Journal: Cell Communication and Signaling : CCS

Article Title: Dual deficiency of melatonin and dihydrotestosterone promotes stromal cell damage and mediates prostatitis via the cGAS-STING pathway in sleep-deprived mice

doi: 10.1186/s12964-024-01554-5

Figure Lengend Snippet: Characteristics and proteomic analysis of prostate in mice after 3 weeks of SD. A Photos of prostate and corresponding sizes (prostate index) analysis. B HE staining showing the typical pathological changes of prostatitis. C Quantification of urinary frequency by urine spots assay. D-G ELISA analysis of cytokines in serum, including IL-6, TNF-α, PGE2 and IFN-β. H-K ELISA analysis of cytokines in prostate tissue, including IL-6, TNF-α, PGE2 and IFN-β. Proteomic analysis of control and SD mice including ( L ) KEGG pathway enrichment, ( M ) hierarchical clustering heatmap, and ( N ) volcano plot of differentially expressed proteins. O Proteomic analysis showing the underlying mechanism of prostatitis induction. P Analysis of MT level in serum after 3 weeks of SD. Q-R Analysis of DHT level in serum and prostate tissue after 3 weeks of SD. S Schematic hypothesis of the cGAS-STING pathway activation by the dual deficiency of MT and DHT for inducing inflammatory microenvironment in prostate. Data were presented as means ± SEM ( n = 5). Statistical significance was calculated using the Student’s t test ( A , C , D - K , and P - R ). ns, no significance, ** P < 0.01, and *** P < 0.001

Article Snippet: Cytokine levels in mouse plasma and prostate tissue homogenate samples were detected using ELISA kits for MT (E-EL-M0788c, Elabscience, Wuhan, China), DHT (E-EL-0031c, Elabscience, Wuhan, China), E2 (E-OSEL-M0008, Elabscience, Wuhan, China), T (E-OSEL-M0003, Elabscience, Wuhan, China), GnRH (E-EL-0071c, Elabscience, Wuhan, China), LH (E-EL-M3053, Elabscience, Wuhan, China), IFN-β (EK2236-96, MULTISCIENCES, Hangzhou, China), TNF-ɑ (EK282/4–96, MULTISCIENCES, Hangzhou, China), IL-6 (EK206/3–96, MULTISCIENCES, Hangzhou, China), and PGE2 (EK8103/2–96, MULTISCIENCES, Hangzhou, China).

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Control, Activation Assay

Journal: Cell Communication and Signaling : CCS

Article Title: Dual deficiency of melatonin and dihydrotestosterone promotes stromal cell damage and mediates prostatitis via the cGAS-STING pathway in sleep-deprived mice

doi: 10.1186/s12964-024-01554-5

Figure Lengend Snippet: Induction of cGAS-STING pathway-mediated prostatitis by co-inhibiting MT and DHT in normal mice. A Schematic illustration of treatment by MT inhibitor (4-P-PDOT) and DHT inhibitor (finasteride) in normal mice for 3 weeks. 4-P-PDOT and finasteride were used to establish the mice model with dual deficient MT and DHT, which was applied to verify the relationship between dual deficient MT and DHT and prostatitis in SD mice. B-C Photos of prostate and corresponding sizes (prostate index) analysis. D The level of DHT in prostate of mice treated by finasteride. E Changes of tactile allodynia by Von Frey filaments. F HE staining showing the changes of prostate in mice after 4-P-PDOT and finasteride treatment. Scale bar, 100 μm. G-H The indexes of oxidative stress in prostate, including MDA and SOD. I-K ELISA analysis of cytokines in serum, including IL-6, TNF-α, and PGE2. L-N ELISA analysis of cytokines in prostate tissue, including IL-6, TNF-α, and PGE2. O-P Immunohistochemical staining of p-TBK1 and p-IRF3. Scale bar, 100 μm. Q-R The corresponding average optical density (AOD) of immunohistochemistry for p-TBK1 and p-IRF3. S-T The changes of IFN-β levels in serum and prostate tissue. Data were presented as means ± SEM ( n = 5). Statistical significance was calculated using the one-way ANOVA ( C-E , G-N , and Q-T ). ns, no significance, and *** P < 0.001

