ObjectiveTo study the effect of Erxian Decoction (EXD) on oligospermia (OS) induced by cyclophosphamide in mice.

METHODSEighty 6-week-old male Kunming mice were randomly divided into five groups of equal number, normal control, OS model control, and low-, medium- and high-dose EXD, the former two groups treated intragastrically with normal saline and the latter three with EXD at 3, 6 and 12 g per kg of the body weight qd for 30 days. From the 21st day of administration, the mice of the normal control group were injected intraperitoneally with saline and those of the other four groups with cyclophosphamide at 80 mg per kg of the body weight qd for 5 consecutive days. At 24 hours after the last gavage, the bilateral epididymides of the mice were collected and sperm suspension prepared for determination of the sperm count and motility, and the bilateral testes were harvested for histomorphological observation and measurement of the concentrations of superoxide dismutase (SOD), malondialdehyde (MAD) and glutathione (GSH) in the testis tissue.

RESULTSCompared with the normal controls, the mice of the OS model control group showed significant decreases in epididymal sperm concentration (［9.31 ± 1.32］ vs ［3.32 ± 1.13］×107/ml, P <0.01) and motility (［44.75 ± 8.12］% vs ［25.95 ± 11.41］， P<0.01) and the concentrations of SOD (［37.27 ± 0.99］ vs ［14.23 ± 1.99］ U/mg prot, P <0.01) and GSH (［101.55 ± 8.74］ vs ［58.77 ± 8.93］ μmol/L, P <0.01) but an obvious increase in the MDA level (［2.21 ± 0.65］ vs ［2.61 ± 0.15］ nmol/mg prot, P <0.05) in the testis tissue. In comparison with the OS model controls, the mice treated with low-, medium- and high-dose EXD exhibited significantly increased epididymal sperm concentration (［8.34 ± 2.59］, ［8.59 ± 1.10］ and ［8.41 ± 1.47］×107/ml) (P <0.01) and motility (［36.04 ± 12.33］%, ［38.87 ± 13.13］% and ［41.90 ± 8.09］%) (P <0.01) and concentrations of SOD (［22.99 ± 1.11］, ［20.82 ± 1.81］ and ［21.33 ± 1.66］ U/mg prot) (P <0.01) and GSH (［104.74 ± 2.47］, ［98.61 ± 12.98］ and ［108.89 ± 5.85］ μmol/L) (P <0.01) but decreased level of MDA (P <0.05).

CONCLUSIONSErxian Decoction can improve cyclophosphamide-induced reduction of sperm concentration and motility, which might be associated with its abilities of resisting oxidation and reducing oxidative stress injury.

Animals ; Cyclophosphamide ; Drugs, Chinese Herbal ; pharmacology ; Epididymis ; Glutathione ; analysis ; Male ; Malondialdehyde ; analysis ; Mice ; Oligospermia ; chemically induced ; drug therapy ; Oxidative Stress ; Random Allocation ; Sperm Count ; Sperm Motility ; drug effects ; physiology ; Spermatozoa ; drug effects ; Superoxide Dismutase ; analysis ; Testis ; anatomy & histology ; chemistry ; drug effects

ObjectiveTo investigate the effects of resveratrol in the cryopreservation medium on the quality and function of post-thaw sperm.

METHODSSemen samples were obtained from 50 normozoospermic and 50 oligoasthenozoospermic men, liquefied and then cryopreserved in the glycerol-egg yolk-citrate (GEYC) medium with or without 30 μmol/L resveratrol. Sperm motility, viability and acrosome reaction (AR) were examined before and after thawing. Sperm lipid peroxidation and the level of reactive oxygen species (ROS) were measured using commercial malondialdehyde (MDA) and the ROS assay kit. Sperm mitochondrial membrane potential (MMP) and DNA damage were determined by Rhodamine 123 staining and TUNEL.

