1.Regulatory effects of moxibustion at "Guanyuan" (CV4) on extragonadal estrogen and estrogen receptors in ovariectomized rats.
Qingchen ZHOU ; Xinyan GAO ; Kun LIU ; Bing ZHU
Chinese Acupuncture & Moxibustion 2025;45(12):1770-1776
OBJECTIVE:
To observe the regulatory effects of moxibustion at "Guanyuan" (CV4) on the synthesis of extragonadal estradiol (E2) and the expression of estrogen receptor (ER) in ovariectomized rats, aiming to explore the mechanism of moxibustion treatment for perimenopausal syndrome.
METHODS:
Forty-eight SD female rats of SPF grade were randomly divided into a sham-operation group, a model group and a moxibustion group, with 16 rats in each group. The model group and the moxibustion group underwent bilateral ovariectomy by the back incision method. Ten days after surgery, moxibustion was applied at "Guanyuan" (CV4) in the moxibustion group, 30 min each time, once a day for 10 days. After intervention, in the 3 groups, the body mass and uterus weight were measured; the serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH) and E2, as well as the skin and hypothalamus levels of E2 were detected by ELISA; the mRNA expression of aromatase (P450arom) in the skin and hypothalamus was detected by real-time PCR; the expression of ERα and ERβ in the hypothalamus, skin, and uterus was observed by immunofluorescence staining, and the density of positive cells was calculated using the Aipathwell digital pathology image analysis software.
RESULTS:
Compared with the sham-operation group, the body mass was increased (P<0.01) and the uterus weight was decreased (P<0.001) in the model group. Compared with the model group, the body mass was decreased in the moxibustion group (P<0.01). Compared with the sham-operation group, in the model group, the serum, hypothalamus and skin levels of E2 were decreased (P<0.01, P<0.05), while the serum levels of FSH and LH were increased (P<0.01); the expression of ERα and ERβ in the skin, hypothalamus and uterus was decreased (P<0.05, P<0.001). Compared with the model group, in the moxibustion group, the serum levels of E2 and LH, as well as the hypothalamus and skin levels of E2 were increased (P<0.05, P<0.01); the mRNA expression of P450arom, as well as the expression of ERα and ERβ in the skin and hypothalamus were increased (P<0.05).
CONCLUSION
Moxibustion at "Guanyuan" (CV4) reduces the body mass of ovariectomized rats by enhancing the synthesis of extragonadal E2 and increasing the expression of ER in the skin and hypothalamus, yet it does not alleviate uterine atrophy.
Animals
;
Female
;
Moxibustion
;
Rats
;
Ovariectomy
;
Acupuncture Points
;
Rats, Sprague-Dawley
;
Humans
;
Receptors, Estrogen/genetics*
;
Estrogens/metabolism*
;
Estradiol/metabolism*
;
Hypothalamus/metabolism*
;
Follicle Stimulating Hormone/blood*
;
Aromatase/genetics*
;
Luteinizing Hormone/blood*
;
Skin/metabolism*
2.Effect of "Zhibian" (BL54)-toward-"Shuidao" (ST28) acupuncture on reproductive function in mice with asthenozoospermia based on mitochondrial apoptosis.
Jianheng HAO ; Boya CHANG ; Jia REN ; Zhen GAO ; Yanlin ZHANG ; Haijun WANG ; Laixi JI
Chinese Acupuncture & Moxibustion 2025;45(1):71-81
OBJECTIVE:
To observe the effects of the "Zhibian" (BL54)-toward-"Shuidao" (ST28) acupuncture on key regulatory factors during mitochondrial apoptosis of testicular tissue in asthenozoospermia mice, and explore the potential mechanism of the protective effect of acupuncture on reproductive function.
