Journal: International Journal of Nanomedicine

Article Title: Differential nanoreprotoxicity of silver nanoparticles in male somatic cells and spermatogonial stem cells

doi: 10.2147/IJN.S76062

Figure Lengend Snippet: Effect of AgNPs on the viability of TM3 and TM4 cells. Notes: TM3 and TM4 cells were treated with various concentrations of 10 nm ( A ) and 20 nm ( B ) AgNPs for 24 hours, and cytotoxicity was determined using the CCK-8 (top panel) and MTS (bottom panel) assays. The data are expressed as the mean ± SD of three independent experiments performed in triplicate; ** P <0.01. Abbreviations: AgNPs, silver nanoparticles; CCK, cell counting kit-8; IC 50 , half maximal inhibitory concentration; MTS, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; SD, standard deviation; TM3, Leydig; TM4, Sertoli.

Article Snippet: TM3 (KCLB No 21714) and TM4 (KCLB No 21715) cell lines were obtained from Korean cell line bank (Seoul, South Korea).

Techniques: CCK-8 Assay, Cell Counting, Concentration Assay, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Differential nanoreprotoxicity of silver nanoparticles in male somatic cells and spermatogonial stem cells

doi: 10.2147/IJN.S76062

Figure Lengend Snippet: Effect of AgNPs on LDH activity and ROS generation in TM3 and TM4 cells. Notes: TM3 and TM4 cells were treated with two different sizes of AgNPs: 10 nm ( A ) and 20 nm ( B ); LDH activity was assessed in the supernatants of cells treated or not with AgNPs and NAC using the LDH cytotoxicity detection kit. The y axis represents the absorbance as percentage of the controls, and the x axis shows IC 50 of AgNPs; ( C ) ROS generation in AgNPs (IC 50 )-treated TM3 and TM4 cells; DCF formation from H2DCF-DA was measured by fluorescence (excitation at 480 nm, emission at 530 nm). The data are expressed as the mean ± SD of three independent experiments performed in triplicate; ** P <0.01. Abbreviations: AgNPs, silver nanoparticles; DCF, 2′,7′-dichlorofluorescein; DMSO, dimethyl sulfoxide; H2DCF-DA, 2,7-dichlorodihydrofluorecein diacetate; H 2 O 2 , hydrogen peroxide; IC 50 , half maximal inhibitory concentration; LDH, lactate dehydrogenase; NAC, N-acetyl L-cysteine; ROS, reactive oxygen species; SD, standard deviation; TM3, Leydig; TM4, Sertoli.

Article Snippet: TM3 (KCLB No 21714) and TM4 (KCLB No 21715) cell lines were obtained from Korean cell line bank (Seoul, South Korea).

Techniques: Activity Assay, Fluorescence, Concentration Assay, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Differential nanoreprotoxicity of silver nanoparticles in male somatic cells and spermatogonial stem cells

doi: 10.2147/IJN.S76062

Figure Lengend Snippet: Cellular uptake of AgNPs induces accumulation of autophagosomes and autolysosomes. Notes: TM3 and TM4 cells were treated with AgNPs at IC 50 for 24 hours and analyzed by transmission electron microscopy (TEM). TEM images showed TM3 ( A – C , G – J ) and TM4 ( D , F , K – Q ) cells: not treated with AgNPs ( A , D ) and AgNP-treated showing crescent-like vacuoles ( B , E ), autophagic vacuoles ( C , F ), and accumulation of autophagosomes, autolysosomes, and damaged mitochondria ( G – J and K – Q ). Red arrows indicate the formation of autolysosomes. Autophagic vacuoles (AV) were classified into three types based on morphological criteria: AV-initial (AVi), intermediated (Avi/d), and degradative (Avd). All experiments were carried out in triplicate and repeated at least three times. Abbreviations: AgNPs, silver nanoparticles; Cy, cytoplasm; IC 50 , half maximal inhibitory concentration; N, nucleus; TM3, Leydig; TM4, Sertoli; TAV, typical autophagic vacuoles; DM, damaged mitochondria; MVB, multivesicular bodies.

Article Snippet: TM3 (KCLB No 21714) and TM4 (KCLB No 21715) cell lines were obtained from Korean cell line bank (Seoul, South Korea).

