Caspase-3 Protein Expression Research Articles

The Effect of High Intensity Interval Training on the Intrinsic and Extrinsic Pathways of Myocardial Apoptosis in Diet-Induced Obese Rats

Background: Diet-induced obesity is associated with several cardiovascular diseases and myocardial apoptosis. This study aimed to investigate the effect of high intensity interval training (HIIT) on the myocardial apoptosis in the diet-induced obese rats. Methods: Thirty-two male rats, after an obesity induction period, were randomly selected and assigned to four groups including non-obese control (NOC; n=8), basal obese control (BOC; n=8), obese control (OC; n=8), and obese+HIIT (OT; n=8). NOC and BOC groups were sacrificed before the training period. The OT group underwent a HIIT program performed 5 times/week over 12 weeks. Rat hearts were removed 48 hours after the last training session. The Bax, Bcl-2, cytochrome-c, Bid, caspase-8, as well as caspase-3 protein expressions were analyzed using western blotting. Data were analyzed using independent t test. Results: After obesity induction period, the Bax, Bid, cytochrome-c, caspase-8, caspase-3 proteins, and Bax/Bcl-2 in BOC were found to be significantly higher than those in NOC (P=0.025, P=0.0001, P=0.013, P=0.017, P=0.01, P=0.18, respectively). However, Bcl-2 protein in BOC was detected to be significantly lower than that in NOC (P=0.025). The results obtained after completing HIIT training showed that Bid, cytochrome-c, caspase-8, caspase-3, and Bax/Bcl-2 in OT were lower than those in OC (P=0.005, P=0.039, P=0.001, P=0.04, P=0.05, respectively). However, Bcl-2 protein in OT was significantly higher than that in OC (P=0.004). No significant difference was found between OT and OC regarding Bax protein (P=0.32). Conclusion: Diet-induced obesity may have exacerbated the myocardial apoptosis through both intrinsic and extrinsic pathways. However, it seemed that HIIT training significantly prevented the increase of myocardial apoptosis in obese rats.

Effect of Yanshiqiangxin Decoction on Cardiomyocyte Apoptosis in Heart Failure Rats Based on PI3K/Akt/Caspase-9 Signaling Pathway

<bold>Objective</bold> To detect the effect of Yanshiqiangxin decoction on apoptosis of cardiomyocytes in rats with heart failure based on PI3K/Akt/Caspase-9 signalling pathway and to explore the possible mechanism of Yanshiqiangxin decoction in the treatment of heart failure. <bold>Methods</bold> A total of 60 Wistar male rats were randomly divided into the normal group, the model group, the low dose Yanshiqiangxin decoction group, the medium dose Yanshiqiangxin decoction group,the high dose Yanshiqiangxin decoction group and the Perindopril group according to the random number table method, with 10 cases in each group. Except for the normal group, the remaining five groups were modelled by injecting adriamycin solution to induce myocardial injury and to establish the rat model of heart failure. In the low, medium and high dose Yanshiqiangxin decoction groups, 4.325, 8.65 and 17.3 g/(kg·d) of raw drug were administered by gavage respectively; the Perindopril group was given perindopril solution by gavage at the dose of 4 mg/(kg·d). The gavage dose was calculated as 10 mL/(kg·d) ,the normal group and the model group were given the same volume of distilled water. One day after the last gavage, M-mode echocardiography [left ventricular ejection fraction (LVEF), left ventricular short-axis shortening (LVFS), left ventricular internal dimension diastole (LVIDd) and left ventricular internal dimension systole (LVIDs)] was performed through an ultra-high resolution small animal ultrasound imaging system; B-type natriuretic peptide (BNP) was measured by ELISA method; the apoptosis of cardiomyocytes was observed through laser confocal microscopy and the apoptosis index was calculated; the expression of apoptosis-related proteins Bax, Bcl-2 and PI3K/Akt/Caspase-9 signalling pathway proteins were detected by Western blot method. <bold>Results</bold> ① Cardiac ultrasound test results: compared with the normal group, the left ventricular anterior wall motion waveform of the model group was not obvious, close to linear motion, and LVEF and LVFS decreased significantly (<italic>P</italic>&lt;0.05), LVIDd and LVIDs increased significantly (<italic>P</italic>&lt;0.05), the left ventricle was significantly enlarged and the contractility was reduced. Compared with the model group, the amplitude of left ventricular anterior wall motion waveform increased at different degrees in the low, medium and high dose Yanshiqiangxin decoction group, LVEF and LVFS gradually increased with the increase of administration concentration (<italic>P</italic>&lt;0.05), while LVIDd decreased only in the medium dose Yanshiqiangxin decoction group (<italic>P</italic>&lt;0.05), LVIDs of the medium dose Yanshiqiangxin decoction group and the high dose Yanshiqiangxin decoction group were significantly reduced, the medium dose Yanshiqiangxin decoction group decreased the most significantly (<italic>P</italic>&lt;0.05). ② Serum BNP results: compared with the normal group, BNP of the model group was significantly increased (<italic>P</italic>&lt;0.05). Compared with the model group, BNP of the low, medium and high dose Yanshiqiangxin decoction group was significantly decreased, BNP gradually decreased with the increase of administration concentration (<italic>P</italic>&lt;0.05). ③ Myocardial histopathology results: Cardiomyocytes had no apoptosis and almost no positive signal was detected in the normal group; compared with the model group,the number of cardiomyocyte apoptosis of the low, medium, high dose Yanshiqiangxin decoction group and the perindopril group was significantly decreased, the number of cardiomyocyte apoptosis gradually decreased with the increase of administration concentration (<italic>P</italic>&lt;0.05). ④ PI3K/Akt/Caspase-9 signaling pathway protein expression levels and apoptotic factors: compared with the model group, PI3K and Akt protein expression, Bcl-2 content and Bcl-2/Bax ratio of the low, medium and high dose Yanshiqiangxin decoction group were significantly increased, PI3K and Akt protein expression, Bcl-2 content and Bcl-2/Bax ratio gradually increased with the increase of administration concentration (<italic>P</italic>&lt;0.05); Caspase-9 and Caspase-3 protein expression of the low, medium and high dose Yanshiqiangxin decoction group were significantly decreased, Bax content of the medium and high dose Yanshiqiangxin decoction group were significantly decreased, and gradually increased with the increase of administration concentration (<italic>P</italic>&lt;0.05). <bold>Conclusions</bold> Yanshiqiangxin decoction can improve cardiac function, reduce the content of BNP in serum and cardiomyocyte apoptosis, the mechanism may be related to the regulation of the PI3K/Akt/Caspase-9 signalling pathway.