METHODS:
Thirty C57BL/6 male mice were randomly divided into a blank group, a model group and an acupuncture group, 10 mice in each group. In the model and the acupuncture groups, the intraperitoneal injection of cyclophosphamide (30 mg•kg-1•d-1) was delivered for 7 days to prepare the asthenozoospermia model. After the success of modeling, the modeled mice in the acupuncture group were intervened with "Zhibian" (BL54)-toward-"Shuidao" (ST28) acupuncture, once daily and the needles were retained for 20 min. The duration of the intervention was 2 weeks. The general condition of each mouse was observed, and the body mass was recorded before modeling, after modeling and after intervention completion. After intervention, the testicular mass was recorded and the weight coefficient was calculated, and the mouse sperm quality was examined; the serum contents of testosterone (T), follicle stimulating hormone (FSH) and luteinizing hormone (LH) were detected using ELISA, the morphology of testicular tissue was observed using HE, the mitochondrial ultra-microstructure of testicular tissue was observed under transmission electrone microscopy, the mitochondrial membrane potential level of testicular tissue was detected using JC-1 staining, the positive rate of apoptosis cell of testicular tissue was observed using TUNEL; and the mRNA and protein expression of b-cell lymphocytoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), cytochrome c (Cyt C), apoptotic protease-activating factor1 (Apaf-1), Caspase-9 and Caspase-3 of testicular tissue was detected using real-time quantitative fluorescence PCR and Western blot methods separately; and the positive expression of Cleaved Caspase-3 of the testicular tissue was detected using immunohistochemistry.
RESULTS:
Compared with the blank group, the mice were in listless spirits, had shaggy hairs, the reduced appetite and movement, and weight loss in the model group (P<0.01); the testicular mass and the weight coefficient decreased (P<0.01); the total number of sperms, sperm motility, and sperm viability were declined (P<0.01); while the levels of serum T, FSH, and LH were dropped (P<0.01). The morphology of seminiferous tubules in testicular tissue was abnormal, the number of spermatogenic cells and the number of mitochondria decreased, the inner mitochondrial crest was fractured and lost, and vacuoles appeared. The level of mitochondrial membrane potential was reduced (P<0.01); and the positive rate of apoptosis cell in testicular tissue increased (P<0.01). The mRNA and protein expression of Bax, Cyt C, Apaf-1, Caspase-9 and Caspase-3 was elevated (P<0.01, P<0.05), the mRNA and protein expression of Bcl-2 was dropped (P<0.01), and the average absorbance value of Cleaved Caspase-3 increased (P<0.01). When compared with the model group, in the acupuncture group, the general condition of mice was improved, the testicular mass and the weight coefficient elevated (P<0.01); the total number of sperms, sperm motility, and sperm viability increased (P<0.01); while the levels of serum T, FSH, and LH rose (P<0.01). The pathological morphology of testicular tissue and the inner mitochondrial ultra-microstructure were ameliorated, the level of mitochondrial membrane potential was elevated (P<0.01); the positive rate of apoptosis cell was reduced (P<0.01). The mRNA and protein expression of Bax, Cyt C, Apaf-1, Caspase-9 and Caspase-3 was dropped (P<0.01, P<0.05), the mRNA and protein expression of Bcl-2 elevated (P<0.05), and the average absorbance value of Cleaved Caspase-3 declined (P<0.01).
CONCLUSION
"Zhibian" (BL54)-toward- "Shuidao" (ST28) acupuncture may ameliorate mouse reproductive function by inhibiting mitochondrial apoptosis pathway, alleviating testicular tissue damage in the asthenospermia mice induced by cyclophosphamide.
Animals
;
Male
;
Mice
;
Apoptosis
;
Acupuncture Therapy
;
Mitochondria/metabolism*
;
Asthenozoospermia/genetics*
;
Humans
;
Testis/metabolism*
;
Mice, Inbred C57BL
;
Spermatozoa/metabolism*
;
Acupuncture Points
;
Sperm Motility
;
Testosterone/blood*
;
Proto-Oncogene Proteins c-bcl-2/genetics*
;
Caspase 3/genetics*
;
Follicle Stimulating Hormone/blood*
;
Reproduction
;
Cytochromes c/genetics*
;
bcl-2-Associated X Protein/genetics*
;
Apoptotic Protease-Activating Factor 1/genetics*
3.Effects of acupuncture on the hypothalamic-pituitary-ovarian axis and FSH/cAMP signaling pathway in aged rats.
