scholarly journals Down regulating PHGDH affects the lactate production of Sertoli cells in varicocele

2020 ◽  
Author(s):  
Wen-bin Guo ◽  
Zhen-hui Huang ◽  
Cheng Yang ◽  
Xian-yuan Lv ◽  
Hui Xia ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Male Infertility ◽  
Western Blot ◽  
Sertoli Cells ◽  
Venous Blood ◽  
Lactate Concentration ◽  
Lactate Production ◽  
Glucose Consumption ◽  
Down Regulation ◽  
Regulation Model

Abstract Background: Although varicocele is considered to be one of the leading causes of male infertility, the precise mechanism underlying how varicocele leads to male infertility is not completely understood. We found the lactate concentration on the varicocele side of the patients was decreased compare with peripheral venous blood. In the testicles, the lactate produced by the sertoli cells (TM4) through the glycolysis pathway provides most of the energy needed for spermatogenesis, the reduction of lactate will affect spermatogenesis. The objective of this study was to investigate the mechanism of this abnormal energy metabolism phenomenon in varicocele.Methods: In this study, we collected the testicular tissue from patients with varicocele, the glycolysis related proteins PHGDH was identified by iTRAQ proteomics technology. Experimental rat varicocele model was constructed according to our new clip technique, the mRNA and protein expression levels of PHGDH were examined with qRT-PCR and Western blotting. We constructed a TM4 cell of PHGDH down-regulation model, and then detected the glucose consumption, LDH activities and lactate production in the TM4 cells. Western blot was conducted to investigate the effects of PHGDH on the expression of phosphoserine phosphatase (PSPH) and Pyruvate kinase M2 (PKM2). Flow cytometry was used to detect the cell apoptosis and cell cycle in TM4 cells.Results: The results showed that testicular protein PHGDH was down-regulated in varicocele. Down-regulation of PHGDH in TM4 cells significantly decreased the glucose consumption, LDH activities and lactate production in the TM4 cells, indicating that the low expression of PHGDH ultimately led to a decrease in lactate production by affecting the glycolysis. The Western blot results showed that the down-regulation of PHGDH significantly reduced the expression of pathway protein PSPH and PKM2, leading to the reduction of lactate production. Moreover, PHGDH knockdown can promote apoptosis and inhibit cell cycle to affect cell growth.Conclusions: Overall, we conformed that varicocele lead to the decreasing of testis lactate production. Down-regulation of PHGDH in TM4 cells may mediate the process of abnormal glucose metabolism. Our study provide new insight into the mechanisms underlying metabolism-associated male infertility and suggests a novel therapeutic target for male infertility.

scholarly journals Down regulating PHGDH affects the lactate production of Sertoli cells in varicocele

2020 ◽  
Author(s):  
Wen-bin Guo ◽  
Zhen-hui Huang ◽  
Cheng Yang ◽  
Xian-yuan Lv ◽  
Hui Xia ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Male Infertility ◽  
Western Blot ◽  
Sertoli Cells ◽  
Venous Blood ◽  
Lactate Concentration ◽  
Lactate Production ◽  
Glucose Consumption ◽  
Down Regulation ◽  
Regulation Model