Neuroprotective effect of liraglutide in an experimental mouse model of multiple sclerosis: role of AMPK/SIRT1 signaling and NLRP3 inflammasome

The heterogeneous nature of multiple sclerosis (MS) and the unavailability of treatments addressing its intricate network and reversing the disease state is yet an area that needs to be elucidated. Liraglutide, a glucagon-like peptide-1 analogue, recently exhibited intriguing potential neuroprotective effects. The currents study investigated its potential effect against mouse model of MS and the possible underlying mechanisms. Demyelination was induced in C57Bl/6 mice by cuprizone (400 mg/kg/day p.o.) for 5 weeks. Animals received either liraglutide (25 nmol/kg/day i.p.) or dorsomorphin, an AMPK inhibitor, (2.5 mg/Kg i.p.) 30 min before the liraglutide dose, for 4 weeks (starting from the second week). Liraglutide improved the behavioral profile in cuprizone-treated mice. Furthermore, it induced the re-myelination process through stimulating oligodendrocyte progenitor cells differentiation via Olig2 transcription activation, reflected by increased myelin basic protein and myelinated nerve fiber percentage. Liraglutide elevated the protein content of p-AMPK and SIRT1, in addition to the autophagy proteins Beclin-1 and LC3B. Liraglutide halted cellular damage as manifested by reduced HMGB1 protein and consequently TLR-4 downregulation, coupled with a decrease in NF-κB. Liraglutide also suppressed NLRP3 transcription. Dorsomorphin pre-administration indicated a possible interplay between AMPK/SIRT1 and NLRP3 inflammasome activation as it partially reversed liraglutide’s effects. Immunohistochemical examination of Iba+ microglia emphasized these findings. In conclusion, liraglutide exerts neuroprotection against cuprizone-induced demyelination via anti-inflammatory, autophagic flux activation, NLRP3 inflammasome suppression, and anti-apoptotic mechanisms, possibly mediated, at least in part, via AMPK/SIRT1, autophagy, TLR-4/ NF-κB/NLRP3 signaling.Graphical abstractThe potential mechanistic insight of Lira in alleviating Cup-induced neurotoxicity via: (1) AMPK/SIRT1 pathways activation resulting in the stimulation of brain autophagy flux (confirmed by lowering Beclin-1 and LC3-B protein expression). (2) Inhibition of NLRP3 inflammasome activation, as evidenced by reduced HMGB1, TLR-4, NF-κB and NLRP3 protein expression, alongside diminishing the activation of its downstream cascade as reflected by reduced levels of caspase-1 and IL-1β protein expression. (3) A possible modulating interplay between the previously mentioned two pathways.

Fucoxanthin Prevents Long-Term Administration l-DOPA-Induced Neurotoxicity through the ERK/JNK-c-Jun System in 6-OHDA-Lesioned Mice and PC12 Cells.