Article Snippet: Cytokine levels in mouse plasma and prostate tissue homogenate samples were detected using ELISA kits for MT (E-EL-M0788c, Elabscience, Wuhan, China), DHT (E-EL-0031c, Elabscience, Wuhan, China), E2 (E-OSEL-M0008, Elabscience, Wuhan, China), T (E-OSEL-M0003, Elabscience, Wuhan, China), GnRH (E-EL-0071c, Elabscience, Wuhan, China), LH (E-EL-M3053, Elabscience, Wuhan, China), IFN-β (EK2236-96, MULTISCIENCES, Hangzhou, China), TNF-ɑ (EK282/4–96, MULTISCIENCES, Hangzhou, China), IL-6 (EK206/3–96, MULTISCIENCES, Hangzhou, China), and PGE2 (EK8103/2–96, MULTISCIENCES, Hangzhou, China).

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Immunohistochemistry

Journal: Cell Communication and Signaling : CCS

Article Title: Dual deficiency of melatonin and dihydrotestosterone promotes stromal cell damage and mediates prostatitis via the cGAS-STING pathway in sleep-deprived mice

doi: 10.1186/s12964-024-01554-5

Figure Lengend Snippet: Progression of prostatitis in sleep-deprived mice after sleep recovery. A Schematic illustration of sleep recovery for different time (1 to 3 weeks) in SD mice. B Analysis of MT level in serum in SD mice after 3 weeks of sleep-recovery (R1, R2, and R3). C Analysis of DHT level in prostate tissue after 3 weeks of sleep-recovery. D The prostate sizes (prostate index) analysis. E-G ELISA analysis of cytokines in prostate, including IL-6, TNF-α, and PGE2. H-I The concentration of IFN-β in serum and prostate tissue. J HE staining showing the changes of prostate in SD mice after 1 week of sleep-recovery. Scale bar, 100 μm. K The corresponding inflammation score analysis of prostate in HE staining. L Evaluation of tactile allodynia by Von Frey filaments in mice. M-N Western blot and corresponding analysis of the essential proteins of cGAS-STING pathway in prostate of mice after 1 week of sleep-recovery. O Schematic illustration of sleep re-deprivation for different time (1, 3 and 7 days) in mice. P HE staining revealing the changes of prostate in sleep-recovery mice after sleep re-deprivation (R-SD) for different periods (1, 3, and 7 days). Scale bar, 100 μm. Data were presented as means ± SEM ( n ≥ 5). Statistical significance was calculated using the one-way ANOVA ( B - I , K , L , and N ). ns, no significance, * P < 0.05, ** P < 0.01, and *** P < 0.001

Article Snippet: Cytokine levels in mouse plasma and prostate tissue homogenate samples were detected using ELISA kits for MT (E-EL-M0788c, Elabscience, Wuhan, China), DHT (E-EL-0031c, Elabscience, Wuhan, China), E2 (E-OSEL-M0008, Elabscience, Wuhan, China), T (E-OSEL-M0003, Elabscience, Wuhan, China), GnRH (E-EL-0071c, Elabscience, Wuhan, China), LH (E-EL-M3053, Elabscience, Wuhan, China), IFN-β (EK2236-96, MULTISCIENCES, Hangzhou, China), TNF-ɑ (EK282/4–96, MULTISCIENCES, Hangzhou, China), IL-6 (EK206/3–96, MULTISCIENCES, Hangzhou, China), and PGE2 (EK8103/2–96, MULTISCIENCES, Hangzhou, China).