Abstract Background: Although varicocele is considered to be one of the leading causes of male infertility, the precise mechanism underlying how varicocele leads to male infertility is not completely understood. We found the lactate concentration on the varicocele side of the patients was decreased compare with peripheral venous blood. In the testicles, the lactate produced by the sertoli cells through the glycolysis pathway provides most of the energy needed for spermatogenesis, the reduction of lactate will affect spermatogenesis. The objective of this study was to investigate the mechanism of this abnormal energy metabolism phenomenon in varicocele.Methods: In this study, we collected the testicular tissue from patients with varicocele, the glycolysis related proteins PHGDH was identified by iTRAQ proteomics technology. Experimental rat varicocele model was constructed according to our new clip technique, the mRNA and protein expression levels of PHGDH were examined with qRT-PCR and Western blotting. We constructed a sertoli cell of PHGDH down-regulation model, and then detected the glucose consumption, LDH activities and lactate production in the sertoli cells. Western blot was conducted to investigate the effects of PHGDH on the expression of phosphoserine phosphatase (PSPH) and Pyruvate kinase M2 (PKM2). Flow cytometry was used to detect the cell apoptosis and cell cycle in sertoli cells.Results: The results showed that testicular protein PHGDH was down-regulated in patients with varicocele and in experimental rat varicocele model. Down-regulation of PHGDH in sertoli cells significantly decreased the glucose consumption, LDH activities and lactate production in the sertoli cells, indicating that the low expression of PHGDH ultimately led to a decrease in lactate production by affecting the glycolysis. The Western blot results showed that the down-regulation of PHGDH significantly reduced the expression of pathway protein PSPH and PKM2, leading to the reduction of lactate production. Moreover, PHGDH knockdown can promote apoptosis and inhibit cell cycle to affect cell growth.Conclusions: Overall, we conformed that varicocele lead to the decreasing of testis lactate production. Down-regulation of PHGDH in sertoli cells may mediate the process of abnormal glucose metabolism. Our study provide new insight into the mechanisms underlying metabolism-associated male infertility and suggests a novel therapeutic target for male infertility.

Costal diaphragmatic O2 and lactate extraction in laryngeal hemiplegic ponies during exercise

1988 ◽  
Vol 65 (4) ◽  
pp. 1723-1728 ◽  
Author(s):  
M. Manohar ◽  
T. E. Goetz ◽  
D. Nganwa
Keyword(s):  
Maximal Exercise ◽  
Venous Blood ◽  
Lactate Concentration ◽  
Lactate Production ◽  
Hemoglobin Concentration ◽  
Insignificant Change ◽  
Resistive Breathing ◽  
O2 Extraction ◽  
Before And After ◽  
Arterial Po2

Diaphragmatic O2 and lactate extraction were examined in seven healthy ponies during maximal exercise (ME) carried out without, as well as with, inspiratory resistive breathing. Arterial and diaphragmatic venous blood were sampled simultaneously at rest and at 30-s intervals during the 4 min of ME. Experiments were carried out before and after left laryngeal hemiplegia (LH) was produced. During ME, normal ponies exhibited hypocapnia, hemoconcentration, and a decrease in arterial PO2 (PaO2) with insignificant change in O2 saturation. In LH ponies, PaO2 and O2 saturation decreased well below that in normal ponies, but because of higher hemoglobin concentration, arterial O2 content exceeded that in normal ponies. Because of their high PaCO2 during ME, acidosis was more pronounced in LH animals despite similar lactate values. Diaphragmatic venous PO2 and O2 saturation decreased with ME to 15.5 +/- 0.9 Torr and 18 +/- 0.5%, respectively, at 120 s of exercise in normal ponies. In LH ponies, corresponding values were significantly less: 12.4 +/- 1.3 Torr and 15.5 +/- 0.7% at 120 s and 9.8 +/- 1.4 Torr and 14.3 +/- 0.6% at 240 s of ME. Mean phrenic O2 extraction plateaued at 81 and 83% in normal and LH animals, respectively. Significant differences in lactate concentration between arterial and phrenic-venous blood were not observed during ME. It is concluded that PO2 and O2 saturation in the phrenic-venous blood of normal ponies do not reach their lowest possible values even during ME. Also, the healthy equine diaphragm, even with the added stress of inspiratory resistive breathing, did not engage in net lactate production.

scholarly journals Increased Pentose Phosphate Pathway Flux after Clinical Traumatic Brain Injury: A [1,2-13C2]glucose Labeling Study in Humans

