Cathepsin B Regulates Mice Granulosa Cells’ Apoptosis and Proliferation In Vitro

Cathepsin B (CTSB), a lysosomal cysteine protease’s high expression and activity, has been reported to cause poor-quality embryos in porcine and bovine. Nevertheless, CTSB functions in mice granulosa cells remain to explore. To discuss the CTSB functional role in follicular dynamics, we studied apoptosis, proliferation, cell cycle progression, and related signaling pathways in primary mouse granulosa cells transfected with small interference RNA specific to CTSB (siCTSB) for 48 h. Further, mRNA and protein expression of cell proliferation regulators (Myc and cyclin D2), apoptosis regulators (caspase 3, caspase 8, TNF-α, and Bcl2), steroidogenesis-related genes (FSHR and CYP11A1), and autophagy markers (LC3-I and ATG5) were investigated. In addition, the effect of CTSB on steroidogenesis and autophagy was also examined. Flow cytometry analysis assay displayed that silencing of CTSB decreased the early and total apoptosis rate by downregulating TNF-α, caspase 8, and caspase 3, and upregulating Bcl2. By regulating Myc and cyclin D2 expression and activating the p-Akt and p-ERK pathways, CTSB knockdown increased GC proliferation and number. A significant decline in estradiol and progesterone concentrations was observed parallel to a significant decrease in autophagy-related markers LC3-I and ATG5 compared to the control group. Herein, we demonstrated that CTSB serves as a proapoptotic agent and plays a critical role in folliculogenesis in female mice by mediating apoptosis, autophagy, proliferation, and steroidogenesis. Hence, CTSB could be a potential prognostic agent for female infertility.

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A Novel Role of SIRT1/ FGF-21 in Taurine Protection Against Cafeteria Diet-Induced Steatohepatitis in Rats

Cellular Physiology and Biochemistry ◽

10.1159/000480649 ◽

2017 ◽

Vol 43 (2) ◽

pp. 644-659 ◽

Cited By ~ 7

Author(s):

Azza H. Abd Elwahab ◽

Basma K. Ramadan ◽

Mona F. Schaalan ◽

Amina M. Tolba

Keyword(s):

Caspase 3 ◽

B Cell Lymphoma ◽

Cafeteria Diet ◽

Control Group ◽

Albino Rats ◽

Protective Mechanism ◽

Alcoholic Fatty Liver ◽

Group I ◽

Tnf Α

Background: Non-alcoholic fatty liver disease (NAFLD) is one of the alarmingly rising clinical problems in the 21st century with no effective drug treatment until now. Taurine is an essential amino acid in humans that proved efficacy as a non-pharmacological therapy in a plethora of diseases; however, its impact on NAFLD remains elusive. The aim of the current study is to evaluate the protective mechanism of taurine in experimental steatohepatitis induced by junk food given as cafeteria-diet (CAF-D) in male albino rats. Methods: Forty adult male albino rats of local strain between 8-10 weeks old, weighing 150 ± 20 g, were divided into four equal groups: Group I (control group), Group II (Taurine group), Group III (CAF-D for 12 weeks) and Group IV (CAF-D +Taurine). CAF-D was given in addition to the standard chow for 12 weeks, where each rat was given one piece of beef burger fried in 15 g of sunflower oil, one teaspoonful of mayonnaise, and one piece of petit pan bread, weighing 60g/ piece. In the serum, liver function tests; ALT, AST, ALP, GGT and the lipid profile; TG, TC, HDL-C added to reduced glutathione (GSH) were assessed colorimetrically, while fibroblast growth factor (FGF)-21, adiponectin & interleukin (IL)-6 via ELISA. The same technique was used for the assays of the hepatic levels of FGF-21, silent information regulator (SIRT1), malondialdehyde (MDA),IL-10, tumor necrosis factor-α (TNF-α) as well as the apoptotic markers; caspase-3 and B-cell lymphoma (Bcl-2). Results: The cafeteria-diet induced steatohepatitis was reflected by significantly increased body and liver weight gain, elevation of liver enzymes; ALT, AST, ALP and GGT added to the dyslipidemic panel, presented as increased TC, TG, LDL-C and decreased HDL-C levels. The steatosis-induced inflammatory milieu, marked by elevated serum levels of FGF-21, IL-6, hepatic TNF-α, as well as reduced IL-10 and adiponectin, was associated with steatosis- induced hepatic oxidative stress, reflected by increased hepatic MDA and decreased GSH levels, along with stimulated caspase-3 and decline in BcL-2 hepatic levels. These pathological disturbances were significantly ameliorated by taurine supplementation and evidenced histopathologically. The cross talk between hepatic FGF-21 and SIRT1 and their association to the induced perturbations are novel findings in this study. Taurine's efficacy in restoration of hepatic structure and function is partially via the increase in SIRT1 and associated reduction of FGF-21. Conclusion: The findings of the current study prove the protective role of taurine in NAFLD via a novel role in the amelioration of FGF-21/ SIRT1 axis, which could be considered a new therapeutic target.