Yaoyao ZHU ; Yaqian YIN ; Huanfang XU ; Li YANG ; Weixin LI ; Chenchen SU ; Rong ZHANG ; Yigong FANG
Chinese Acupuncture & Moxibustion 2025;45(2):200-208
OBJECTIVE:
To explore the mechanism of acupuncture on improving ovarian hypofunction in aged rats from two perspectives: the overall regulation of the hypothalamic-pituitary-ovarian (HPO) axis and the local ovarian follicle stimulating hormone (FSH)/cyclic adenosine monophosphate (cAMP) signaling pathway.
METHODS:
Six 3-month-old female SPF-grade Sprague-Dawley (SD) rats were selected as the blank group. Another twelve 9-month-old female SD rats were randomly divided into a model group and an acupuncture group, with six rats in each. The acupuncture group received acupuncture at "Baihui" (GV20), "Guanyuan" (CV4), and bilateral "Ciliao" (BL32) for 20 min per session, once every other day, for a total of 10 sessions. Vaginal smear tests were performed daily to observe the estrous cycle of the rats. Ovarian morphology was observed using HE staining, and follicles at various stages were counted. ELISA was used to detect levels of serum FSH, luteinizing hormone (LH), estradiol (E2), anti-müllerian hormone (AMH), hypothalamic gonadotropin-releasing hormone (GnRH), pituitary FSH and LH, and ovarian cAMP. Immunohistochemistry and Western blot were used to detect the protein expression of ovarian cAMP protein kinase catalytic subunit, FSH receptor (FSHR), and P450. Real-time quantitative PCR was used to measure mRNA expression levels of FSHR and P450 in ovarian tissue.
RESULTS:
Compared with the blank group, the model group showed an increased rate of estrous cycle disorder (P<0.01), reduced granulosa cell layers with blurred boundaries and disordered arrangement, decreased numbers of developing follicles at all stages, and increased numbers of atretic follicles (P<0.01); the serum levels of FSH and LH were increased (P<0.01), while E2 and AMH levels were decreased (P<0.01); the hypothalamic GnRH and pituitary FSH and LH levels were elevated (P<0.01), and ovarian cAMP level was decreased (P<0.01); the positive expression and protein expression of ovarian P450, cAMP protein kinase catalytic subunit, and FSHR were reduced (P<0.01), and ovarian FSHR and P450 mRNA expression was decreased (P<0.01). Compared with the model group, the acupuncture group showed a reduced rate of estrous cycle disorder (P<0.01), clear granulosa cell margins, increased numbers of primordial and secondary follicles, and decreased numbers of atretic follicles (P<0.01); the serum FSH and LH levels were decreased (P<0.01, P<0.05), while E2 and AMH levels were increased (P<0.05, P<0.01); the hypothalamic GnRH and pituitary FSH and LH levels were decreased (P<0.01, P<0.05), and ovarian cAMP level was increased (P<0.01); the positive expression and protein expression of ovarian P450, cAMP protein kinase catalytic subunit, and FSHR were elevated (P<0.01), and ovarian FSHR and P450 mRNA expression was increased (P<0.01).
CONCLUSION
Acupuncture could delay ovarian hypofunction in aged rats, possibly through regulating the HPO axis and the FSH/cAMP signaling pathway.
Animals
;
Female
;
Rats
;
Rats, Sprague-Dawley
;
Follicle Stimulating Hormone/genetics*
;
Acupuncture Therapy
;
Ovary/metabolism*
;
Signal Transduction
;
Humans
;
Cyclic AMP/metabolism*
;
Hypothalamo-Hypophyseal System/metabolism*
;
Aging/metabolism*
;
Hypothalamus/metabolism*
;
Pituitary Gland/metabolism*
;
Gonadotropin-Releasing Hormone/metabolism*
4.Liuwei Dihuang Pills improve chemotherapy-induced ovarian injury in mice by promoting the proliferation of female germline stem cells.