2007 ◽  
Vol 27 (9) ◽  
pp. 1593-1602 ◽  
Author(s):  
Joshua R Dusick ◽  
Thomas C Glenn ◽  
WN Paul Lee ◽  
Paul M Vespa ◽  
Daniel F Kelly ◽  
...  
Keyword(s):  
Traumatic Brain Injury ◽  
Brain Injury ◽  
Venous Blood ◽  
Lactate Concentration ◽  
Lactate Production ◽  
Jugular Bulb ◽  
Pentose Phosphate ◽  
Severe Tbi ◽  
The Difference ◽  
Gas Chromatography Mass Spectroscopy

Patients with traumatic brain injury (TBI) routinely exhibit cerebral glucose uptake in excess of that expected by the low levels of oxygen consumption and lactate production. This brings into question the metabolic fate of glucose. Prior studies have shown increased flux through the pentose phosphate cycle (PPC) during cellular stress. This study assessed the PPC after TBI in humans. [1,2-13C2]glucose was infused for 60 mins in six consented, severe-TBI patients (GCS < 9) and six control subjects. Arterial and jugular bulb blood sampled during infusion was analyzed for 13C-labeled isotopomers of lactate by gas chromatography/mass spectroscopy. The product of lactate concentration and fractional abundance of isotopomers was used to determine blood concentration of each isotopomer. The difference of jugular and arterial concentrations determined cerebral contribution. The formula PPC = ( m1/ m2)/(3 + ( m1/ m2)) was used to calculate PPC flux relative to glycolysis. There was enrichment of [1,2-13C2]glucose in arterial-venous blood (enrichment averaged 16.6% in TBI subjects and 28.2% in controls) and incorporation of 13C-label into lactate, showing metabolism of labeled substrate. The PPC was increased in TBI patients relative to controls (19.6 versus 6.9%, respectively; P = 0.002) and was excellent for distinguishing the groups (AUC = 0.944, P < 0.0001). No correlations were found between PPC and other clinical parameters, although PPC was highest in patients studied within 48 h of injury (averaging 33% versus 13% in others; P = 0.0006). This elevation in the PPC in the acute period after severe TBI likely represents a shunting of substrate into alternative biochemical pathways that may be critical for preventing secondary injury and initiating recovery.

COMPUTATION OF THE DYNAMIC COMPRESSION EFFECTS IN SPINE DISCS USING INTEGRAL METHODS

2018 ◽  
Vol 18 (05) ◽  
pp. 1750103
Author(s):  
M. CERROLAZA ◽  
F. NIETO ◽  
Y. GONZÁLEZ
Keyword(s):  
Dynamic Loading ◽  
Dynamic Compression ◽  
Lactate Concentration ◽  
Lactate Production ◽  
Glucose Consumption ◽  
Integral Methods ◽  
Boundary Model ◽  
Compression Effects ◽  
Region Approach ◽  
Good Agreement

The computational modeling using integral methods of dynamic loading and its effects on the nutrients transport in spine discs is addressed in this work. The numerical simulation and analysis was carried out using the Boundary Element Method (BEM) and a 3D model (axisymmetric) of the disc. The boundary model was discretized using linear interpolated elements and a multi-region approach. Concentration and production of three nutrients as lactate, oxygen and glucose were obtained. The maximum lactate concentration was observed very close to the interface between the nucleus and the inner annulus. A relatively simple model discretized with 130 boundary elements yielded very similar results to these coming from more complex FEM-based models. The numerical efforts in the domain and boundary discretizations were optimized using the BEM. Our results are in good agreement with those obtained using with finite element-based models. As expected, the dynamic loading increased the oxygen–glucose consumption and the lactate production, thus leading to a poor oxygen–glucose concentration at the nucleus of the disc. All of that is a favorable environment for a disc degeneration mechanism to be developed.