Techniques: Enzyme-linked Immunosorbent Assay, Concentration Assay, Staining, Western Blot

Journal: International journal of molecular medicine

Article Title: Development of finasteride polymer microspheres for systemic application in androgenic alopecia.

doi: 10.3892/ijmm.2019.4149

Figure Lengend Snippet: Figure 2. FNS‑loaded microspheres effectively prevent 5α‑reductase activity in a testosterone‑induced androgenic alopecia mouse model. (A) Schematic diagram of the study protocol. The mice were administered with the test compounds orally or subcutaneously for 8 weeks. (B) Images of the dorsal skin at 8 weeks. Individual data were obtained using enzyme‑linked immunosorbent assays for (C) testosterone and (D) DHT in serum. The DHT levels decreased 4 weeks after orally‑applied FNS and FNS‑loaded microspheres administration. All data are presented as the mean ± standard error of the mean from three independent experiments. *P<0.05 and **P<0.01, vs. microspheres‑treated group. FNS, finasteride; TP, testosterone propionate; DHT, dihydrotestosterone.

Article Snippet: Serum testosterone (cat. no. CSB‐E05101m) and DHT (cat. no. CSB‐E07880m) were measured via enzyme linked immunosorbent assays (ELISA; Cusabio, Wuhan, China).

Techniques: Activity Assay

Journal: Foods (Basel, Switzerland)

Article Title: Analysis of the Beneficial Effects of Probiotics on the Gut-Prostate Axis Using Prostatic Co-Culture Model.

doi: 10.3390/foods13223647

Figure Lengend Snippet: Figure 2. Permeability and adhesion study on CaCo-2 cells. In (A), Cell viability is measured by using the MTT test; in (B), TEER Value using EVOM3; in (C), SCFA analysis is conducted using an ELISA kit; in (D), butyric acid analysis is carried out using an ELISA kit; in (E) the analysis of aggregation activity and in (F) the analysis of hydrophobicity activity. Data are mean ± SD of five independent experiments performed in triplicates compared to control values (0% line). In (A,C–F), * p < 0.05 vs. control; ** p < 0.05 vs. single agents. In (B), p < 0.05 vs. control; ** p < 0.05 vs. single agents.

Article Snippet: According to the manufacturer, a Human Dihydrotestosterone ELISA Kit (Cusabio, Houston, TX, USA) was used to measure DHT in co-culture prostatic supernatants [43].

Techniques: Permeability, Enzyme-linked Immunosorbent Assay, Activity Assay, Control

Journal: Foods (Basel, Switzerland)

Article Title: Analysis of the Beneficial Effects of Probiotics on the Gut-Prostate Axis Using Prostatic Co-Culture Model.

doi: 10.3390/foods13223647

Figure Lengend Snippet: Figure 3. Integrity analysis. In (A), the analysis of claudin-1 measured by ELISA test; in (B), the analysis of ZO-1 by ELISA test; in (C), the analysis of occludin-1 by ELISA kit. Data are mean ± SD of five independent experiments performed in triplicates vs. control values (0% line) and * p < 0.05 vs. control; ** p < 0.05 vs. single agents.

Article Snippet: According to the manufacturer, a Human Dihydrotestosterone ELISA Kit (Cusabio, Houston, TX, USA) was used to measure DHT in co-culture prostatic supernatants [43].

Techniques: Enzyme-linked Immunosorbent Assay, Control

Journal: Foods (Basel, Switzerland)

Article Title: Analysis of the Beneficial Effects of Probiotics on the Gut-Prostate Axis Using Prostatic Co-Culture Model.

doi: 10.3390/foods13223647

Figure Lengend Snippet: Figure 4. Analysis of oxidative stress and inflammation. All of the following results are derived from the ELISA kit: (A) ROS generation; (B) SOD levels; (C) TNF-α production; (D) IL-6 production; and (E) IL-10 production. The mean ± SD of five separate studies conducted in triplicate is shown, along with * p < 0.05 vs. control, ** p < 0.05 vs. single drugs, and # p < 0.05 vs. DHT, in comparison to control values (0% line).