Bo JIANG ; Wen-Yan ZHANG ; Guang-di LIN ; Xiao-Qing MA ; Guo-Xia LAN ; Jia-Wen ZHONG ; Ling QIN ; Jia-Li MAI ; Xiao-Rong LI
China Journal of Chinese Materia Medica 2025;50(9):2495-2504
This study primarily investigates the effect of Liuwei Dihuang Pills on the activation and proliferation of female germline stem cells(FGSCs) in the ovaries and cortex of mice with premature ovarian failure(POF), and how it improves ovarian function. ICR mice were randomly divided into the control group, model group, Liuwei Dihuang Pills group, Liuwei Dihuang Pills double-dose group, and estradiol valerate group. A mouse model of POF was established by intraperitoneal injection of cyclophosphamide. After successful modeling, the mice were treated with Liuwei Dihuang Pills or estradiol valerate for 28 days. Vaginal smears were prepared to observe the estrous cycle and body weight. After the last administration, mice were sacrificed and sampled. Serum levels of estradiol(E_2), follicle-stimulating hormone(FSH), luteinizing hormone(LH), and anti-Müllerian hormone(AMH) were measured by enzyme-linked immunosorbent assay(ELISA). Hematoxylin-eosin(HE) staining was used to observe ovarian morphology and to count follicles at all stages to evaluate ovarian function. Immunohistochemistry was used to detect the expression of mouse vasa homolog(MVH), a marker of ovarian FGSCs. Immunofluorescence staining, using co-labeling of MVH and proliferating cell nuclear antigen(PCNA), was used to detect the expression and localization of specific markers of FGSCs. Western blot was employed to assess the protein expression of MVH, octamer-binding transcription factor 4(Oct4), and PCNA in the ovaries. The results showed that compared with the control group, the model group exhibited disordered estrous cycles, decreased ovarian index, increased atretic follicles, and a reduced number of follicles at all stages. FSH and LH levels were significantly elevated, while AMH and E_2 levels were significantly reduced, indicating the success of the model. After treatment with Liuwei Dihuang Pills or estradiol valerate, hormone levels improved, the number of atretic follicles decreased, and the number of follicles at all stages increased. MVH marker protein and PCNA proliferative protein expression in ovarian tissue also increased. These results suggest that Liuwei Dihuang Pills regulate estrous cycles and hormone disorders in POF mice, promote the proliferation of FGSCs, improve follicular development in POF mice, and enhance ovarian function.
Animals
;
Female
;
Drugs, Chinese Herbal/administration & dosage*
;
Mice
;
Cell Proliferation/drug effects*
;
Mice, Inbred ICR
;
Ovary/cytology*
;
Primary Ovarian Insufficiency/genetics*
;
Follicle Stimulating Hormone/metabolism*
;
Humans
;
Anti-Mullerian Hormone/blood*
;
Antineoplastic Agents/adverse effects*
;
Luteinizing Hormone/metabolism*
;
Cyclophosphamide/adverse effects*
5.Incidence and genetic reproductive characteristics of AZFc microdeletion among patients with azoospermia or severe oligospermia.
Chiyan ZHOU ; Hui WANG ; Qin ZHU ; Luming WANG ; Binzhen ZHU ; Xiaodan LIU
Chinese Journal of Medical Genetics 2023;40(1):26-30
OBJECTIVE:
To explore the incidence of azoospermia factor c (AZFc) microdeletion among patients with azoospermia or severe oligospermia, its association with sex hormone/chromosomal karyotype, and its effect on the outcome of pregnancy following intracytoplasmic sperm injection (ICSI) treatment.
METHODS:
A total of 1 364 males with azoospermia or severe oligospermia who presented at the Affiliated Maternity and Child Health Care Hospital of Jiaxing College between 2013 and 2020 were subjected to AZF microdeletion and chromosome karyotyping analysis. The level of reproductive hormones in patients with AZFc deletions was compared with those of control groups A (with normal sperm indices) and B (azoospermia or severe oligospermia without AZFc microdeletion). The outcome of pregnancies for the AZFc-ICSI couples was compared with that of the control groups in regard to fertilization rate, superior embryo rate and clinical pregnancy rate.