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RNAi inhibition of mineralocorticoid receptors prevents the development of cold-induced hypertension

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.01319.2007 ◽

2008 ◽

Vol 294 (4) ◽

pp. H1880-H1887 ◽

Cited By ~ 22

Author(s):

Zhongjie Sun ◽

Mahajoub Bello-Roufai ◽

Xiuqing Wang

Keyword(s):

Plasma Level ◽

Critical Role ◽

Control Level ◽

Plasma Renin ◽

Control Group ◽

Renal Hypertrophy ◽

Effective Prevention ◽

Small Interference Rna ◽

Exposure To Cold ◽

Induced Hypertension

The objective was to determine whether the mineralocorticoid receptor (MR) plays a role in the initiation and development of cold-induced hypertension (CIH) by testing the hypothesis that the RNA interference (RNAi) inhibition of the MR attenuates CIH. The recombinant adeno-associated virus (AAV) carrying a short-hairpin small-interference RNA for MR (MRshRNA) or a scrambled sequence (ControlshRNA) was constructed. Six groups of albino mice were used (6 mice/group). Three groups were exposed to cold (6.7°C), whereas the remaining three groups were kept at room temperature (RT; warm) as controls. In each temperature condition, three groups received an intravenous injection of MRshRNA, ControlshRNA, or virus-free PBS, respectively, before exposure to cold. The viral complexes (0.35 × 1011 particles/mouse, 0.5 ml) or PBS (0.5 ml) was delivered into the circulation via the tail vein. The blood pressure (BP) of the mice treated with ControlshRNA or PBS increased significantly during exposure to cold, whereas the BP of the cold-exposed MRshRNA-treated mice did not increase and remained at the level of the control group kept at RT. Thus AAV delivery of MRshRNA prevented the initiation of CIH. MRshRNA significantly attenuated cardiac and renal hypertrophy. MRshRNA decreased the cold-induced increase in MR protein expression to the control level in the hypothalamus, kidneys, and heart, indicating an effective prevention of the cold-induced upregulation of MR. RNAi inhibition of MR resulted in significant decreases in the plasma level of norepinephrine, plasma renin activity, and plasma level of aldosterone in cold-exposed mice. MR played a critical role in the initiation and development of CIH. AAV delivery of MRshRNA may serve as a new approach for the prevention of cold-induced hypertension.

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Features of a necrotic and inflammatory process in different forms of nonalcoholic fatty liver disease

Terapevticheskii arkhiv ◽

10.17116/terarkh201789252-58 ◽

2017 ◽

Vol 89 (2) ◽

pp. 52-58 ◽

Cited By ~ 2

Author(s):

I V Kurbatova ◽

O P Dudanova

Keyword(s):