As the most abundant marine carotenoid extracted from seaweeds, fucoxanthin is considered to have neuroprotective activity via its excellent antioxidant properties. Oxidative stress is regarded as an important starting factor for neuronal cell loss and necrosis, is one of the causes of Parkinson’s disease (PD), and is considered to be the cause of adverse reactions caused by the current PD commonly used treatment drug levodopa (l-DA). Supplementation with antioxidants early in PD can effectively prevent neurodegeneration and inhibit apoptosis in dopaminergic neurons. At present, the effect of fucoxanthin in improving the adverse effects triggered by long-term l-DA administration in PD patients is unclear. In the present study, we found that fucoxanthin can reduce cytotoxicity and suppress the high concentration of l-DA (200 μM)-mediated cell apoptosis in the 6-OHDA-induced PC12 cells through improving the reduction in mitochondrial membrane potential, suppressing ROS over-expression, and inhibiting active of ERK/JNK-c-Jun system and expression of caspase-3 protein. These results were demonstrated by PD mice with long-term administration of l-DA showing enhanced motor ability after intervention with fucoxanthin. Our data indicate that fucoxanthin may prove useful in the treatment of PD patients with long-term l-DA administration.

Atmospheric ammonia causes histopathological lesions, cell cycle blockage, and apoptosis of spleen in chickens

This experiment was conducted to investigate the effect of atmospheric ammonia (NH3) on histological changes, cell cycle distribution, and apoptosis of the spleen in chickens. Two hundred forty chickens were randomly allocated to the control group (without NH3 challenge) and NH3 group (70 ± 5 ppm NH3). The experiment lasted for 8 days. The results showed that NH3 exposure caused decreased relative weight ( P &lt; 0.05), dysplasia of white pulps, obvious ultrastructural lesions, upregulation of G0G1 phase cells, excessive apoptosis, and oxidative stress ( P &lt; 0.05) in the spleen. The mechanisms of cell cycle blockage were closely related to the upregulation of the P53 and P21 genes ( P &lt; 0.05), the downregulation of the cyclin D1 and CDK6 genes ( P &lt; 0.05), and the decrease of proliferating cell nuclear antigen (PCNA) protein ( P &lt; 0.05). The activated apoptosis resulted from the increased gene and protein expression of Bax and caspase-3 ( P &lt; 0.05), and the decreased gene and protein expression of Bcl-2 ( P &lt; 0.05). The results suggested that 70 ± 5 ppm NH3 caused the spleen dysplasia, which was closely related to the cell cycle arrest and mitochondrial apoptotic pathway activation.

Tert-butylhydroquinone prevents neuroinflammation and relieves depression via regulation of NLRP3 signaling in mice

Depression is a common psychiatric disorder, which seriously affects people’s health and quality of life. Current treatments, which mainly focus on neurotransmitter levels, are not effective in many patients. Recent studies have shown that neuroinflammation has certain correlation with the pathogenesis of depression. Tert-butylhydroquinone (TBHQ) is an antioxidant with an anti-inflammatory effect. The present study evaluated the effects of TBHQ on the improvement of depression-like behaviors induced by lipopolysaccharide (LPS) in mice and its possible mechanism. Behavioral test results showed that TBHQ treatment could significantly improve the depression-like behaviors of mice. Western blot results showed that TBHQ treatment inhibited the protein expression of NLRP3, Caspase-1, IL-1β, and IL-18, which induced by LPS. Immunofluorescence staining results showed that TBHQ treatment inhibited the activation of microglia induced by LPS. These results suggested that, by inhibiting LPS-induced neuroinflammation and microglia activation, TBHQ could effectively improve LPS-induced inflammation-related depression-like behavior through modulating the NLRP3 signaling pathway.

Quercetin inhibits cytotoxicity of PC12 cells induced by amyloid-beta 25–35 via stimulating estrogen receptor α, activating ERK1/2, and inhibiting apoptosis

Abstract The accumulation of β-amyloid (Aβ) in the brain plays an important role in the pathogenesis of Alzheimer’s disease (AD). The lack of estrogen is one of the risk factors for AD. Quercetin is a phytoestrogen with a chemical structure similar to that of estrogen. However, the mechanism by which quercetin prevents AD is unclear. PC12 cells were cultured with Aβ25–35 for 24 h. Then the cells were further treated with 17β-estradiol, genistein, and quercetin for another 24 h, respectively. Next, ICI182780 and U0126 were used to study the mechanisms of estrogen-like neuroprotection. Methyl thiazolyl tetrazolium (MTT) assay was performed to detect cell survival. The protein expression was analyzed by immunofluorescence and western blot. The survival of PC12 cells induced by Aβ25–35 was increased by quercetin. The levels of estrogen receptor α (ERα) and p-extracellular signal-regulated kinase (ERK)1/2 were improved by quercetin, but not those of ERβ. On the contrary, Bcl-2/Bax was increased and the expression of Caspase-3 was decreased. When the cell was pretreated with ICI182780, the p-ERK1/2 and Bcl-2/Bax ratio was decreased, but Caspase-3 expression was increased. In addition, pretreatment with U0126 would reduce Bcl-2/Bax ratio and increase Caspase-3 protein expression. Conclusively, quercetin plays a neuroprotective role through the ER pathway and the mitogen-activated protein kinase (MAPK) pathway. The MAPK signaling pathways could also be activated by quercetin via the mediation of ERα.