RESULTS:
A total of 51 patients were found to harbor AZFc microdeletion, which yielded a detection rate of 3.74%. Seven patients also had chromosomal aberrations. Compared with control group A, patients with AZFc deletion had higher levels of PRL, FSH and LH (P < 0.05), whilst compared with control group B, only the PRL and FSH were increased (P < 0.05). Twenty two AZFc couples underwent ICSI treatment, and no significant difference was found in the rate of superior embryos and clinical pregnancy between the AZFc-ICSI couples and the control group (P > 0.05).
CONCLUSION
The incidence of AZFc microdeletion was 3.74% among patients with azoospermia or severe oligospermia. AZFc microdeletion was associated with chromosomal aberrations and increased levels of PRL, FSH and LH, but did not affect the clinical pregnancy rate after ICSI treatment.
Child
;
Humans
;
Male
;
Female
;
Pregnancy
;
Azoospermia/genetics*
;
Oligospermia/genetics*
;
Incidence
;
Chromosome Deletion
;
Chromosomes, Human, Y/genetics*
;
Semen
;
Infertility, Male/genetics*
;
Chromosome Aberrations
;
Follicle Stimulating Hormone/genetics*
6.Moxibustion alleviates decreased ovarian reserve in rats by restoring the PI3K/AKT signaling pathway.
Hong-Xiao LI ; Ling SHI ; Shang-Jie LIANG ; Chen-Chen FANG ; Qian-Qian XU ; Ge LU ; Qian WANG ; Jie CHENG ; Jie SHEN ; Mei-Hong SHEN
Journal of Integrative Medicine 2022;20(2):163-172
OBJECTIVE:
Moxibustion, a common therapy in traditional Chinese medicine, has potential benefits for treating decreased ovarian reserve (DOR). The present study investigates the protective effect of moxibustion in a rat model of DOR and explores the possible mechanisms.
METHODS:
Sixty-four female Sprague-Dawley rats were randomly divided into four groups: control, DOR, moxibustion (MOX), and hormone replacement therapy (HRT). The DOR rat model was established by intragastric administration of 50 mg/kg Tripterygium glycoside suspension (TGS), once daily for 14 days. MOX and HRT treatments were given from the day TGS administration was initiated. The ovarian reserve function was evaluated by monitoring the estrus cycle, morphological changes in ovaries, levels of serum estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and anti-Mullerian hormone (AMH), pregnancy rate and embryo numbers. Terminal-deoxynucleotidyl transferase-mediated nick-end-labeling staining was used to identify ovarian granulosa cell apoptosis, while the protein and mRNA expressions of Bax, B-cell lymphoma-2 (Bcl-2), phosphatidylinositol 3-kinase (PI3K) and protein kinase B (AKT) in ovarian tissues were examined by immunohistochemistry, Western blot and quantitative reverse transcription-polymerase chain reaction.
RESULTS:
Compared with the DOR group, MOX improved the disordered estrous cycle, promoted follicular growth, reduced the number of atresia follicles, increased the concentrations of serum E2 and AMH, and decreased serum FSH and LH concentrations. More importantly, the pregnancy rate and embryo numbers in DOR rats were both upregulated in the MOX treatment group, compared to the untreated DOR model. Further, we found that the MOX group had reduced apoptosis of ovarian granulosa cells, increased Bcl-2 expression and reduced expression of Bax. Furthermore, the PI3K/AKT signaling pathway was triggered by the moxibustion treatment.
CONCLUSION
Moxibustion improved ovarian function and suppressed apoptosis of ovarian granulosa cells in a rat model of DOR induced by TGS, and the mechanism may involve the PI3K/AKT signaling pathway.