Techniques: Transmission Assay, Electron Microscopy, Concentration Assay

Journal: International Journal of Nanomedicine

Article Title: Differential nanoreprotoxicity of silver nanoparticles in male somatic cells and spermatogonial stem cells

doi: 10.2147/IJN.S76062

Figure Lengend Snippet: AgNPs accelerate expression of autophagy genes. Notes: TM3 ( A ) and TM4 ( B ) cells were treated with AgNPs for 24 hours and analyzed for the expression of autophagy-related genes; for each treatment condition and time point, mRNA expression of AgNP-treated cells was normalized to that of untreated cells. The data are expressed as the mean relative gene expression ± SD of three independent experiments performed in triplicate; ** P <0.01. Abbreviations: AgNPs, silver nanoparticles; mRNA, messenger RNA; SD, standard deviation; TM3, Leydig; TM4, Sertoli.

Article Snippet: TM3 (KCLB No 21714) and TM4 (KCLB No 21715) cell lines were obtained from Korean cell line bank (Seoul, South Korea).

Techniques: Expressing, Gene Expression, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Differential nanoreprotoxicity of silver nanoparticles in male somatic cells and spermatogonial stem cells

doi: 10.2147/IJN.S76062

Figure Lengend Snippet: AgNPs promote ROS-induced apoptosis. Notes: TM3 and TM4 cells were pretreated with 10 mM NAC for 1 hour and then treated with AgNPs for 6 hours ( A , B ); TM3 ( A ) and TM4 ( B ) cells were stained with Annexin V (x axis) and PI (y axis) and analyzed for cell death by flow cytometry using a FACSCanto TMII; the percentages of apoptotic cells are shown in each quadrant ( C , D ); TM3 ( C ) and TM4 ( D ) cells were exposed to the respective IC 50 of AgNPs for 6 hours and analyzed for protein expression by western blotting using the indicated antibodies; ( E , F ) protein expression relative to β-actin used as the loading control; ( G , H ) relative mRNA expression levels of apoptotic genes analyzed by RT-PCR. The data are expressed as the mean relative gene expression ± SD of three independent experiments performed in triplicate; * P <0.05, ** P <0.01. Abbreviations: AgNPs, silver nanoparticles; IC 50 , half maximal inhibitory concentration; mRNA, messenger RNA; NAC, N-acetyl L-cysteine; PI, propidium iodide; ROS, reactive oxygen species; RT-PCR, real-time polymerase chain reaction; SD, standard deviation; TM3, Leydig; TM4, Sertoli.

Article Snippet: TM3 (KCLB No 21714) and TM4 (KCLB No 21715) cell lines were obtained from Korean cell line bank (Seoul, South Korea).

Techniques: Staining, Flow Cytometry, Expressing, Western Blot, Control, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Concentration Assay, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Differential nanoreprotoxicity of silver nanoparticles in male somatic cells and spermatogonial stem cells

doi: 10.2147/IJN.S76062

Figure Lengend Snippet: Effect of AgNPs on gene expression in TM3 and TM4 cells. Notes: ( A ) Quantitative (q) RT-PCR for testosterone biosynthesis-related genes in TM3 cells; ( B ) qRT-PCR for the BTB genes Gdnf , Tgfβ1 , Ar , and Amh in TM4 cells. The expression of all genes was normalized to that of Gapdh . The data are expressed as the mean relative gene expression ± SD of three independent experiments performed in triplicate; * P <0.05, ** P <0.01. Abbreviations: AgNPs, silver nanoparticles; BTB, blood–testes barrier; mRNA, messenger RNA; RT-PCR, real-time polymerase chain reaction; SD, standard deviation; TM3, Leydig; TM4, Sertoli; qRT-PCR, quantitative RT-PCR.

Article Snippet: TM3 (KCLB No 21714) and TM4 (KCLB No 21715) cell lines were obtained from Korean cell line bank (Seoul, South Korea).