Membrane-Free Stem Cells and Pyridoxal 5'-Phosphate Synergistically Enhance Cognitive Function in Alzheimer's Disease Mouse Model.

Accumulation of amyloid beta (Aβ) is a major pathological hallmark of Alzheimer’s disease (AD). In this study, we evaluated the protective effect of membrane-free stem cell extract (MFSCE), which is a component of adipose-tissue-derived stem cells, on cognitive impairment in Aβ25–35-injected AD mice. The ICR mice were i.c.v. injected with Aβ25–35 and then treated with MFSCE for 14 days (i.p.). The Aβ25–35-injected mice showed deficits in spatial and object perception abilities, whereas treatment with MFSCE inhibited Aβ25–35-induced learning and memory impairment in the T-maze, novel object recognition, and Morris water maze tests. Moreover, Aβ25–35-induced lipid peroxidation and nitric oxide overproduction were attenuated by treatment with MFSCE. These antioxidant effects of MFSCE were related to the inhibition of the apoptotic signaling pathway. In particular, the combination treatment of MFSCE and pyridoxal 5′-phosphate (PLP) showed greater suppression of Bax and cleaved caspase-3 protein expression compared to the MFSCE- or PLP-only treatment. Furthermore, the MFSCE and PLP combination significantly downregulated the amyloidogenic-pathway-related protein expressions, such as amyloid precursor protein, presenilin 1, and presenilin 2. Therefore, the MFSCE and PLP combination may synergistically prevent Aβ25–35-induced neuronal apoptosis and amyloidogenesis, which contributes to cognitive improvement and has potential therapeutic implications for AD patients.

Effects and mechanism of hydrogen peroxide pretreatment with low molarity on oxidative stress induced apoptosis of mouse bone marrow mesenchymal stem cells

Effects and mechanism of hydrogen peroxide pretreatment with low molarity on oxidative stress induced apoptosis of mouse bone marrow mesenchymal stem cells

Can metformin modulate the retinal degenerative changes in a rat model of retinitis pigmentosa?

Retinitis pigmentosa (RP) affects over a million people worldwide, characterized by photoreceptor cell death, progressive retinal degeneration, and visual loss. Metformin is demonstrated as a potential therapeutic approach for preventing light-induced retinal degeneration by decreasing apoptosis and oxidative stress. This work aimed to investigate the effect of metformin on the retina of the N-Ethyl-N-nitrosourea (ENU) induced rat model of RP. Eighteen adult male Wistar rats were divided into two groups. Group I: normal vehicle control (N = 6). Group II: ENU-induced photoreceptor degeneration (N = 12) received a single intraperitoneal injection of ENU at a 600 mg/kg dose. Rats in group II were equally divided into two subgroups: IIa: photoreceptor degeneration-induced group and IIb: metformin-treated group (200 mg/kg) for seven days. Specimens from the retina were processed for light and electron microscopy. In ENU treated group, the retina revealed vacuolations and morphological changes in the glia (Müller cells and microglia) and blood capillaries. Increasing caspase-3 (apoptotic marker), iNOS (oxidative stress marker), CD68 (macrophage marker) and glial fibrillary acidic protein (GFAP) expression were detected. In the metformin-treated group, the retinal vacuolations reduced with the morphological improvement in the glia and blood capillaries. Caspase-3, iNOS, CD68, and GFAP expression decreased. Metformin was found to have a neuroprotective effect on the retina in ENU induced rat model of RP

Dietary Restriction May Attenuate the Expression of Cell Death–Related Proteins in Rats with Acute Spinal Cord Injury