RESULTSThe percentage of progressively motile sperm (PMS), total sperm motility, sperm viability, MMP and AR were significantly decreased (P <0.05) while the levels of sperm ROS, MDA and DNA fragmentation index (DFI) remarkably increased in both the normozoospermia and oligoasthenozoospermia groups after cryopreservation as compared with those in the fresh ejaculate (P <0.05). In comparison with the non-resveratrol control, the post-thaw sperm cryopreserved with 30 μmol/L resveratrol showed markedly higher PMS (［32.7 ± 4.8］ vs ［43.1 ± 6.3］ %, P <0.05), total motility (［44.8 ± 6.9］ vs ［56.9 ± 7.4］ %, P <0.05), viability (［52.3 ± 6.1］ vs ［67.5 ± 5.6］ %, P <0.05), MMP (［56.5 ± 7.0］ vs ［63.4 ± 7.5］ %, P <0.05) and AR (［16.6 ± 3.8］ vs ［26.3 ± 4.7］ %, P <0.05) but lower ROS, MDA and DFI (all P <0.05) in the normozoospermia group, and so did the post-thaw sperm in the oligoasthenozoospermia group, with a particularly lower DFI (［28.5 ± 4.8］ vs ［36.3 ± 5.7］%, P <0.01).

CONCLUSIONSResveratrol in the cryopreservation medium can improve the quality and function of post-thaw human sperm by reducing cryopreservation-induced sperm injury and the level of ROS.

Acrosome ; drug effects ; Animals ; Antioxidants ; pharmacology ; Cryopreservation ; methods ; DNA Fragmentation ; Humans ; Lipid Peroxidation ; Male ; Malondialdehyde ; Membrane Potential, Mitochondrial ; Reactive Oxygen Species ; analysis ; Resveratrol ; pharmacology ; Semen Analysis ; Semen Preservation ; adverse effects ; Sperm Motility ; drug effects ; Spermatozoa ; drug effects ; physiology

ObjectiveTo explore the antagonistic effect of vitamin E (VE) on male reproductive toxicity induced by di-2-ethylhexyl phthalate (DEHP) in pubertal SD rats and its underlying mechanisms.

METHODSThirty 5-week-old male SD rats were randomly divided into five groups of equal number, corn oil control, low-dose (10 mg/kg/d), medium-dose (100 mg/kg/d) and high-dose DEHP exposure (500 mg/kg/d), and VE intervention (high-dose DEHP + VE ［100 mg/kg/d］), and treated respectively for 30 successive days. At 3 days after treatment, the testes of the animals were harvested for determination of the oxidative stress index, serum reproductive hormone levels, cauda epididymal sperm parameters, and expressions of cell apoptosis-related genes and proteins.

RESULTSCompared with the control group, the rats of the medium- and high-dose DEHP groups showed significant decreases in the levels of such serum reproductive hormones as follicle-stimulating hormone (FSH), luteinizing hormone (LH) and testosterone (T), sperm parameters as average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), straightness (STR), linearity (LIN) and wobble (WOB), and the activities of superoxide dismutase (SOD) and glutathione peroxide (GSH-Px), but significant increases were observed in the latter two groups in the content of malondialdehyde (MDA)(［3.32±0.87］ nmol/mg pro vs ［2.13±0.49］ nmol/ mg pro), mRNA expressions of Bad, Bax, Cytochrome C, Caspase-3 and the Bax/Bcl-2 ratio, and protein expressions of Cytochrome C and Caspase-3. In comparison with the high-dose DEHP group, the VE intervention group exhibited remarkably increased serum LH and T levels, sperm VAP, VSL, VCL, STR and WOB, and activities of SOD and GSH-Px, but markedly decreased mRNA expressions of Bad, Bax, Cytochrome C, Caspase-3 and the Bax/Bcl-2 ratio as well as the protein expressions of Cytochrome C and Caspase-3 in the testis tissue (P<0.05).

CONCLUSIONSExposure to DEHP induces androgen secretion disorders, causes oxidative damage to the testicular tissue, activates the mitochondrial apoptosis pathway in the testis, and ultimately reduces the quality of epididymal sperm, while VE can protect the rat testis from DEHP-induced reproductive toxicity.