Techniques: Gene Expression, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Differential nanoreprotoxicity of silver nanoparticles in male somatic cells and spermatogonial stem cells

doi: 10.2147/IJN.S76062

Figure Lengend Snippet: Effects of AgNPs on the proliferation and differentiation of SSCs. Notes: ( A ) FACS analysis of SSCs using CD49f antibody; ( B ) the percentage of SSCs cultured on TM3 or TM4 as feeder cells; ( C ) expression of the genes involved in meiosis, self-renewal, and differentiation assessed by quantitative RT-PCR and normalized to that of Gapdh. Incubation of SSCs on TM3 and TM4 cells treated with AgNP (10 μg/mL) caused a significant decrease in mRNA levels of the analyzed genes. The results are presented as mean ± SEM of triplicate measurements; ** P <0.01. Abbreviations: AgNPs, silver nanoparticles; FACS, fluorescence-activated cell sorting; RT-PCR, real-time polymerase chain reaction; SEM, standard error of the mean; SSCs, spermatogonial stem cells; TM3, Leydig; TM4, Sertoli; FSC-H, forward scatter-height.

Article Snippet: TM3 (KCLB No 21714) and TM4 (KCLB No 21715) cell lines were obtained from Korean cell line bank (Seoul, South Korea).

Techniques: Cell Culture, Expressing, Quantitative RT-PCR, Incubation, Fluorescence, FACS, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Journal: Oxidative Medicine and Cellular Longevity

Article Title: A Protein from Dioscorea polystachya (Chinese Yam) Improves Hydrocortisone-Induced Testicular Dysfunction by Alleviating Leydig Cell Injury via Upregulation of the Nrf2 Pathway

doi: 10.1155/2021/3575016

Figure Lengend Snippet: Amelioration of testicular dysfunction by DP1 was related to the restoration of Leydig cells. (a) Immunohistochemistry with an antibody against 3 β -HSD was performed to measure the amount of Leydig cells in testicular tissues of rats. (b) Effect of DP1 on the cell viability of TM3 cells detected by CCK-8 assays. (c, d) Flow cytometry was conducted to analyze the number of apoptotic TM3 cells. (e, g) The ratio of Bcl-2/Bax in TM3 cells was detected by Western blotting. (f, g) The protein expression levels of TGF- β 1 and p-SMAD2/3 in TM3 cells were detected by Western blotting. (h) Immunofluorescence assays were performed to detect the expression of TGF- β 1 in TM3 cells. (i) Comparison of the testosterone levels in TM3 cells. Each bar represents the mean ± SEM. All images were captured at a magnification of ×100. # P < 0.05, ## P < 0.01, and ### P < 0.001 vs. the control group; ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 vs. the H 2 O 2 group.

Article Snippet: TM3 cells were cultured in TM3 cell culture medium (Procell, Wuhan, China) at 37°C.

Techniques: Immunohistochemistry, CCK-8 Assay, Flow Cytometry, Western Blot, Expressing, Immunofluorescence, Comparison, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: A Protein from Dioscorea polystachya (Chinese Yam) Improves Hydrocortisone-Induced Testicular Dysfunction by Alleviating Leydig Cell Injury via Upregulation of the Nrf2 Pathway

doi: 10.1155/2021/3575016

Figure Lengend Snippet: Inhibition of testicular dysfunction by DP1 was dependent on the reduction in oxidative stress in Leydig cells. (a) 8-OHdG levels and (b) SOD activities in testicular tissues. (c) Immunofluorescence staining was performed to measure the superoxide anion levels in TM3 cells. (d, e) The ratio of Bcl-2/Bax and the protein expression levels of TGF- β 1 and p-SMAD2/3 in TM3 cells were detected by Western blotting. (f) Comparison of the testosterone levels in TM3 cells. Each bar represents the mean ± SEM. The images were captured at a magnification of ×100. # P < 0.05, ## P < 0.01, and ### P < 0.001 vs. the control group; ∗ P < 0.05 and ∗∗∗ P < 0.001 vs. the HCT group or H 2 O 2 group; ^ P < 0.05 and ^^^ P < 0.001 vs. the DP1 group. XO: xanthine oxidase.

Article Snippet: TM3 cells were cultured in TM3 cell culture medium (Procell, Wuhan, China) at 37°C.

Techniques: Inhibition, Immunofluorescence, Staining, Expressing, Western Blot, Comparison, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: A Protein from Dioscorea polystachya (Chinese Yam) Improves Hydrocortisone-Induced Testicular Dysfunction by Alleviating Leydig Cell Injury via Upregulation of the Nrf2 Pathway

doi: 10.1155/2021/3575016

Figure Lengend Snippet: DP1 promoted the upregulation of the Nrf2/HO-1 pathway. (a, b) The protein expression levels of Nrf2 and HO-1 in testicular tissues were detected by Western blotting. (c) Relative mRNA expression of HO-1, NQO1, and GCLC normalized to GAPDH in TM3 cells. (d, e) The protein expression levels of total Nrf2, nuclear Nrf2, and nuclear HO-1 in TM3 cells were detected by Western blotting. Each bar represents the mean ± SEM. The images were captured at a magnification of ×100. # P < 0.05 and ### P < 0.001 vs. the control group; ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 vs. the HCT group or H 2 O 2 group.

