of claudin-11 and inhibin beta B in Sertoli cells by real time quantitative PCR.

RESULTSAfter exposure to 60Co-gamma-ray radiation, the testis weights were (129.4 +/- 10.81), (87.5 +/- 16.83) and (56.1 +/- 12.36) mg in groups B, C and D, significantly decreased as compared with (182.9 +/- 8.43) mg in group A (P < 0.05); the testis indexes were (3.39 +/- 0.57), (2.46 +/- 0.46) and (1.63 +/- 0.44) mg/g in groups B, C and D, remarkably lower than (4.28 +/- 0.31) mg/g in group A (P < 0.01). Histological analysis revealed obviously decreased diameters of seminiferous tubules, reduced seminiferous epithelia and disarranged spermatogenic cells in the three radiation groups. The tubule differentiation indexes (TDI) were markedly lower in groups B, C and D than in A (P < 0.01). The levels of serum FSH were (6.74 +/- 1.95), (8.41 +/- 2.44) and (10.93 +/- 3.16) IU/L in groups B, C and D, 1.9 times higher in D than in A. With increased dose of radiation, the mRNA levels of inhibin beta in the testis tissue were descended, while the transcription levels of claudin-11 elevated, significantly higher in groups C and D than in A (P < 0.01).

CONCLUSIONLocal radiation-induced testicular oxidative stress can decrease the mRNA level of inhibin beta , increase serum FSH, damage Sertoli cells and elevate the expression of claudin-11 in the testis tissue. Increased claudin-11 and serum FSH may delay the cyclical restitution of hemo-testicular barrier and reduce the number of meiotic spermatocytes in the seminiferous epithelium, which consequently leads to male infertility.

Animals ; Claudins ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Oxidative Stress ; radiation effects ; RNA, Messenger ; genetics ; Seminiferous Tubules ; metabolism ; radiation effects ; Sertoli Cells ; metabolism ; Spermatocytes ; metabolism ; radiation effects ; Spermatogenesis ; Testis ; metabolism ; radiation effects

The study examined the role of endoplasmic reticulum stress (ERS) and signaling pathways of inositol-requiring enzyme-1 (IRE1), RNA-activated protein kinase-like ER kinase (PERK) and activating transcription factor-6 (ATF6) in apoptosis of mouse testicular cells treated with low-dose radiation (LDR). In the dose-dependent experiment, the mice were treated with whole-body X-ray irradiation at different doses (25, 50, 75, 100 or 200 mGy) and sacrificed 12 h later. In the time-dependent experiment, the mice were exposed to 75 mGy X-ray irradiation and killed at different time points (3, 6, 12, 18 or 24 h). Testicular cells were harvested for experiments. H(2)O(2) and NO concentrations, and Ca(2+)-ATPase activity were detected by biochemical assays, the calcium ion concentration ([Ca(2+)]i) by flow cytometry using fluo-3 probe, and GRP78 mRNA and protein expressions by quantitative real-time RT-PCR (qRT-PCR) and Western blotting, respectively. The mRNA expressions of S-XBP1, JNK, caspase-12 and CHOP were measured by qRT-PCR, and the protein expressions of IRE1α, S-XBP1, p-PERK, p-eIF2α, ATF6 p50, p-JNK, pro-caspase-12, cleaved caspase-12 and CHOP by Western blotting. The results showed that the concentrations of H2O2 and NO, the mRNA expressions of GRP78, S-XBP1, JNK, caspase-12 and CHOP, and the protein expressions of GRP78, S-XBP1, IRE1α, p-PERK, p-eIF2α, ATF6 p50, p-JNK, pro-caspase-12, cleaved caspase-12 and CHOP were significantly increased in a time- and dose-dependent manner after LDR. But the [Ca(2+)]i and Ca(2+)-ATPase activities were significantly decreased in a time- and dose-dependent manner. It was concluded that the ERS, regulated by IRE1, PERK and ATF6 pathways, is involved in the apoptosis of testicular cells in LDR mice, which is associated with ERS-apoptotic signaling molecules of JNK, caspase-12 and CHOP.
Animals ; Apoptosis ; physiology ; radiation effects ; Endoplasmic Reticulum Stress ; physiology ; radiation effects ; Male ; Mice ; Radiation ; Testis ; physiology ; radiation effects

OBJECTIVETo investigate the effect of simulated microgravity and carbon ion irradiation (CIR) on spermatogenic cell apoptosis and sperm DNA damage to the testis of male Swiss Webster mice, and assess the risk associated with space environment.

METHODSSperm DNA damage indicated by DNA fragmentation index (DFI) and high DNA stainability (HDS) was measured by sperm chromatin structure assay (SCSA). Apoptosis of spermatogenic cells was detected by annexin V-propidium iodide assay. Bax (the expression levels of p53) and proliferating cell nuclear antigen (PCNA) were measured by immunoblotting; p53 and PCNA were located by immunohistology.

RESULTSHDS, DFI, apoptosis index, and the expression levels of p53 and Bax were detected to be significantly higher in the experimental groups (P<0.05) compared with those in the control group; however, the PCNA expression varied to a certain degree. p53- and PCNA- positive expression were detected in each group, mainly in relation to the spermatogonic cells and spermatocytes.

CONCLUSIONThe findings of the present study demonstrated that simulated microgravity and CIR can induce spermatogenic cell apoptosis and sperm DNA damage. Sperm DNA damage may be one of the underlying mechanisms behind male fertility decline under space environment. These findings may provide a scientific basis for protecting astronauts and space traveler's health and safety.

Animals ; Apoptosis ; radiation effects ; Carbon ; Cell Proliferation ; radiation effects ; DNA Damage ; Heavy Ions ; adverse effects ; Immunohistochemistry ; Male ; Mice ; Random Allocation ; Sperm Count ; Spermatogenesis ; radiation effects ; Spermatozoa ; radiation effects ; Testis ; radiation effects ; Weightlessness Simulation