There is currently no effective treatment for spinal cord injuries (SCIs). Previous studies have shown that every-other-day fasting (EODF), a dietary restriction method, can reduce SCI size and promote motor function recovery, making it a potential novel treatment. However, the mechanism that underlies the positive impact of EODF on SCI remains unclear. Caspase-dependent apoptosis and necroptosis, which involve receptor-interacting protein kinase (RIPK), drive the loss of nerve cells and restrict motor function recovery after SCI. Dietary restriction has a significant inhibitory effect on Caspase and RIPK expression. This study aimed to investigate whether the EODF diet achieves a neuroprotective effect by inhibiting Caspase-dependent apoptosis and RIPK-dependent necroptosis after SCI. The model rats underwent EODF for 4weeks before SCI or started EODF diet immediately after SCI. Immunoblotting and immunohistochemical analyses were used to assess the impact of the intervention on protein expression. Apoptosis in the spinal cord was detected by TdT-mediated dUTP nick-end labeling. Immunoblotting analysis results revealed that the levels of both RIPK1 and RIPK3 proteins in the injury zone were reduced at 6, 12, and 24hours and at 3 and 7days after SCI, respectively. Immunohistochemistry results showed that EODF reduced the expression of Caspase-3 and Bax proteins, while prophylactic EODF decreased the rate of apoptosis detected by TdT-mediated dUTP nick-end labeling within 3days after SCI. These findings indicate that the mechanism by which EODF exerts neuroprotective effects may be related to the simultaneous inhibition of apoptosis and necroptosis in SCI.

Picroside II alleviates DSS-induced ulcerative colitis by suppressing the production of NLRP3 inflammasomes through NF-κB signaling pathway

Context Ulcerative colitis (UC) is a common acute or chronic intestinal disease with an imbalance of inflammation. Picroside II (P-II) exerts a protective role in various inflammation-related diseases. However, the effect of P-II on UC is still unclear. Objective To explore the effect of P-II on UC and its potential mechanism. Materials and methods Human monocytic leukemia cell line THP-1 was treated with phorbol ester (PMA) to differentiate into a macrophage. The differentiated THP-1 cells were hatched with LPS combined with ATP or Nigericin to activate the NLRP3 inflammasome in vitro. The UC model was constructed by injection of DSS into mice. Results The maximum nontoxic concentration of P-II on THP-1 cells was 60 μM. LPS combined with ATP or Nigericin stimulated the production of IL-1β, which was antagonized by P-II treatment. Meanwhile, P-II administration interfered with the aggregation of ASC and the assembly of NLRP3 inflammasomes. Also, P-II treatment reduced the LPS and ATP-induced elevation of the relative protein expression of NLRP3, pro-caspase-1, IL-1β and p-p65/p65, and the concentration of TNF-α and IL-6. Besides, the NF-κB specific inhibitor BAY-117082 notably repressed the LPS together with ATP-enhanced the relative protein expression of NLRP3, caspase-1 and IL-1β. Moreover, in vivo results showed that P-II relieved the DDS-induced UC, as evidenced by the improvement of mice weight, DAI and pathological scores. In addition, P-II treatment notably decreased DDS-promoted expression of NLRP3 inflammasomes and inflammatory factors in vivo. Conclusion P-II alleviated DSS-induced UC by repressing the production of NLRP3 inflammasomes via the NF-κB signaling pathway.

NaAsO2 decreases GSH synthesis by inhibiting GCLC and induces apoptosis through Hela cell mitochondrial damage, mediating the activation of the NF-κB/miR-21 signaling pathway