scholarly journals GLUCOSE METABOLISM OF THE SWIMBLADDER TISSUE OF THE EUROPEAN EEL ANGUILLA ANGUILLA

1993 ◽  
Vol 185 (1) ◽  
pp. 169-178 ◽  
Author(s):  
B. Pelster ◽  
P. Scheid
Keyword(s):  
Glucose Uptake ◽  
Venous Blood ◽  
Lactate Concentration ◽  
Lactate Production ◽  
Anguilla Anguilla ◽  
Pentose Phosphate ◽  
Co2 Production ◽  
European Eel ◽  
O2 Uptake ◽  
Lactate Release

Glucose uptake from, and lactate release into, the blood have been analysed in the active gas- depositing swimbladder of the immobilized European eel Anguilla anguilla. Under normoxic conditions, 0.72 micromole min-1 glucose was removed from the blood supply, while lactate was released into it at a rate of 1.16 micromole min-1. The rate of gas deposition into the swimbladder was significantly correlated with the rate of lactate production. Under hypoxic conditions, glucose consumption by, and lactate production of, the swimbladder tissue were reduced, as was the rate of gas deposition. Compared with normoxic conditions, lactate concentration in the swimbladder tissue was elevated after 1 h of hypoxia, indicating a decrease in lactate release. No difference in the osmolality of arterial and venous blood could be detected in these experiments. Combining the data for glucose uptake and lactate release measured under normoxic conditions with the values for O2 uptake and CO2 production of the swimbladder tissue measured under similar conditions in a previous study, a quantitative evaluation of glucose catabolism was performed. According to the O2 uptake of the tissue, only about 1 % of the glucose was oxidized, while about 80 % was fermented to lactic acid. The remaining 0.14 micromole min-1 glucose was presumably catabolized through the pentose phosphate shunt, as indicated by the CO2 production of 0.16 micromole min-1 that cannot be explained by aerobic metabolism.

scholarly journals LncRNA SNHG5 promotes the glycolysis and proliferation of breast cancer cell through regulating BACH1 via targeting miR-299

Breast Cancer ◽  
2021 ◽  
Author(s):  
Shu-Lin Huang ◽  
Zhong-Cheng Huang ◽  
Chao-Jie Zhang ◽  
Jing Xie ◽  
Shan-Shan Lei ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cell Cycle ◽  
Malignant Tumors ◽  
Lactate Production ◽  
Glucose Consumption ◽  
Luciferase Reporter ◽  
Inhibitory Effects ◽  
Glucose Assay ◽  
Non Coding Rna ◽  
Long Non Coding Rna

Abstract Background Breast cancer (BC) is one of the most common malignant tumors in women. Accumulating studies have been reported that long non-coding RNA (lncRNA) SNHG5 is highly expressed in BC. However, the specific molecular mechanism of SNHG5 in BC is unclear. Methods Gene and protein expressions in BC cell were detected by qRT-PCR and western blotting. The proliferation and cell cycle were measured using colony formation assay and flow cytometry analysis, separately. The glucose consumption and lactate production were determined by using the glucose assay kit and lactate assay kit. A dual-luciferase reporter assay was performed to measure the interaction between miR-299 and SNHG5 or BACH1. Results SNHG5 and BACH1 expressions were increased in BC cell while miR-299 level was decreased. SNHG5 increased BACH1 expression by directly targeting miR-299. SNHG5 silencing or miR-299 overexpression suppressed the proliferation of BC cell, arrested the cell cycle in the G1 cell phase, and decreased the glucose consumption and lactate production of BC cell. However, inhibition of miR-299 or overexpression of BACH1 could reverse the inhibitory effects of sh-SNHG5 on cell proliferation and glycolysis in BC. Conclusion SNHG5 promoted the BC cell growth and glycolysis through up-regulating BACH1 expression via targeting miR-299. These findings may improve the diagnostic and therapeutic approaches to BC.

circPUM1 Activates the PI3K/AKT Signaling Pathway by Sponging to Promote to Promote the Proliferation, Invasion and Glycolysis of Pancreatic Cancer