Animals ; Antioxidants ; pharmacology ; Apoptosis ; genetics ; Autophagy-Related Protein 5 ; metabolism ; Caspase 3 ; metabolism ; Diethylhexyl Phthalate ; antagonists & inhibitors ; Epididymis ; Follicle Stimulating Hormone ; blood ; Luteinizing Hormone ; blood ; Male ; Malondialdehyde ; metabolism ; Mitochondria ; drug effects ; Oxidative Stress ; drug effects ; Oxidoreductases ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reproduction ; Spermatozoa ; drug effects ; physiology ; Superoxide Dismutase ; metabolism ; Testis ; drug effects ; Testosterone ; blood ; Vitamin E ; pharmacology

Objective: To observe the effect of Huangjing Zanyu Capsule (HZC) on sperm mitochondrial membrane potential (MMP) in asthenozoospermia patients. METHODS: We assigned 70 asthenozoospermia patients to a treatment group (n = 39) and a control group (n = 31), the former treated with oral HZC at the dose of 4 capsules tid for 3 months while the latter left untreated. We obtained semen parameters from the patients and detected their sperm mitochondrial membrane potentials (MMP) by JC-1 staining and flow cytometry before and after medication, followed by comparison between the two groups. RESULTS: The total effectiveness rate was 71.05% in the treatment group and natural pregnancy was achieved in 3 cases during the medication. A total of 35 patients in the treatment group and 30 controls completed all the laboratory examinations after a 3-month observation. Compared with the controls, the patients treated with HZC exhibited significant improvement after medication in MMP (variation value: ［1.19 ± 10.36］% vs ［20.28 ± 14.21］%, P <0.01), total sperm motility (variation value: ［3.46 ± 8.67］% vs ［20.68 ± 14.12］%, P <0.01), the percentage of progressively motile sperm (variation value: ［2.26 ± 8.29］% vs ［17.58 ± 12.73］%, P <0.01), and the percentage of morphologically normal sperm (variation value: ［0.23 ± 3.48］% vs ［3.37 ± 3.99］%, P <0.01). MMP was significantly correlated with total sperm motility (r = 0.69, P <0.01), progressive sperm motility (r = 0.75, P <0.01) and normal sperm morphology (r = 0.26, P <0.01). CONCLUSIONS Huangjing Zanyu Capsule can enhance sperm mitochondrial membrane potential and sperm mitochondrial function, thus improving total sperm motility, progressive sperm motility and normal sperm morphology. It is safe and effective for the treatment of asthenospermia.
Asthenozoospermia ; drug therapy ; Capsules ; Case-Control Studies ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; Flow Cytometry ; Humans ; Male ; Membrane Potential, Mitochondrial ; drug effects ; physiology ; Pregnancy ; Semen ; drug effects ; Sperm Motility ; drug effects ; Spermatozoa ; drug effects ; ultrastructure ; Staining and Labeling