Gene Expression ◽

Liver Disease ◽

Fatty Liver ◽

Proinflammatory Cytokines ◽

Fatty Liver Disease ◽

Inflammatory Process ◽

Caspase 3 ◽

Control Group ◽

Nonalcoholic Fatty Liver ◽

Tnf Α

Aim. To identify the features of development of a necrotic and inflammatory process in different forms of nonalcoholic fatty liver disease (NAFLD), by comparatively analyzing a full set of clinical and laboratory parameters, including the cytokine status and the expression level of enzyme genes controlling the apoptosis of peripheral leukocytes. Subjects and methods 86 patients with NAFLD, including 8 (9.3%) with hepatic steatosis (HS), 70 (81.4%) with nonalcoholic steatohepatitis (NASH), 40, 19, and 11 with mild, moderate, and high disease activity, respectively, and 8 (9.3%) with liver cirrhosis (LC), were examined. A control group consisted of 34 healthy donors. Clinical and biochemical blood indices, cytokine profile, and the level of caspase gene transcripts in the peripheral blood leukocytes (PBL) were estimated. Results. As compared to the controls, the patients with HS had higher tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6) levels and lower caspase 3, 6, and 8 mRNA in PBL. The concentration of IL-10 in NASH was higher than that in steatosis and positively correlated with the level of proinflammatory cytokines. The levels of TNF-α and IL6 were higher in the patients with NASH than in the controls. Those of C-reactive protein, γ-globulin, IL-6, and cytokeratin-18 fragment increased with the progression of NASH. In the latter, the transcriptional activity of caspase-3 gene decreased relative to the reference value and negatively correlated with the level of proinflammatory cytokines. In the patients with LC, the gene expression profile of caspases in PBL was similar to that in the control group; the level of IL-6 was higher than that in steatosis and NASH, that of IL-1β was higher than in HS and positively correlated the concentration of IL-6 and the activity of alanine aminotransferase and aspartate aminotransferase. Conclusion. The features of a necrotic and inflammatory process were identified in different forms of NAFLD. When the latter progressed, the cytokine profile and gene expression levels of caspases in PBL altered along with a change in the general clinical picture.

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TNFR1 is Upregulated and Mediates Apoptosis in Cumulus Cells of Women With Polycystic Ovarian Syndrome

10.21203/rs.3.rs-596490/v1 ◽

2021 ◽

Author(s):

Yaping Jiang ◽

Rui Jiang ◽

Peng Zhang ◽

qiong Yu ◽

Hongping Ba ◽

...

Keyword(s):

Polycystic Ovary Syndrome ◽

Signaling Pathway ◽

Granulosa Cells ◽

Follicular Fluid ◽

Polycystic Ovary ◽

Cumulus Cells ◽

Control Group ◽

Ovary Syndrome ◽

Tnf Α ◽

Pcos Group

Abstract Purpose To investigate the changes of human granulosa cell, TNFR1, TNFR2 and their downstream molecules in patients with polycystic ovary syndrome (PCOS) and the control group. Methods We recruited infertile women with polycystic ovary syndrome (n = 30) and compared them with infertility due to fallopian tube obstruction(n = 30, control group). The ovaries were stimulated with GnRH agonists and gonadotropins. Follicular fluid from large follicles ([14 mm]) was pooled and granulosa cells (GCs) were separated by a cellular filter. The TNF-α level of follicular fluid was measured by ELISA. TUNEL assay were used to detect the apoptosis of purified GCs. Real-time PCR and Western blotting were used to detect the expression of TNF-related signaling molecules in GCs. Results The rate of high quality embryos in the PCOS group was lower than that in the control group. There were higher percentages of apoptosis in GCs of PCOS patients than in the control group. TNF-α is upregulated in follicular fluid of PCOS patients. TNFR1 and caspase-3 mRNA level were signifificantly higher in PCOS group than in the control group. TNF-α-mediated apoptosis of PCOS granulosa cells was mainly dependent on TNFR1.The TNF-α/TNFR1 signaling pathway mediates apoptosis rather than survival in cumulus cells of PCOS patients. Conclusions TNF-α expression was upregulated in follicular fluid of PCOS patients, and TNFR1 overexpression in female granulosa cells of PCOS was associated with higher levels of apoptosis in these cells, suggesting that the TNF-α/TNFR1 signaling pathway may be a candidate for higher apoptosis in female granulosa cells of PCOS.