2021 ◽  
Vol 22 ◽  
Author(s):  
Yang Chen ◽  
Yuhong Liu ◽  
Qiang Tao ◽  
YouWen Fan ◽  
Chao Ma ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Cell Proliferation ◽  
Signaling Pathway ◽  
Lactate Production ◽  
Glucose Consumption ◽  
Akt Signaling ◽  
Luciferase Assay ◽  
Akt Signaling Pathway ◽  
Down Regulation ◽  
Apoptosis Rate

Background: Our study seeks to obtain data to assess the impacts of circPUM1 on pancreatic cancer (PC) and its mechanism. Methods: The expression of circPUM1 and miR-200c-3p in PC and normal tissues and PC cell lines was collected and detected. Subsequently, dual-luciferase assay-based verification of the binding site of the two was carried out. After interfering with circPUM1 expression in MIAPaCa-2 and PANC-1 cells, cell proliferation, viability, apoptosis rate, invasion ability, glucose consumption, and lactate production were measured by MTT, colony formation, flow cytometry, Transwell assays, and glucose and lactate assay kits. Additionally, western blot was utilized for assessing PI3K/AKT signaling pathway-related proteins. From the results, highly expressed circPUM1 and miR-200c-3p in PC tissues and cells were proved. Results: Down-regulation of circPUM1 expression significantly inhibited cell proliferation, cell viability, invasion, and glycolysis, while increasing the apoptosis rate. Down-regulated circPUM1 led to the inhibition of the PI3K/AKT signaling pathway activity in PC cells, while up-regulated circPUM1 increased its activity. Further experiments revealed that down-regulation of miR-200c-3p expression reversed the inhibitory effect of lowly expressed circPUM1 on PC cells. Conclusion: In summary, circPUM1 activates PI3K/AKT signaling pathway by sponging miR-200c-3p and promotes PC progression.

scholarly journals Down-regulation of CCNE1 expression suppresses cell proliferation and sensitizes gastric carcinoma cells to Cisplatin

2019 ◽  
Vol 39 (6) ◽  
Author(s):  
Chao Zhang ◽  
Qiang Zhu ◽  
Jianzhong Gu ◽  
Shan Chen ◽  
Qian Li ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Gastric Carcinoma ◽  
Western Blot ◽  
Cell Lines ◽  
Survival Curve ◽  
Lymphatic Invasion ◽  
Cell Counting ◽  
Down Regulation ◽  
Qrt Pcr

AbstractA novel oncogene CCNE1 (cyclin E) is considered to be associated with the development of various tumor types, its role in gastric carcinoma (GC) is little studied and the effect of CCNE1 on chemotherapy also remains unclear. We recruited 55 cases of GC tissues and corresponding normal tissues. Immunohistochemistry (IHC), quantitative real-time PCR (qRT-PCR) and Western blot analysis were performed to detect the expression of CCNE1. We also examined the expression of CCNE1 in gastric mucosal GES-1 cells and five GC cell lines. Silencing CCNE1 was used to assess its effect on proliferation and cell cycle in MGC-803 and NCI-N87 cells, as performed by Cell counting kit-8 (CCK-8) and flow cytometry assay. Meanwhile, cell cycle related genes were also detected through qRT-PCR and Western blot. The results showed CCNE1 up-regulation mainly expressed in GC tissues and GC cell lines, also was associated with tumor node metastasis (TNM) stage and lymphatic invasion. Three-year survival curve analysis showed CCNE1 with high expression had a poor prognosis. Silencing CCNE1 significantly reduced cell viability in 48 h, cultured and arrested cell cycle in G1 phase, moreover, Cyclin A, D1 and C-myc all revealed down-regulation in both MGC-803 and NCI-N87 cells. CCNE1 expression was significantly increased at low and moderate concentrations of Cisplatin. Down-regulation of CCNE1 expression would remarkably promote cell apoptosis induced by Cisplatin, and regulate the rate of Bax/Bcl-2. Down-regulation of CCNE1 expression could inhibit cell proliferation and enhance GC cells sensibility to Cisplatin, possibly involving the regulation of Bcl-2 family.