BackgroundCervical cancer is the fourth most common cancer in women worldwide, and arsenic has a certain effect in solid tumor chemotherapy. As the rate-limiting enzyme subunit of GSH synthesis, GCLC may be an important target for arsenic to induce apoptosis through mitochondrial apoptosis pathway to exert anti-tumor effect. NF-κB plays an important role in the occurrence and development of cervical cancer and can regulate the expression of GCLC. miR-21 is a potential biomarker of cervical cancer, which can induce apoptosis through ROS regulated the mitochondrial pathway of cells. However, the role of miR-21 in the mitochondrial pathway of cervical cancer cells induced by NaAsO2 through NF-κB/GCLC and GSH synthesis regulated oxidative stress is rarely reported. Therefore, the purpose of this study was to investigate whether NaAsO2 might induce mitochondrial damage and apoptosis of cervical cancer cells through NF-κB/ miR-21 /GCLC induced oxidative stress, and play the anti-tumor role of arsenic as a potential drug for the treatment of cervical cancer. MethodsHela cells were treated with different concentrations of NaAsO2, D, L-Buthionine-(SR)-sulfoximine (BSO), IκBα inhibitor (BAY 11-7082) and miR-21 Inhibitor. CCK-8 assay, Western Blot, qRT PCR, immunofluorescence, transmission electron microscopy, mitochondrial Membrane Potential Assay Kit with JC-1,2′,7′-Dichlorofluorescin diacetate fluorescent probe and Annexin V-FITC were used to measure cell activity, GSH and ROS, mitochondrial morphology and membrane potential (ΔΨm), protein and mRNA expression of GCLC, GCLM, p65, IκBα, p-P65, p-I κBα, Bcl-2, BAX, Caspase3, cleaved-caspase3 and miR-21. ResultsCompared with the control group, with the gradual increasing dose of NaAsO2, cell viability was considerable reduced, and increased rate of apoptosis, intracellular GSH level was decreased significantly, ROS was increased, mitochondrial structure was damaged, mitochondrial membrane potential ΔΨm and Bcl2/BAX lowered, the expression of Caspase3 and cleaved-caspase3 were significantly increased, resulting in mitochondrial apoptosis. When Hela cells were treated with 15, 20, and 25 μmol/L NaAsO2, the mRNA and protein levels of GCLC and GCLM were reduced, the expression of p65 in the nucleus was increased, the expression of p-p65/p65, p-IκBα/IκBα and miR-21 were significantly increased. When BSO increased the inhibitory effect of NaAsO2 on GCLC, Compared with NaAsO2 group, the ΔΨm and protein of Bcl-2/BAX, caspase3 and cleaved-capsase3 were increased. When BAY 11–7082 combined with NaAsO2 co-treated, compared with the NaAsO2 group, the protein and mRNA expression of GCLC was increased, NaAsO2-increased expression level of miR-21 was suppressed, and the ΔΨm and cell viability were higher. In addition, compared with the combination of NaAsO2 and miR-21NC, the protein expression of GCLC was increased, the ΔΨm and cell viability reduction were alleviated by miR-21 Inhibitor combined with NaAsO2. ConclusionNaAsO2 may lead to ROS accumulation in Hela cells and trigger mitochondrial apoptosis. The mechanism may be related to the activation of NF-κB signaling pathway and the promotion of miR-21 expression which leads to the inhibition of GCLC expression and the significant decrease of intracellular reductive GSH synthesis.

Fenofibrate Exerts Anticancer Effects on Human Cervical Cancer HeLa Cells via Caspase-Dependent Apoptosis and Cell Cycle Arrest

Objective: In the present study, we attempted to identify the effects of fenofibrate on human cervical cancer cells. Methods: The cytotoxicity of fenofibrate in cervical cancer cells was determined by Cell Counting Kit-8. Immunoblotting assay was used to determine the protein expression of caspase-3, poly ADP-ribose polymerase cleavage, B-cell lymphoma 2 family protein expression, microtubule-associated protein 1A/1B-light chain 3 (LC3), as well as cyclins and cyclin-dependent kinases. Immunofluorescence imaging was used to determine the expression of cleaved caspase-3 and LC3. Flow cytometry was used to determine cell cycle and apoptosis. Results: We first showed that fenofibrate treatment reduced cell viability in HeLa cervical cancer cells in a dose-dependent manner at 24 h and 48 h. Importantly, fenofibrate-induced cell death was mediated through cell cycle arrest in the G0–G1 phase and caspase-dependent apoptosis. Moreover, fenofibrate also induced autophagy activation in a dose-dependent manner and pharmacological inhibition of autophagy led to increase of sub-G1 phase and caspase-dependent cell death in HeLa cells. Conclusion: In conclusion, these data demonstrated that fenofibrate initially induced cell cycle arrest, followed by caspase-3-dependent cell death in cervical cancer HeLa cells. However, fenofibrate also induced autophagy activation, which is closely related to the survival of diverse cancer cells, thus reducing the anticancer effects of fenofibrate. Therefore, the combination of an autophagy inhibitor and fenofibrate might have the potential to become a new therapeutic strategy for cervical cancer.

Molybdenum and cadmium co-exposure induces endoplasmic reticulum stress-mediated apoptosis by Th1 polarization in Shaoxing duck (Anas platyrhyncha) spleens

Excessive molybdenum (Mo) and cadmium (Cd) are deleterious to animals, but immunotoxicity co-induced by Mo and Cd remains unclear. To ascertain the confederate impacts of Mo and Cd on endoplasmic reticulum (ER) stress-mediated apoptosis by Helper T (Th) cells 1 polarization in the spleen of ducks, we randomly allocated forty 8-day-old Shaoxing ducks (Anas platyrhyncha) into 4 groups and reared them with having different doses of Mo and/or Cd basic diet. At the 16th week of the experiment, serum and spleen tissues were extracted. Data confirmed that Mo and/or Cd strikingly promoted their levels in spleen, caused histological abnormality and trace elements imbalance, and disrupted Th1/Th2 balance to divert toward Th1, then triggered ER stress by increasing three branches PERK/eIF2α/CHOP, IRE1/Caspase-12 and TRAF2/JNK signaling pathways-related genes mRNA and proteins levels, which stimulated apoptosis by elevating Bak-1, Bax, Caspase-9, Caspase-3 mRNA expression, and cleaved-Caspase-9/Caspase-9, cleaved-Caspase-3/Caspase-3 proteins expression as well as apoptosis rate, and decreasing Bcl-xL, Bcl-2 mRNA expression and Bcl-2/Bax ratio. Besides, the variation in combined group was most evident. Briefly, the study indicates that Mo and/or Cd exposure trigger ER stress-induced apoptosis via Th1 polarization in duck spleens, and its mechanism is somehow closely linked with the deposition of Cd and Mo, which may aggravate toxic damage to spleen.