Objective: To clarify the roles of yam polysaccharide (YPS) in improving sperm viability and protecting sperm DNA integrity in vitro and provide a new approach to the treatment of oligoasthenozoospermia. METHODS: We collected samples by masturbation from 36 normal fertile males aged 27－39 years. Each sample was divided into six groups: blank control or treated with normal saline, vitamin C solution, and YPS solution at low (0.25 mg/ml), medium (1.0 mg/ml) or high concentration (5.0 mg/ml). Using eosin-Y staining, sperm hypotonic swelling (HOS) and sperm chromatin diffusion (SCD) test, we observed the effects of different concentrations of YPS on sperm viability, membrane integrity and nuclear DNA. RESULTS: After 24 and 48 hours of treatment, sperm viability was markedly reduced in the vitamin C (［28.5 ± 3.1］ and ［6.5 ± 1.2］%), low-YPS (［31.3 ± 3.5］ and ［6.5 ± 2.2］%), medium-YPS (［37.1 ± 3.5］ and ［9.5 ± 2.8］%) and high-YPS groups (［38.3 ± 3.3］ and ［9.0 ± 3.2］%) as compared with the blank control (［17.3 ± 2.1］ and ［3.2 ± 1.3］%) (P <0.01) and normal saline groups (［13.4 ± 4.1］ and ［3.1 ± 2.0］%) (P <0.01), and it was significantly higher in the medium- and high-YPS than in the vitamin C group (P <0.05 and P <0.01). The rate of sperm DNA fragmentation was remarkably decreased at 48 hours in the vitamin C (［30.5 ± 3.1］%), low-YPS (［29.4 ± 2.6］%), medium-YPS (［28.5 ± 2.3］%) and high-YPS groups (［27.9 ± 1.9］%) in comparison with the blank control (［41.7 ± 2.2］%) (P <0.01) and normal saline groups (［42.1 ± 3.3］%), markedly lower in the medium- and high-YPS than in the blank control, normal saline and vitamin C groups (P <0.05 or P <0.01), but with no statistically significant difference between the low-YPS and vitamin C groups (P >0.05). CONCLUSIONS Yam polysaccharide can improve sperm viability and protect sperm DNA integrity in vitro.
Adult ; Ascorbic Acid ; pharmacology ; DNA ; drug effects ; DNA Fragmentation ; Dioscorea ; chemistry ; Humans ; Male ; Polysaccharides ; pharmacology ; Semen Analysis ; Sperm Motility ; Spermatozoa ; drug effects ; physiology ; Vitamins ; pharmacology

Objective: To explore the effects of Zhibai Dihuang Decoction (ZDD) on mitochondrial cytochrome oxidase (COX) in the spermatogenic cells of rats with ureaplasma urealyticum (UU) infection. METHODS: From forty 4－5 months old SD rats, 30 were randomly selected for the establishment of the model of testicular UU infection by inoculating the bladder with UU suspension and the other 10 injected with normal saline as controls (group A). At 7 days after inoculation, the rat models of testicular UU infection were treated orally with normal saline (group B), ZDD at 1 g per kg of the body weight per day (group C), and azithromycin at 0.105 g per kg of the body weight per day (group D), respectively, once daily for 21 days. Then all the animals were sacrificed and the epididymal and testicular tissues collected for examination of sperm motility with the color sperm dynamic detection system, measurement of the COX activity with the immunohistochemical DAB method, and determination of the mRNA expressions of COXⅠ and COXⅡ by RT-PCR. RESULTS: Compared with group A, group B showed significant decreases in such sperm parameters as grade a sperm (［1.03 ± 0.09］ vs ［0.07 ± 0.03］ %, P<0.01), grade b sperm (［2.07 ± 0.52］ vs ［0.35 ± 0.13］ %, P<0.01), straight line velocity (VSL) (［10.95 ± 0.98］ vs ［6.78 ± 1.05］ μm/s, P<0.01), curvilinear velocity (VCL) (［42.03 ± 1.35］ vs ［38.10 ± 7.65］ μm/s, P>0.05), average path velocity (VAP) (［16.22 ± 1.52］ vs ［10.05 ± 1.80］ μm/s, P<0.01), and the mRNA expressions of COX Ⅰ (［2.25 ± 0.24］ vs ［0.93 ± 0.10］ %, P<0.01) and Ⅱ (［6.72 ± 0.37］ vs ［2.95 ± 0.78］ %, P<0.01). After treatment, all the parameters were remarkably increased in groups C and D (grade a sperm: ［1.11 ± 0.30］ and ［0.60 ± 0.19］%; grade b sperm: ［2.40 ± 0.59］ and ［1.32 ± 0.27］ %; VSL: ［12.11 ± 1.62］ and ［11.47 ± 1.21］ μm/s; VCL: ［54.30 ± 2.35］ and ［45.75 ± 1.64］ μm/s; VAP ［18.40 ± 1.27］ and ［16.69 ± 1.02］ μm/s; expression of COXⅠ mRNA: ［1.86 ± 0.30］ and ［1.74 ± 0.17］ %) as compared with those in group B (P<0.05or P<0.01) except the COX activity and the expression of COX Ⅱ mRNA (P>0.05), and all the parameters were significantly higher in group C than in D (P<0.05or P<0.01). CONCLUSIONS UU infection can reduce grades a and b sperm, linear, curvilinear and mean sperm velocities, and the mRNA expressions of COX Ⅰ and Ⅱ while ZDD can improve these parameters. The improvement of sperm motility may not be associated with the activity of COX, and the COX activity may be related to the mRNA expression of COX II but not that of COXⅠ.
Animals ; Anti-Bacterial Agents ; therapeutic use ; Azithromycin ; therapeutic use ; Drugs, Chinese Herbal ; pharmacology ; Electron Transport Complex IV ; metabolism ; Epididymis ; drug effects ; enzymology ; Humans ; Male ; Mitochondria ; drug effects ; enzymology ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sperm Motility ; Spermatozoa ; drug effects ; enzymology ; physiology ; Ureaplasma Infections ; drug therapy ; enzymology ; Ureaplasma urealyticum