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MON-027 Activation of Endoplasmic Reticulum Stress Mediates Oxidative Stress-Induced Apoptosis of Granulosa Cells in Ovaries Affected by Endometrioma

Journal of the Endocrine Society ◽

10.1210/jendso/bvaa046.346 ◽

2020 ◽

Vol 4 (Supplement_1) ◽

Author(s):

Chisato Kunitomi ◽

Miyuki Harada ◽

Jerilee Mariam Khong Azahry

Keyword(s):

Oxidative Stress ◽

Er Stress ◽

Granulosa Cells ◽

Caspase 3 ◽

Caspase 8 ◽

Ovarian Dysfunction ◽

High Oxidative Stress ◽

Induced Apoptosis ◽

Sensor Proteins ◽

Apoptotic Factors

Abstract Endometriosis exerts detrimental effects on ovarian physiology and compromises follicular health. Granulosa cells of endometriosis patients are characterized by increased apoptosis, as well as high oxidative stress. Among several pathophysiologic factors associated with endometriosis, it is expected that oxidative stress contributes to the induction of apoptosis in granulosa cells, although the underlying mechanism remains unclear. Endoplasmic reticulum (ER) stress, a local factor closely associated with oxidative stress, has emerged as a critical regulator of ovarian function. We hypothesized that ER stress is activated by high oxidative stress in granulosa cells in ovaries with endometrioma and mediates oxidative stress-induced apoptosis. Ovaries from patients with endometrioma and control were collected to determine apoptosis, oxidative stress and ER stress by TUNEL, immunohistochemical staining of 8-OHdG and ER stress sensors, respectively. Human granulosa-lutein cells (GLCs) obtained from IVF patients were cultured with H2O2 (an oxidative stress inducer) or tauroursodeoxycholic acid (TUDCA, an ER stress inhibitor in clinical use) to assess apoptosis and ER stress by quantitative PCR and FACS. Activity of pro-apoptotic factors was determined by caspase-8 activity assay and western blotting for cleaved caspase-3. Human GLCs from patients with endometrioma expressed up to two times higher level of mRNAs associated with the unfolded protein response (UPR), including ATF4, ATF6, the spliced form of XBP1, HSPA5, and CHOP. In addition, the levels of phosphorylated ER stress sensor proteins, IRE1 and PERK, were elevated. Given that ER stress results in phosphorylation of ER stress sensor proteins and induces UPR factors, these findings indicate that these cells were under ER stress. H2O2 increased expression of UPR-associated mRNAs in cultured human GLCs, and this effect was abrogated by pre-treatment with TUDCA. Treatment with H2O2 increased apoptosis and the activity of pro-apoptotic factors caspase-8 and caspase-3, both of which were attenuated by TUDCA. Our findings suggest that activated ER stress induced by high oxidative stress in granulosa cells in ovaries with endometrioma mediates apoptosis of these cells, leading to ovarian dysfunction in endometriosis patients. Targeting ER stress with currently clinically available ER stress inhibitors, or with these agents in combination with antioxidants, may serve as a novel strategy for rescuing endometriosis-associated ovarian dysfunction.

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Protective Effects of Astaxanthin on Nephrotoxicity in Rats with Induced Renovascular Occlusion

Combinatorial Chemistry & High Throughput Screening ◽

10.2174/1386207323666200914104432 ◽

2020 ◽

Vol 23 ◽

Author(s):

Erkan Arslan ◽

Hakan Turk ◽

Murat Caglayan ◽

Tugba Taskin Turkmenoglu ◽

Ataman Gonel ◽

...

Keyword(s):