Animals
;
Female
;
Follicle Stimulating Hormone
;
Luteinizing Hormone
;
Moxibustion
;
Ovarian Reserve
;
Phosphatidylinositol 3-Kinase/pharmacology*
;
Phosphatidylinositol 3-Kinases/metabolism*
;
Pregnancy
;
Proto-Oncogene Proteins c-akt/pharmacology*
;
Rats
;
Rats, Sprague-Dawley
;
Signal Transduction
;
bcl-2-Associated X Protein/genetics*
7.Altered microRNA profiles of testicular biopsies from patients with nonobstructive azoospermia.
Hai-Tao ZHANG ; Zhe ZHANG ; Kai HONG ; Wen-Hao TANG ; De-Feng LIU ; Jia-Ming MAO ; Yu-Zhuo YANG ; Hao-Cheng LIN ; Hui JIANG
Asian Journal of Andrology 2020;22(1):100-105
Many studies have shown that microRNAs (miRNAs) play vital roles during the spermatogenesis. However, little is known about the altered miRNA profiles of testicular tissues in nonobstructive azoospermia (NOA). Using microarray technology, the miRNA expression profiles of testicular biopsies from patients with NOA and of normal testicular tissues were determined. Bioinformatics analyses were conducted to predict the enriched biological processes and functions of identified miRNAs. The microarray data were validated by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), the results of which were then validated with a larger sample size. Correlations between the miRNA expression levels and clinical characteristics were analyzed. Receiver operating characteristic (ROC) curve analysis was used to evaluate the diagnostic ability of miRNAs for azoospermia. Hierarchical clustering showed that 129 miRNAs were significantly differentially expressed between the NOA and control groups. Bioinformatics analysis indicated that the differentially expressed miRNAs were involved in spermatogenesis, cell cycle, and mitotic prometaphase. In the subsequent qRT-PCR assays, the selected miRNA expression levels were consistent with the microarray results, and similar validated results were obtained with a larger sample size. Some clinical characteristics were significantly associated with the expression of certain miRNAs. In particular, we identified a combination of two miRNAs (miR-10b-3p and miR-34b-5p) that could serve as a predictive biomarker of azoospermia. This study provides altered miRNA profiles of testicular biopsies from NOA patients and examines the roles of miRNAs in spermatogenesis. These profiles may be useful for predicting and diagnosing the presence of testicular sperm in individuals with azoospermia.
Adult
;
Azoospermia/genetics*
;
Biopsy
;
Cluster Analysis
;
Computational Biology
;
Follicle Stimulating Hormone/metabolism*
;
Gene Expression Profiling
;
Humans
;
Luteinizing Hormone/metabolism*
;
Male
;
MicroRNAs/metabolism*
;
Reverse Transcriptase Polymerase Chain Reaction
;
Spermatogenesis/genetics*
;
Testis/metabolism*
;
Testosterone/metabolism*
;
Tissue Array Analysis
8.Identification of three novel SRD5A2 mutations in Chinese patients with 5α-reductase 2 deficiency.
Tong CHENG ; Hao WANG ; Bing HAN ; Hui ZHU ; Hai-Jun YAO ; Shuang-Xia ZHAO ; Wen-Jiao ZHU ; Hua-Ling ZHAI ; Fu-Guo CHEN ; Huai-Dong SONG ; Kai-Xiang CHENG ; Yang LIU ; Jie QIAO
Asian Journal of Andrology 2019;21(6):577-581
In this study, we investigated the genetics, clinical features, and therapeutic approach of 14 patients with 5α-reductase deficiency in China. Genotyping analysis was performed by direct sequencing of PCR products of the steroid 5α-reductase type 2 gene (SRD5A2). The 5α-reductase activities of three novel mutations were investigated by mutagenesis and an in vitro transfection assay. Most patients presented with a microphallus, variable degrees of hypospadias, and cryptorchidism. Eight of 14 patients (57.1%) were initially reared as females and changed their social gender from female to male after puberty. Nine mutations were identified in the 14 patients. p.G203S, p.Q6X, and p.R227Q were the most prevalent mutations. Three mutations (p.K35N, p.H162P, and p.Y136X) have not been reported previously. The nonsense mutation p.Y136X abolished enzymatic activity, whereas p.K35N and p.H162P retained partial enzymatic activity. Topical administration of dihydrotestosterone during infancy or early childhood combined with hypospadia repair surgery had good therapeutic results. In conclusion, we expand the mutation profile of SRD5A2 in the Chinese population. A rational clinical approach to this disorder requires early and accurate diagnosis, especially genetic diagnosis.