OBJECTIVETo explore the mechanisms of Qianjing Decoction in the treatment of oligoasthenospermia (OAS).

METHODSWe randomly divided 100 SPF male rats into five groups of equal number: normal, model, Huangjingzanyu, levocarnitine, and Qiangjing. OAS models were established in the animals followed by intragastrical administration of normal saline, ornidazole, Huangjingzanyu Capsules (200 mg per kg body weight per day), levocarnitine (100 mg per kg body weight per day), and Qianjing Decoction (10 g per kg body weight per day), respectively, qd, for 4 successive weeks. Then, we detected the concentration and motility of the epididymal sperm, obtained the contents of superoxide dismutase (SOD), malonaldehyde (MDA), glutathione peroxidase (GSH-Px), lactate dehydrogenase (LDH), α-glucosidase, and fructose in the epididymis, and determined the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and succinate dehydrogenase (SDH) in the epididymal tissue of the rats by real-time PCR.

RESULTSThe concentration and motility of the epididymal sperm in the model, Huangjingzanyu, levocarnitine, and Qianging groups were (35.34 ± 4.22) x 10(6)/ml and (40.04 ± 7.05)%, (48.12 ± 5.56) x 10(6)/ml and (62.46 ± 7.12)%, (47.14 ± 4.87) x 10(6)/ml and (63.23 ± 6.34)%, and (50.25 ± 5.08) x 10(6)/ml and (66.34 ± 7.58)%, respectively, all significantly lower than in the normal group ([53.05 ± 4.55] x 10(6)/ml and [70.20 ± 8.54]%) (P < 0.05), but remarkably higher in the Huangjingzanyu, levocarnitine, and Qiangjing groups than in the model rats (P < 0.05). Compared with the thinned epididymal lumen walls, decreased sperm count, and disorderly and loose arrangement of the lumens in the OAS models, the rats in the Huangjingzanyu, levocarnitine, and Qiangjing groups showed evidently thicker epididymal lumen walls, with the lumens full of sperm cells and arranged regularly and compactly, similar to those of the normal rats. The levels of SOD and GSH-Px were significantly lower but that of MDA markedly higher in the model rats ([84.12 ± 23.25], [10.56 ± 3.02], and [14.04 ± 2.06] nmol/mg) than in the normal group ([110.04 ± 19.56], [17.25 ± 3.56], and [8.87 ± 1.35] nmol/mg) (P < 0.05), while the former two indexes remarkably higher and the latter one significantly lower in the animals treated with Qiangjing Decoction ([120.56 ± 23.68], [16.34 ± 3.12], and [8.45 ± 1.56] nmol/mg), Huangjingzanyu Capsules ([115.34 ± 21.35], [15.23 ± 3.67], and [8.33 ± 1.54] nmol/mg), and levocarnitine ([116.67 ± 22.67], [15.35 ± 3.45], and [8.05 ± 1.78] nmol/mg) than in the models (P < 0.05). The levels of fructose, LDH and α-glucosidase were decreased markedly in the OAS models ([100.22 ± 12.12] mg/[ ml x g], [322 ± 46.13] U/[ ml x g], and [10.48 ± 2.33] U/[ml x g]) as compared with the normal rats ([128.12 ± 13.45] mg/[ml x g], [428 ± 35.12] U/[ml x g], and [15.34 ± 3.12] U/[ ml x g]) (P < 0.05), remarkably higher in the rats treated with Qiangjing ([130.23 ± 13.67] mg/[ml x g] [455 ± 51.50] U/[ml x g], and [18.56 ± 4.67] U/[ml x g]), Huangjingzanyu ([124.16 ± 14.02] mg/[ml x g], [ 419 ± 43.14] U/[ml x g], and [17.64 ± 4.08] U/[ml x g]), and levocarnitine ([123.34 ± 14.02] mg/[ml x g], [430 ± 31.80] U/ [ml x g], and [16.85 ± 5.55] U/[ml x g]) than in the models (P < 0.05). The Nrf2 mRNA expression was significantly reduced in the models as compared with the normal rats (P < 0.05) but remarkably increased in the Huangingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05). The SDH mRNA expression was significantly lower in the model than in the normal rats (P < 0.05) but markedly elevated in the Huangjingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05), remarkably higher in the Qiangjing than in the Huangjingzanyu group (P < 0.05).