Oxidative Stress ◽

Olive Oil ◽

Caspase 3 ◽

Ischemia Reperfusion ◽

Renal Ischemia ◽

Control Group ◽

Tubular Damage ◽

Total Thiol ◽

Tnf Α ◽

Anti Inflammatory

Background: Various effects of Astaxanthin was shown in the studies including its antioxidant, anti-inflammatory, anti-tumor and immunregulator effects. Objective: The aim of this study was to evaluate the beneficial effects of Astaxanthin on renovascular occlusion induced renal injury and to investigate the possible mechanisms. Methods: The rats were randomly assigned into three groups as follows: Group 1: control group (n=12), Group 2: renal ischemiareperfusion injury group (n=12), Group 3: renal ischemia-reperfusion + asthaxantine treated group (n=12). The control group and the renal ischemia-reperfusion group were given 2cc/kg/g olive oil for 7 days before establishing ischemia to renal tissue. Astaxanthin dissolved in olive oil was given orally to the renal ischemia+astaxanthin group for 7 days before inducing renal ischemia. Caspase-(3, 8, 9), GSH, SOD, Total Thiol, TNF-α, IL-6, 8-OHdG were performed for each group. Results: Renal IRI was verified by analysing the pathological changes of renal tissues and the renal functions after renal reperfusion. Much less renal tubular damage was determined the IRI+ASX group in comparison to the IRI group. Caspase-8, -9 and -3 immunoreactivity was observed to be minimal in the control group. Apoptosis was observed to be significantly reduced in the IRI + ASX group with respect to IRI group and close to the level of the control group (p <0.05). Caspase-3 levels of tissue samples were significantly increased in IRI group compared to other groups, but significantly lower in IRI+ASX group with respect to the IRI group (p<0.05). The TOS and OSI levels, indicating increased oxidative stress, were significantly lower in the IRI+ASX group with respect to the IRI group (p <0.001), but still higher than the control group (p <0.001). In addition to GSH, SOD and Total Thiol levels, TAS levels were also significantly higher in IRI + ASX group in comparison to the IRI group (p <0.05). TNF-α, IL-6, lipid hydroperoxide, AOPP and 8-OHdG levels were lower in the IRI+ASX group than the IRI group (p <0.001). MPO, IL-6, TNF-α levels, representing the parameters indicating neutrophil infiltration and inflammation of the renal tissues, significantly increased in IRI group with respect to the other groups (p <0.005). Conclusion: When all the data obtained in our study were evaluated, ASX was determined to prevent renal damage due to renovascular occlusion to a great extent, through complex mechanisms involving antioxidant, anti-inflammatory and antiapopitotic effects. Biochemical, histological and oxidative stress parameters were improved due to ASX.

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P–648 In-vitro Evaluation of SCF and StxBP1 in Polycystic Ovary Syndrome (PCOS)

Human Reproduction ◽

10.1093/humrep/deab130.647 ◽

2021 ◽

Vol 36 (Supplement_1) ◽

Author(s):

S Erol ◽

S Zırh ◽

L Karako. Sokmensuer ◽

G Bozdag ◽

S F Muftuoglu

Keyword(s):

Granulosa Cells ◽

Polycystic Ovary ◽

Critical Role ◽

Vesicle Fusion ◽

Snare Complex ◽

Control Group ◽

Signal Molecules ◽

Expression Levels ◽

Before And After

Abstract Study question Is the interaction between intrafollicular cells in PCOS, impaired by the change of vesicular fusion and/or exocytosis in granulosa cells (GCs)? Summary answer StxBP1 expression leves impared in GCs of PCOS. What is known already PCOS characterised as follicular arrest on antral follicles, cystic follicle formation, and follicular development failure. GCs secretes wide variety of factors via exocytosis, and plays critical role during folliculogenesis. Secretory vesicles are transported to cellular membrane. This process requires local concentrations of SNAREs consisting of tSNARE, vSNARE, and other vesicle fusion associated proteins. SNARE proteins are involved in vesicle fusion, exocytosis, and intracellular trafficking. GCs secretes KITL which provides follicular activation and growth. Syntaxins are one of the members of SNARE complex. StxBP1 is a protein which has a crucial role in secretory vesicle fusion that provides fusion of syntaxins. Study design, size, duration Granulosa cells (GCs) were collected for primary cell culture, since 2019 from both PCOS (n = 10) and healthy (male factor infertility) (n = 10) women undergoing ART. Each GCs from participant divided into two groups as in-vitro stimulated group and in-vitro nonstimulated group. Participants/materials, setting, methods GCs have been isolated from follicular fluid taken from patients during oocyte pick-up at Hacettepe University In-Vitro Fertilization Unit. nGCs were cultured at most secod subcultures after the isolation. The stimulated groups of both PCOS and control groups were stimulated by hCG(10IU/ml) ve FSH(0.5IU/ml) for 24 hours. Vesicle fusion proteins (Stx6, StxBP1, and SNAP25), KITL, and FSHr expressions were analyzed on granulosa cells from each group via immunofluorescent (IF) labeling and cyto-ELISA. Main results and the role of chance FSHr were compared in both control and PCOS before and after stimulation. There was no difference between FSHr expression levels in both groups. Indirect IF is widely considered for SNAP25, Stx6, StxBP1 proteins in all groups of GCs screening with/without stimulation. Expression of SNAP25, StxBP1 mainly scattered through all cytoplasmic area,s and membranous localization was observed. Stx6 expressions were particularly distinguished at perinuclear area of cytoplasm. However, stimulated cells of control appeared more peripherally Stx6 expression. This pattern caused by stimulation wasn’t observed in PCOS. Expressions of SNAP25, Stx6, StxBP1 were observed with less expression in PCOS. Also, the response to stimulation was lower than the control group. The differences in Stx6, SNAP25, StxBP1 and KITL levels before and after stimulation was evaluated for both control and PCOS in Cyto-ELISA. However, both SNAP25 and Stx6 expressions in GCs of both groups were similar in response to stimulation. The expression levels of StxBP1 in response to stimulation were significantly lesser than control at PCOS. KITL expressions were lower in the PCOS as expected furthermore similar to StxBP1 in response to stimulation. According to our findings, the highest response to stimulation in GCs occurred for StxBP1 and KITL in the control. Limitations, reasons for caution Since human cells were used in the study and the cells of each patient do not exhibit the same characteristics, the lowest number of patient samples identified in the statistical power analysis were included in the study. Wider implications of the findings: Our view to the disruption in the secretion of signal molecules in terms of vesicle dynamics will offer a new perspective in female infertility or cross-talks in folliclar cells of the ovary. Trial registration number TSA–2019–18196