Article Snippet: According to the manufacturer, a Human Dihydrotestosterone ELISA Kit (Cusabio, Houston, TX, USA) was used to measure DHT in co-culture prostatic supernatants [43].

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Control, Comparison

Journal: Foods (Basel, Switzerland)

Article Title: Analysis of the Beneficial Effects of Probiotics on the Gut-Prostate Axis Using Prostatic Co-Culture Model.

doi: 10.3390/foods13223647

Figure Lengend Snippet: Figure 5. Mechanism analyses for prostate hyperplasia. Testosterone levels (A), 5α-reductase levels (B), DHT levels (C), AR levels (D), and PSA levels (E) are all derived from the ELISA kit. Values are mean ± SD from five independent tests in triplicates compared to control values (0% line). * p < 0.05 vs. control; ** p < 0.05 vs. single agents; # p < 0.05 vs. DHT.

Article Snippet: According to the manufacturer, a Human Dihydrotestosterone ELISA Kit (Cusabio, Houston, TX, USA) was used to measure DHT in co-culture prostatic supernatants [43].

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: Foods (Basel, Switzerland)

Article Title: Analysis of the Beneficial Effects of Probiotics on the Gut-Prostate Axis Using Prostatic Co-Culture Model.

doi: 10.3390/foods13223647

Figure Lengend Snippet: Figure 6. Serotonin and proliferation studies. The following results are derived from an ELISA kit: (A) 5-Htr1a levels, (B) serotonin production, (C) Ki67 expression, and (D) proliferation rate. The mean ± SD of five separate, duplicate tests is presented, along with * p < 0.05 vs. control, ** p < 0.05 vs. single drugs, and # p < 0.05 vs. DHT, in comparison to control values (0% line).

Article Snippet: According to the manufacturer, a Human Dihydrotestosterone ELISA Kit (Cusabio, Houston, TX, USA) was used to measure DHT in co-culture prostatic supernatants [43].

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Expressing, Control, Comparison

Journal: International Journal of Endocrinology and Metabolism

Article Title: Accuracy of Urinary Etiocholanolone/Androsterone Ratio as Alternative to Serum Testosterone/Dihydrotestosterone Ratio for Diagnosis of 5 Alpha-reductase Type 2 Deficiency Patients and Carriers in Indonesia

doi: 10.5812/ijem.109510

Figure Lengend Snippet: Comparison of ROC Curve of T/DHT and Et/An ratios in diagnosing cases (A) and Carriers (B). E/A, urinary etiocholanolone/androsterone ratio; ROC, receiver operating curve; T/DHT, testosterone/dihydrotestosterone ratio.

Article Snippet: We performed the competitive ELISA methods as described in our previous report ( ) using the TOSOH AIA-900 automated immunoassay analyzer to examine the serum testosterone (ST AIA-PACK testosterone) and commercial kit of DH-optimized ELISA for DHT test (DRG Instruments GmbH, Germany, ref. EIA-5761).

Techniques:

Journal: International Journal of Endocrinology and Metabolism

Article Title: Accuracy of Urinary Etiocholanolone/Androsterone Ratio as Alternative to Serum Testosterone/Dihydrotestosterone Ratio for Diagnosis of 5 Alpha-reductase Type 2 Deficiency Patients and Carriers in Indonesia

doi: 10.5812/ijem.109510

Figure Lengend Snippet: T/DHT ratio in prepubertal and pubertal cases. 5ARD2, 5 alpha-reductase deficiency type 2; T, testosterone; DHT, dihydrotestosterone

Article Snippet: We performed the competitive ELISA methods as described in our previous report ( ) using the TOSOH AIA-900 automated immunoassay analyzer to examine the serum testosterone (ST AIA-PACK testosterone) and commercial kit of DH-optimized ELISA for DHT test (DRG Instruments GmbH, Germany, ref. EIA-5761).