scholarly journals Circ_0013359 facilitates the tumorigenicity of melanoma by regulating miR-136-5p/RAB9A axis

2021 ◽  
Vol 16 (1) ◽  
pp. 482-494
Author(s):  
Qi Zhang ◽  
Yingfa Feng ◽  
Jiangang Feng ◽  
Jinming Zhang ◽  
Lili Huang
Keyword(s):  
Tumor Growth ◽  
Western Blot ◽  
Lactate Production ◽  
Glucose Consumption ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Circular Rnas ◽  
Melanoma Progression ◽  
Cycle Arrest

Abstract Background Circular RNAs play crucial roles in tumor occurrence and progression. This research aimed to explore the role and potential mechanism of hsa_circ_0013359 (circ_0013359) in melanoma. Methods The levels of circ_0013359, microRNA-136-5p (miR-136-5p), and member RAS oncogene family (RAB9A) in melanoma tissues and cells were detected using quantitative reverse transcriptase-polymerase chain reaction or western blot. Cell proliferation, apoptosis, cell cycle, cell migration, and invasion were evaluated by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay, colony formation assay, flow cytometry, and transwell assay. Glycolysis was determined by detecting glucose consumption, lactate production, and extracellular acidification rate. The levels of hexokinase 2 and lactate dehydrogenase A were examined by western blot. The targeting relationship between miR-136-5p and circ_0013359 or RAB9A was confirmed by dual-luciferase reporter assay. Xenograft experiments were used to analyze tumor growth in vivo. Results Circ_0013359 and RAB9A levels were increased, while the miR-136-5p level was reduced in melanoma tissues and cells. Circ_0013359 knockdown inhibited proliferation, migration, invasion, and glycolysis and promoted apoptosis and cycle arrest in A875 and SK-MEL-1 cells. Circ_0013359 sponged miR-136-5p to regulate melanoma progression. In addition, miR-136-5p suppressed melanoma progression by targeting RAB9A. Besides, circ_0013359 silencing inhibited tumor growth in vivo. Conclusion Depletion of circ_0013359 hindered melanoma progression by regulating miR-136-5p/RAB9A axis.

ÉTUDE IN VITRO DU MÉTABOLISME DES HYDRATES DE CARBONE DANS LE LEUCOCYTE CIRCULANT DU COBAYE TRAITÉ À LA CORTISONE

1966 ◽  
Vol 51 (2) ◽  
pp. 193-202
Author(s):  
J. A. Antonioli ◽  
A. Vannotti
Keyword(s):  
Glucose Concentration ◽  
Intramuscular Injection ◽  
Acute Inflammation ◽  
Glycogen Content ◽  
Glycogen Synthesis ◽  
Lactate Production ◽  
Glucose Consumption ◽  
List Type ◽  
Hydrates De Carbone

ABSTRACT 1. The metabolism of suspensions of circulating leucocytes has been studied after intramuscular injection of a dose of 50 mg/kg of a corticosteroid (cortisone acetate). The suspensions were incubated under aerobic conditions in the presence of a glucose concentration of 5.6 mm. Glucose consumption, lactate production, and variations in intracellular glycogen concentration were measured. After the administration of the corticosteroid, the anabolic processes of granulocyte metabolism were reversibly stimulated. Glucose consumption and lactate production increased 12 hours after the injection, but tended to normalize after 24 hours. The glycogen content of the granulocytes was enhanced, and glycogen synthesis during the course of the incubation was greatly stimulated. The action of the administered corticosteroid is more prolonged in females than in males. The injection of the corticosteroid caused metabolic modifications which resemble in their modulations and in their chronological development those found in circulating granulocytes of guinea-pigs suffering from sterile peritonitis. These results suggest, therefore, that, in the case of acute inflammation, the glucocorticosteroids may play an important role in the regulation of the metabolism of the blood leucocytes.

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