Effect of high-intensity interval training and coenzyme Q10 supplementation on cardiac apoptosis in obese male rats.

Obesity is associated with several metabolic disorders and myocardial apoptosis. The aim of this study was to determine the effect of high-intensity interval training (HIIT) and coenzyme Q10 (CoQ10) supplementation on myocardial apoptotic proteins in obese male rats. Forty-eight male Wistar rats were randomly assigned to 6 groups including non-obese control (NOC), baseline obese control (BOC), HIIT, CoQ10, HIIT + CoQ10, and control. NOC and BOC groups were sacrificed at the beginning of the study. Exercise groups were subjected to a HIIT program over 12 weeks. Rats in CoQ10 group were gavage-fed with 500 mg/kg-1 CoQ10 supplement throughout the study. The heart muscle was removed 48 hours after the last training session. The B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), cytochrome c, BH3-interacting domain death agonist (Bid), caspase-8, and caspase-3 protein expressions were analyzed using western blotting. Data were analyzed using independent t-test and two-way analysis of variance (ANOVA). The Bax, Bid, cytochrome c, caspase-8, caspase-3 proteins, and Bax/Bcl-2 were significantly higher in BOC group compared to NOC (P = 0.025, P = 0.0001, P = 0.013, P = 0.017, P = 0.010, P = 0.180, respectively). Moreover, Bcl-2 protein was lower in BOC compared to NOC group (P = 0.025). HIIT program decreased Bax/Bcl-2 ratio (P = 0.012), Bid (P = 0.0001), cytochrome c (P = 0.003), caspase-8 (P = 0.006), and caspase-3 (P = 0.002) proteins and increased Bcl-2 (P = 0.001) proteins in heart muscle. CoQ10 supplementation significantly increased Bcl-2 protein content (P = 0.001). HIIT exerts an anti-apoptotic effect in heart muscle of obese rats. Although the administration of CoQ10 increased Bcl-2 anti-apoptotic protein, it did not show a potent synergistic effect along with HIIT to reduce obesity-related myocardial apoptosis.

Notoginsenoside R1 relieves the myocardial infarction via activating the JAK2/STAT3 signaling pathway in vivo and in vitro

ABSTRACT Myocardial infarction (MI), caused by continuous ischemia and hypoxia of the coronary artery, is one of the major causes of human mortality. This study aimed to investigate the role of notoginsenoside R1 (NGR1) in MI therapy. In vitro and in vivo models of MI were established by hypoxia/reoxygenation (H/R)-treatment of H9C2 cells and through the ligation of the left anterior descending coronary artery of rats, respectively. CCK-8 and EdU assays were performed to measure cell viability and proliferation, respectively. Flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining were performed to determine the apoptotic rate of cells. Western blot was used to determine protein expression. The MI area was analyzed by 2,3,5-triphenyltetrazolium chloride (TTC) staining. NGR1 promoted viability and proliferation, and inhibited the apoptotic rate of H/R-treated H9C2 cells. In addition, NGR1 downregulated the protein expression of caspase-3 and Bax, and upregulated Bcl-2 expression in H/R-treated H9C2 cells. The JAK2/STAT3 signaling pathway was activated following NGR1 treatment in vivo and in vitro, and inhibition of the JAK2/STAT3 signaling pathway reversed the effects of NGR1 on H/R-treated H9C2 cells. Finally, NGR1 reduced the area of MI. NGR1 relieved MI in vivo and in vitro by activating the JAK2/STAT3 signaling pathway.

Fluvastatin alleviates doxorubicin-induced cardiac and renal toxicity in rats via regulation of oxidative stress, inflammation, and apoptosis associated genes expressions