Techniques:

Journal: Nutrition Research and Practice

Article Title: Corni Fructus attenuates testosterone-induced benign prostatic hyperplasia by suppressing 5α-reductase and androgen receptor expression in rats

doi: 10.4162/nrp.2018.12.5.378

Figure Lengend Snippet: The serum concentrations of testosterone (A) and DHT (B) were examined by ELISA. The data shown represent the mean ± SEM of six rats per group ( P < 0.05). BPH, benign prostatic hyperplasia; CF, Corni Fructus water extract; FINA, finasteride; DHT, dihydrotestosterone.

Article Snippet: Serum concentrations of testosterone and DHT were determined by using Testosterone enzyme-linked immunosorbent assay (ELISA) kit (BioVision, Inc., Milpitas, CA, USA) and rat DHT ELISA kit (Mybiosource Inc., San Diego, CA, USA), according to the manufacturer's instructions, which are based on the principle of competitive inhibition enzyme immunoassay.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Nutrition Research and Practice

Article Title: Corni Fructus attenuates testosterone-induced benign prostatic hyperplasia by suppressing 5α-reductase and androgen receptor expression in rats

doi: 10.4162/nrp.2018.12.5.378

Figure Lengend Snippet: (A) The serum concentrations of 5α-reductase type 2 were examined by ELISA. The data shown represent the mean ± SEM of six rats per group ( P < 0.05). (B) Representative photomicrographs of prostate tissues immunostained with an anti-5α-reductase type 2 are presented (magnification, 200X). Control, corn oil-injected and PBS-treated rats; BHP, TP (3 mg/kg)- and PBS-treated rats; CF-250, TP (3 mg/kg)- and CF (250 mg/kg)-treated rats; CF-500, TP (3 mg/kg)- and CF (500 mg/kg)-treated rats; CF-750, TP (3 mg/kg)- and CF (750 mg/kg)-treated rats; FINA, TP (3 mg/kg)- and finasteride (5 mg/kg)-treated rats. (C and D) The expression levels of 5α-reductase type 2 in prostate tissues were determined by Western blotting. The experiment was repeated three times among different experimental groups, and all the results were similar (C). Actin was used as an internal control. Values are mean ± SD of data from three separate experiments ( P < 0.05) (D). BPH, benign prostatic hyperplasia; CF, Corni Fructus water extract; FINA, finasteride.

Article Snippet: Serum concentrations of testosterone and DHT were determined by using Testosterone enzyme-linked immunosorbent assay (ELISA) kit (BioVision, Inc., Milpitas, CA, USA) and rat DHT ELISA kit (Mybiosource Inc., San Diego, CA, USA), according to the manufacturer's instructions, which are based on the principle of competitive inhibition enzyme immunoassay.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Injection, Expressing, Western Blot

Journal: Nutrition Research and Practice

Article Title: Corni Fructus attenuates testosterone-induced benign prostatic hyperplasia by suppressing 5α-reductase and androgen receptor expression in rats

doi: 10.4162/nrp.2018.12.5.378

Figure Lengend Snippet: (A) The serum concentrations of PSA were examined by ELISA. The data shown represent the mean ± SEM of six rats per group. The *** P < 0.001 vs. Control group; ## P < 0.005, ### P < 0.001 vs. BPH group. (B and C) The expression levels of PSA in prostate tissues were determined by Western blotting. The experiment was repeated three times among different experimental groups, and all the results were similar. Actin was used as an internal control. Values are mean ± SD of data from three separate experiments ( P < 0.05) (C). BPH, benign prostatic hyperplasia; CF, Corni Fructus water extract; FINA, finasteride; PSA, prostate-specific antigen.

Article Snippet: Serum concentrations of testosterone and DHT were determined by using Testosterone enzyme-linked immunosorbent assay (ELISA) kit (BioVision, Inc., Milpitas, CA, USA) and rat DHT ELISA kit (Mybiosource Inc., San Diego, CA, USA), according to the manufacturer's instructions, which are based on the principle of competitive inhibition enzyme immunoassay.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Expressing, Western Blot