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The extrinsic caspase pathway modulates endotoxin-induced diaphragm contractile dysfunction

Journal of Applied Physiology ◽

10.1152/japplphysiol.00377.2006 ◽

2007 ◽

Vol 102 (4) ◽

pp. 1649-1657 ◽

Cited By ~ 19

Author(s):

Gerald S. Supinski ◽

Xinying Ji ◽

Wenyi Wang ◽

Leigh A. Callahan

Keyword(s):

Caspase 3 ◽

Death Receptor ◽

Interferon Γ ◽

Caspase 8 ◽

Contractile Dysfunction ◽

Tnf Receptor ◽

Caspase Activation ◽

Caspase 9 ◽

Diaphragm Dysfunction ◽

Tnf Α

The mechanisms by which infections induce diaphragm dysfunction remain poorly understood. The purpose of this study was to determine which caspase pathways (i.e., the extrinsic, death receptor-linked caspase-8 pathway, and/or the intrinsic, mitochondrial-related caspase-9 pathway) are responsible for endotoxin-induced diaphragm contractile dysfunction. We determined 1) whether endotoxin administration (12 mg/kg IP) to mice induces caspase-8 or -9 activation in the diaphragm; 2) whether administration of a caspase-8 inhibitor ( N-acetyl-Ile-Glu-Thr-Asp-CHO, 3 mg/kg iv) or a caspase-9 inhibitor ( N-acetyl-Leu-Glu-His-Asp-CHO, 3 mg/kg iv) blocks endotoxin-induced diaphragmatic weakness and caspase-3 activation; 3) whether TNF receptor 1-deficient mice have reduced caspase activation and diaphragm dysfunction following endotoxin; and 4) whether cytokines (TNF-α or cytomix, a mixture of TNF-α, interleukin-1β, interferon-γ, and endotoxin) evoke caspase activation in C2C12 myotubes. Endotoxin markedly reduced diaphragm force generation ( P < 0.001) and induced increases in caspase-3 and caspase-8 activity ( P < 0.03), but failed to increase caspase-9. Inhibitors of caspase-8, but not of caspase-9, prevented endotoxin-induced reductions in diaphragm force and caspase-3 activation ( P < 0.01). Mice deficient in TNF receptor 1 also had reduced caspase-8 activation ( P < 0.001) and less contractile dysfunction ( P < 0.01) after endotoxin. Furthermore, incubation of C2C12 cells with either TNF-α or cytomix elicited significant caspase-8 activation. The caspase-8 pathway is strongly activated in the diaphragm following endotoxin and is responsible for caspase-3 activation and diaphragm weakness.

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hnRNP K plays a protective role in TNF-α-induced apoptosis in podocytes

Bioscience Reports ◽

10.1042/bsr20180288 ◽

2018 ◽

Vol 38 (3) ◽

Cited By ~ 1

Author(s):

Shili Zhao ◽

Junxia Feng ◽

Qi Wang ◽

Lu Tian ◽

Yunfang Zhang ◽

...