CONCLUSIONOrnidazole induces OAS in rats, which is closely associated with excessive oxidation and energy metabolism dysfunction. Qiangjing Decoction can improve and even reverse ornidazole-induced OAS in rats as well as improve the ultrastructure of their testicular and epididymal tissues. Antioxidation and improvement of energy metabolism are probably the action mechanisms of Qiangjing Decoction in the treatment of OAS.

Animals ; Antioxidants ; Asthenozoospermia ; chemically induced ; drug therapy ; metabolism ; Carnitine ; pharmacology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Energy Metabolism ; drug effects ; Epididymis ; metabolism ; Glutathione Peroxidase ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Oligospermia ; chemically induced ; drug therapy ; metabolism ; Ornidazole ; Random Allocation ; Rats ; Sperm Count ; Sperm Motility ; Spermatozoa ; drug effects ; physiology ; Succinate Dehydrogenase ; metabolism ; Superoxide Dismutase ; metabolism ; alpha-Glucosidases ; metabolism

ObjectiveTo observe the clinical effect of Qilin Pills in the treatment of severe oligozoospermia after microsurgical ejaculatory duct reconstruction for obstructive azoospermia.

METHODSWe retrospectively analyzed 75 cases of obstructive azoospermia treated by ejaculatory duct reconstruction followed by administration of Qilin Pills. The patients were divided into a Qilin group (n=42) and a control group (n=33) postoperatively, treated with Qilin Pills and placebo, respectively. After 3 months of medication, we compared the sperm quality between the two groups of patients.

RESULTSAfter 3 months' treatment, all the patients experienced remarkable improvement in sperm quality (P<0.05). Compared with the controls, the patients in the Qilin group showed dramatic increases in sperm concentration, from (0.57±0.25) and (0.60±0.18) ×10⁶/ml before medication to (2.83±0.59) and (1.72 ±0.52) ×10⁶/ml after medication, significantly higher in the Qilin than in the control group (P<0.05). The percentage of grade a sperm was increased from (5.52±5.97) and (5.30±6.26)% to (11.56±9.96) and (10.27±6.52)%, that of grade a+b sperm from (9.68±8.63) and (8.64±10.10)% to (23.42 ±14.10) and (20.81±14.70)%, and that of morphologically normal sperm from (2.00±1.27) and (2.31±0.94)% to (3.54±2.47) and (3.47±1.33)%, respectively. No statistically significant differences were observed in sperm motility and normal sperm morphology between the two groups after treatment (P>0.05). The total effectiveness rate was higher in the Qilin group than in the controls (88.1% vs 72.7%), but with no significant difference between the two groups (P>0.05).