3-Oxo-5-alpha-Steroid 4-Dehydrogenase/genetics*
;
Adolescent
;
Adult
;
Asian People/genetics*
;
Child
;
Child, Preschool
;
China
;
Disorder of Sex Development, 46,XY/genetics*
;
Follicle Stimulating Hormone/blood*
;
Genitalia, Male/abnormalities*
;
Humans
;
Hypospadias/genetics*
;
Luteinizing Hormone/blood*
;
Male
;
Membrane Proteins/genetics*
;
Mutation/genetics*
;
Sequence Alignment
;
Steroid Metabolism, Inborn Errors/genetics*
;
Testosterone/blood*
;
Young Adult
9.Correlation of the single nucleotide polymorphisms rs34349826 and rs6521 of the LHB gene with male infertility in Chinese men.
Mao-Mao YU ; Li-Li GUO ; Pei-Ran ZHU ; Qiu-Yue WU ; Jing ZHANG ; Hong-Bo SUN ; Yu-Ling YIN ; Hui-Jie ZHI ; Li ZHOU ; Xin-Yi XIA
National Journal of Andrology 2018;24(6):509-515
ObjectiveTo study the relationship of the single nucleotide polymorphisms (SNP) rs34349826 (c.104 A>G) and rs6521 (c.114 C>G) of the luteinizing hormone beta-subunit (LHB) gene with male infertility in Chinese men.
METHODSThis case-control study included 405 males with primary infertility (the infertility group) and 424 normal fertile men (the control group), the former again divided into subgroups of oligospermia, severe oligozoospermia and azoospermia according to the sperm concentration. Clinical data were collected from all the subjects and genomic DNA obtained from their peripheral blood for genotyping rs34349826 and rs6521 of the LHB gene by Sequence MassArray. We analyzed the correlation of male infertility with the SNPs of the two loci using the logistic regression model as well as its association with their haplotype combination with the SHEsis online software.
RESULTSThere were statistically significant differences between the control and infertility groups in the semen volume ([3.51 ± 1.36] vs [3.74 ± 1.71] ml, P <0.05), sperm concentration ([79.21 ± 61.60] vs [27.37 ± 30.80] ×10⁶/ml, P <0.01), percentage of progressively motile sperm ([39.40 ± 9.64] % vs [11.90 ± 14.72] %, P <0.01), and levels of serum luteinizing hormone (LH) ([3.29 ± 1.39] vs [6.25 ± 4.83] IU/L, P <0.01) and follicle-stimulating hormone (FSH) ([4.56 ± 2.31] vs [15.64 ± 17.03] IU/L, P <0.01). Logistic regression analysis revealed no correlation between male infertility and the genotypes of the rs34349826 and rs6521 loci of the LHB gene, and similar results were found in the subgroups of the infertile males. SHEsis analysis on the haplotypes of the rs34349826 and rs6521 loci showed the GG genotype combination to be a protective factor against male infertility.
CONCLUSIONSThe rs34349826 and rs6521 loci of the LHB gene were not related to male infertility, which can be further confirmed by larger-sample studies. The GG genotype combination is a protective factor against male infertility.
Adult ; Azoospermia ; genetics ; Case-Control Studies ; China ; Follicle Stimulating Hormone ; Genotype ; Haplotypes ; Humans ; Infertility, Male ; genetics ; Logistic Models ; Luteinizing Hormone ; Luteinizing Hormone, beta Subunit ; genetics ; Male ; Oligospermia ; genetics ; Polymorphism, Single Nucleotide ; Sperm Count
10.Antagonistic effect of vitamin E on di-2-ethylhexyl phthalate-induced reproductive toxicity in male rats.
Chao-Yun WANG ; Juan-Juan ZHANG ; Peng DUAN
National Journal of Andrology 2018;24(7):589-595
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

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