Keyword(s):

Caspase 3 ◽

Glycogen Synthase ◽

Kidney Diseases ◽

Cell Cycle Distribution ◽

Caspase 8 ◽

G1 Arrest ◽

Inhibitory Protein ◽

Hnrnp K ◽

Tnf Α ◽

Induced Apoptosis

Apoptosis of podocytes contributes to proteinuria in many chronic kidney diseases. The cytokine, tumor necrosis factor-α (TNF-α) is thought to be involved in podocyte apoptosis, but the underlying mechanism is not understood. In our study, we established a model of TNF-α-induced apoptosis by isolating primary podocytes from mice. After exposing cells to TNF-α, we determined the expression levels of heterogeneous nuclear ribonucleoprotein K (hnRNP K) and cellular FLICE-inhibitory protein (c-FLIP) and the phosphorylation levels of glycogen synthase kinase β (GSK3β) and extracellular signal-regulated kinase (ERK). We then knocked down or overexpressed the levels of hnRNP K and observed its effects on the expressions of c-FLIP, caspase-8, caspase-3, and the phosphorylation of GSK3β and ERK. In addition, we examined the percentage of cells undergoing apoptosis and studied cell cycle distribution. We found that TNF-α induced apoptosis in podocytes and that the expressions of hnRNP K and c-FLIP were significantly decreased, whereas the phosphorylations of GSK3β and ERK were significantly increased. Both gene knockdown and overexpression of hnRPN K resulted in varied expressions/phosphorylations of c-FLIP, GSK3β, and ERK. Moreover, decreased hnRPN K expression contributed to increased levels of caspase-8 and capase-3, as well as an increase in cell apoptosis and G0/G1 arrest. In conclusion, down-regulated expression of hnRNP K by TNF-α resulted in a decrease in the expression of c-FLIP as well as increases in phosphorylated GSK3β, ERK, caspase-8, and caspase-3, and then critically contributed to the podocyte apoptosis.

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Catechins decrease neurological severity score through apoptosis and neurotropic factor pathway in rat traumatic brain injury

Zinc supplementation improves heme biosynthesis in rats exposed to lead ◽

10.18051/univmed.2017.v36.110-122 ◽

2017 ◽

Vol 36 (2) ◽

pp. 110

Author(s):

Retty Ratnawati ◽

Annisa Nurul Arofah ◽

Anastasia Novitasari ◽

Sartika Dewi Utami ◽

Made Ayu Hariningsih ◽

...

Keyword(s):

Traumatic Brain Injury ◽

Brain Injury ◽

Severity Score ◽

Caspase 3 ◽

Negative Control Group ◽

Positive Control ◽

Negative Control ◽

Control Group ◽

Caspase 8 ◽

Neurotropic Factor

BACKGROUND Catechins inhibits apoptosis through anti oxidant and anti inflamation pathway. Catechins also increases brain-derived neurotrophic factor (BDNF). There was a few research that explained the role of catechins in traumatic brain injury (TBI). The objective of the study was to evaluate the effect of catechins administration on neurologic severity score (NSS) through apoptosis and neurotropic pathway in traumatic brain injury rat model.METHODS A post test only controlled group design was performed using traumatic brain injury rat (Rattus novergicus) model through weight drop models. It was devided into negative control group, positive control group, TBI+catechins 513 mg/kgBW, TBI+catechins 926 mg/kgBW, TBI+catechins 1113 mg/kgBW. NSS was measured in the first hours, day three, and day seven. The expressions of NFkB, TNFa, Bcl-2, Bax, caspase 3, caspase 8, BDNF, and the numbers of apoptosis cells were evaluated by imunohistochemystry method. One way Anova and Kruskal Wwallis were used to analyse the data.Results TNFa, caspase 8, number of apoptosis cells were significantly decreased on the seventh day administration compared to the third day administration (p<0.05). Catechins increased the expression of Bcl-2/Bax and BDNF significantly (p<0.05). Yet, there were no significant differences between expression of caspase 3, NSS, Bcl-2/Bax ratio, and BDNF toward third days administration of catechins compared with seven days administration (p>0.050).Conclusions Administration of catechins decreased NSS through inhibiting inflammation and apoptosis, as well as induced the neurotrophic factors in rat brain injury. Catechins may serve as a potential intervention for TBI.

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