Doxorubicin (DOXO) is a cytostatic agent used in the chemotherapy protocol of several cancers for more than 40 years, but usage of this drug in cancer treatment has been limited due to severe renal and cardiac tissue toxicities that may result in death in patients. Fluvastatin (FV) is a fully synthetic hydroxymethyl glutaryl coenzyme A (HMG-CoA) reductase inhibitor used as a cholesterol-lowering agent in patients with hypercholesterolemia. Previous studies revealed that FV also exhibits antioxidant, anti-inflammatory, and antitumor activity. Additionally, our previous study indicated that FV exerts a prophylactic effect on DOXO-induced testicular toxicity by preventing lipid peroxidation, supporting the antioxidant system, and regulating the blood-testis barrier-associated genes expression. Herein, we purposed to evaluate the possible therapeutic and the protective effects of FV on the DOXO-induced cardiac and renal toxicitiy model by histochemical, immunohistochemical, biochemical, and real-time polymerase chain reaction (real-time PCR) analyses. Results point out protective use of FV exerts a beneficial effect by repressing lipid peroxidation and by regulating the inducible nitric oxide synthase (iNOS), nitric oxide synthase endothelial (eNOS), nuclear factor kappa-B (NF-κB), and Caspase-3 (Casp3) protein and mRNA expressions, which play an important role in mediating DOXO-induced renal and cardiac toxicity mechanisms. In conclusion, FV may be a candidate agent for the prevention of renal and cardiac toxicities in cancer patients receiving DOXO chemotherapy.

Dexmedetomidine Alleviates Lung Oxidative Stress Injury Induced by Ischemia-Reperfusion in Diabetic Rats via the Nrf2-Sulfiredoxin1 Pathway.

Oxidative stress injury (OSI) is an important pathological process in lung ischemia-reperfusion injury (LIRI), and diabetes mellitus (DM) can exacerbate this injury. Dexmedetomidine protects against LIRI by reducing OSI. However, the effect of dexmedetomidine on LIRI under diabetic conditions remains unclear. Therefore, this study is aimed at exploring the effects and mechanisms of dexmedetomidine on OSI induced by LIRI in diabetic rats. Rats were randomly divided into control+sham (CS), DM+sham (DS), control+ischemia-reperfusion (CIR), DM+ischemia-reperfusion (DIR), and DM+ischemia-reperfusion+dexmedetomidine (DIRD) groups (n = 6). In the CS and DS groups, the nondiabetic and diabetic rats underwent thoracotomy only without LIRI. In the CIR, DIR, and DIRD groups, LIRI was induced through left hilum occlusion for 60 min, followed by reperfusion for 120 min in nondiabetic and diabetic rats, and rats in the DIRD group were administered dexmedetomidine (3, 5, and 10 μg/kg). Compared with those in the CS group, the OSI, lung compliance, apoptosis, and oxygenation indices deteriorated in the DS group (P < 0.05), and these indices were further aggravated in the CIR and DIR groups (P < 0.05), being the worst in the DIR group (P < 0.05). Compared to those of the DIR group, the OSI, lung compliance (15.8 ± 2.4 vs. 11.6 ± 1.7 ml/kg), apoptosis (22.5 ± 2.6 vs. 51.8 ± 5.7), oxygenation (381 ± 58 vs. 308 ± 78 mmHg), and caspase-3 and caspase-9 protein expression indices were attenuated, and Nrf2 and sulfiredoxin1 protein expression was increased in the DIRD group (P < 0.05). And the lung injury, oxygenation, OSI, and Nrf2 and sulfiredoxin1 protein expression changed in a concentration-dependent manner. In conclusion, dexmedetomidine alleviated lung OSI and improved lung function in a diabetic rat LIRI model through the Nrf2-sulfiredoxin1 pathway.

Sevoflurane preconditioning prevents acute renal injury caused by ischemia-reperfusion in mice via activation of the Nrf2 signaling pathway.

Oxidative stress, caused by renal ischemia reperfusion (IR)/hypoperfusion, is one of the main causes of acute kidney injury (AKI). Previous studies have demonstrated that sevoflurane (SEV) protects organs from the damage caused by oxidative stress. In the present study, mice were randomly assigned to a sham operation group (Sham), IR-vehicle group (IR+ vehicle), IR + SEV low-dose preconditioning group and an IR + SEV high-dose preconditioning group. The effect of SEV on nuclear factor E2-related factor 2 (Nrf2), a key regulatory protein of the endogenous antioxidant defense system and, consequently oxidative stress, inflammation and apoptosis-related factors, were all quantified using commercial kits or by western blotting. SEV preconditioning was demonstrated to ameliorate kidney injury as a result of decreased blood urine nitrogen and serum creatinine levels, activated Nrf2 expression in the kidney and decreased oxidative stress and inflammatory index levels an AKI mouse model. SEV preconditioning also protected injured kidney via the downregulation of caspase-3 protein expression levels. In addition, using the Nrf2 inhibitor, Brusatol, significantly abolished the SEV preconditioning renal protective effect. Using an in vitro HK-2 cell model of hypoxia/reoxygenation, it was also demonstrated that Nrf2 pathway activation was necessary for SEV to exert its beneficial effect for tubular cell injury caused by hypoxia/reoxygenation. These results indicated that SEV may protect against renal injury caused by IR via Nrf2 upregulation.