CONCLUSIONSQilin Pills are fairly effective in improving the quantity of sperm in obstructive azoospermia patients after ejaculatory duct reconstruction.

Adult ; Azoospermia ; drug therapy ; surgery ; Drugs, Chinese Herbal ; therapeutic use ; Ejaculatory Ducts ; surgery ; Humans ; Male ; Postoperative Complications ; drug therapy ; Retrospective Studies ; Sperm Count ; Sperm Motility ; Spermatozoa ; drug effects ; physiology

The detection of sperm DNA damage, as an important supplement to semen routine examination strategies, has been applied in some clinical andrology laboratories. What factors may lead to sperm DNA damage remains one of the concerns among many andrologists. Present studies show a variety of factors of sperm DNA damage, including age, environmental pollutants such as organophosphorus and organochloride pesticides, plasticizer, heavy metals such as lead, carcinogens such as polycyclic aromatic hydrocarbons (c-PAHs) and zearalenone (ZEA), male reproductive system diseases or systemic diseases such as varicocele, infection, tumor, spermatogenesis and maturation dysfunction, spinal cord injury and endocrine disorders, seasons and temperature, lifestyle, abstinence time, semen refrigeration, semen handling in vitro, and certain medications. Among them, spermatogenesis and sperm maturation dysfunction may be the most secretive factors, which are involved in the molecular mechanisms of sperm chromatin packaging and restructuring, such as the transformation of histone to protamine, single nucleotide polymorphism of genes, and the role of telomere, which may be one of the hotspots in the future studies of sperm DNA damage. Relevant researches in the future are expected to focus on the prevention of sperm DNA damage and clarification of its specific pathogenic mechanisms so as to provide some evidence for its treatment.
Age Factors ; Chromatin ; chemistry ; DNA Damage ; Environmental Pollutants ; toxicity ; Humans ; Male ; Protamines ; Semen ; drug effects ; Specimen Handling ; Spermatogenesis ; Spermatozoa ; drug effects ; Telomere ; physiology ; Varicocele ; complications

OBJECTIVETo investigate the protective effect of lycopene against cryopreservation injury of post-thawing human sperm and its mechanism.

METHODSSemen samples were collected from 25 volunteers, each sample equally divided into four parts to be cryopreserved with cryoprotectant only (Ly0 control) or cryoprotectant + lycopene at the concentrations of 2 (Ly2), 5 (Ly5), and 10 µmol/L (Ly10), respectively. Before and after thawing, the semen samples were subjected to computer-assisted semen analysis ( CASA) for sperm kinematics, flow cytometry for sperm apoptosis, thiobarbituric acid assay for malondialdehyde (MDA) concentration, and JC-1 fluorescent staining for the sperm mitochondrial membrane potential (MMP).

RESULTSAfter cryopreservation, sperm motility was markedly decreased in all the groups (P < 0.01). The rate of sperm apoptosis was significantly lower in the Ly5 group than in the Ly0 control ([25.68 ± 4.36]% vs [33.26 ± 4.78]%, P < 0.05), while sperm MMP remarkably higher in the former than in the latter ([66.18 ± 14.23]% vs [55.24 ± 12.31]%, P < 0.05). The Ly2, Ly5 and Ly10 groups showed no statistically significance differences in the MDA level from the Ly0 control (P > 0.05).

CONCLUSIONAddition of lycopene at a proper concentration to cryoprotectant may reduce oxidative damage to sperm mitochondria in the freezing-thawing process, attenuate oxidative stress injury induced by reactive oxygen species to sperm plasma membrane, and improve the anti-apoptosis ability of sperm.

Apoptosis ; Carotenoids ; pharmacology ; Cryopreservation ; Cryoprotective Agents ; pharmacology ; Flow Cytometry ; Humans ; Male ; Malondialdehyde ; analysis ; Oxidative Stress ; Reactive Oxygen Species ; Semen Analysis ; Semen Preservation ; adverse effects ; methods ; Sperm Motility ; Spermatozoa ; drug effects ; physiology