Article Snippet: TM3 cells were cultured in TM3 cell culture medium (Procell, Wuhan, China) at 37°C.

Techniques: Expressing, Western Blot, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: A Protein from Dioscorea polystachya (Chinese Yam) Improves Hydrocortisone-Induced Testicular Dysfunction by Alleviating Leydig Cell Injury via Upregulation of the Nrf2 Pathway

doi: 10.1155/2021/3575016

Figure Lengend Snippet: Nrf2 activation was required for DP1 to inhibit the Leydig cell injury. (a) Real-time PCR analysis of Nrf2 mRNA levels in TM3 cells transfected with Nrf2 siRNA or NC siRNA for 48 h. (b) Immunofluorescence staining was performed to measure the superoxide anion levels in TM3 cells. (c, d) The protein expression levels of Nrf2 and HO-1 in testicular tissues were detected by Western blotting. (e, f) The ratio of Bcl-2/Bax and the protein expression levels of TGF- β 1 and p-SMAD2/3 in TM3 cells were detected by Western blotting. (g) Immunofluorescence analysis of TGF- β 1 expression in TM3 cells. (h) Comparison of the testosterone levels in TM3 cells. Each bar represents the mean ± SEM. All images were captured at a magnification of ×100. ## P < 0.01 and ### P < 0.001 vs. the control group; ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 vs. the DP1 group.

Article Snippet: TM3 cells were cultured in TM3 cell culture medium (Procell, Wuhan, China) at 37°C.

Techniques: Activation Assay, Real-time Polymerase Chain Reaction, Transfection, Immunofluorescence, Staining, Expressing, Western Blot, Comparison, Control

Journal: International Journal of Environmental Research and Public Health

Article Title: Effects and Mechanisms of Phthalates’ Action on Reproductive Processes and Reproductive Health: A Literature Review

doi: 10.3390/ijerph17186811

Figure Lengend Snippet: The function of some genes regulating male and female reproductive system development [ 242 ].

Article Snippet: In vitro studies demonstrated that DEHP exposure at 40, 80 and 160 μM and dibutyl phthalate (DBP) exposure at 10 and 100 mg/L caused the apoptosis of TM3 Leydig cells and Sertoli cells of Male Sprague-Dawley rats, respectively [ , ].

Techniques: Expressing, Inhibition

Journal: Molecules

Article Title: Vegetables and Their Bioactive Compounds as Anti-Aging Drugs

doi: 10.3390/molecules27072316

Figure Lengend Snippet: Anti-aging properties of vegetables.

Article Snippet: Taraxacum officinale F.H. Wigg. (Dandelion) , Aqueous extract , TM3 cells, an immature mouse Leydig cell line, and 18-week-old male Sprague Dawley rats , Western blot analysis Measurement of serum testosterone level Swimming retention test Measurement of sperm count and activity , Protected TM3 cells from serum restriction and oxidative stress via activation of ERK and Akt pathways Improved testosterone level and activation of spermatogenesis in rats Improved physical locomotion Improved quality of life for aging males , [ ] .

Techniques: FRAP Assay, Activity Assay, Clinical Proteomics, In Vitro, DPPH Assay, Concentration Assay, Western Blot, Expressing, Antioxidant Activity Assay, Cell Culture, Irradiation, Quantitative RT-PCR, Inhibition, Migration, Spectroscopy, Isolation, Gene Expression, Histopathology, Cell Counting, Ointment, High Performance Liquid Chromatography, Solubility, Activation Assay, RNA Extraction, Enzyme-linked Immunosorbent Assay, Cytotoxicity Assay, Synthesized, Sequencing, Immunohistochemical staining, In Vivo, Formulation, Pore Size, Spot Test, Transformation Assay, ABTS Assay, ROS Assay, MTT Assay, Viability Assay, Enzymatic Assay