Caspase-3 mRNA Expression Research Articles

Temporal patterns and distribution of pyroptosis-related molecules and effects of human mesenchymal stem cells on pyroptosis following cerebral ischemia/reperfusion in rats

ObjectivesPyroptosis is a new type of programmed cell death that has a strong proinflammatory effect. The present study investigated the dynamic changes of pyroptosis-related molecules and the effect of mesenchymal stem cells (MSCs) on pyroptosis following cerebral ischemia/reperfusion (I/R). Materials and MethodsThe temporal pattern and cellular distribution of caspase-1, Gasdermin D and E (GSDMD and GSDME) in the peri-infarct area, and the effect of human MSCs on GSDMD, IL-1β, IL-18, Lactate dehydrogenase (LDH) and neurological function were studied in a rat model of transient focal cerebral ischemia. ResultsThe expression of caspase-1 mRNA increased with time, with a protein level of pro-caspase-1 comparable to its mRNA level, while the level of cleaved-caspase-1 protein peaked at 48 h following I/R. Increased levels of GSDMD mRNA and protein were also observed, with a peak level at 24 h. There were no significant changes in GSDME mRNA or protein expression after I/R. In regards to changes in the number of cells expressing GSDMD after I/R, that for neurons was more significant than those for microglia and astrocytes. The modified neurological severity score discrepancy and the expression of GSDMD showed no significant differences within 24 h following I/R between the MSC- and NS-treated groups, but MSCs treatment promoted the secretion of IL-1β, IL-18 and LDH. ConclusionsIn the early stage of cerebral infarction in rats, there were dynamic changes in pyroptosis-related molecules (caspase-1 and GSDMD), but MSCs showed no effect on the levels of GSDMD or neurological function.

Baicalin Attenuates H2O2-Induced Oxidative Stress by Regulating the AMPK/Nrf2 Signaling Pathway in IPEC-J2 Cells.

Oxidative stress can adversely affect the health status of the body, more specifically by causing intestinal damage by disrupting the permeability of the intestinal barrier. This is closely related to intestinal epithelial cell apoptosis caused by the mass production of reactive oxygen species (ROS). Baicalin (Bai) is a major active ingredient in Chinese traditional herbal medicine that has antioxidant, anti-inflammatory, and anti-cancer properties. The purpose of this study was to explore the underlying mechanisms by which Bai protects against hydrogen peroxide (H2O2)-induced intestinal injury in vitro. Our results indicated that H2O2 treatment caused injury to IPEC-J2 cells, resulting in their apoptosis. However, Bai treatment attenuated H2O2-induced IPEC-J2 cell damage by up-regulating the mRNA and protein expression of ZO-1, Occludin, and Claudin1. Besides, Bai treatment prevented H2O2-induced ROS and MDA production and increased the activities of antioxidant enzymes (SOD, CAT, and GSH-PX). Moreover, Bai treatment also attenuated H2O2-induced apoptosis in IPEC-J2 cells by down-regulating the mRNA expression of Caspase-3 and Caspase-9 and up-regulating the mRNA expression of FAS and Bax, which are involved in the inhibition of mitochondrial pathways. The expression of Nrf2 increased after treatment with H2O2, and Bai can alleviate this phenomenon. Meanwhile, Bai down-regulated the ratio of phosphorylated AMPK to unphosphorylated AMPK, which is indicative of the mRNA abundance of antioxidant-related genes. In addition, knockdown of AMPK by short-hairpin RNA (shRNA) significantly reduced the protein levels of AMPK and Nrf2, increased the percentage of apoptotic cells, and abrogated Bai-mediated protection against oxidative stress. Collectively, our results indicated that Bai attenuated H2O2-induced cell injury and apoptosis in IPEC-J2 cells through improving the antioxidant capacity through the inhibition of the oxidative stress-mediated AMPK/Nrf2 signaling pathway.

Design, Synthesis and Biological Evaluation of Multi-Target Anti-Cancer Agent PYR26.

This study investigates the synthesis of a new compound, PYR26, and the multi-target mechanism of PYR26 inhibiting the proliferation of HepG2 human hepatocellular carcinoma cells. PYR26 significantly inhibits the growth of HepG2 cells (p < 0.0001) and this inhibition has a concentration effect. There was no significant change in ROS release from HepG2 cells after PYR26 treatment. The mRNA expressions of CDK4, c-Met and Bak genes in HepG2 cells were significantly inhibited (p < 0.05), while mRNA expression of pro-apoptotic factors such as caspase-3 and Cyt c was significantly increased (p < 0.01). The expression of PI3K, CDK4 and pERK proteins decreased. The expression level of caspase-3 protein was increased. PI3K is a kind of intracellular phosphatidylinositol kinase. PI3K signaling pathway is involved in signal transduction of a variety of growth factors, cytokines and extracellular matrix and plays an important role in preventing cell apoptosis, promoting cell survival and influencing cell glucose metabolism. CDK4 is a catalytic subunit of the protein kinase complex and is important for G1 phase progression of the cell cycle. PERK refers to phosphorylated activated ERK, which is translocated from cytoplasm to the nucleus after activation, and then participates in various biological reactions such as cell proliferation and differentiation, cell morphology maintenance, cytoskeleton construction, cell apoptosis and cell canceration. Compared with the model group and the positive control group, the tumor volume of the nude mice in the low-concentration PYR26 group, the medium-concentration group and the high-concentration group was smaller, and the organ volume was smaller than that in the model group and the positive control group. The tumor inhibition rates of low-concentration group PYR26, medium-concentration group and high-concentration group reached 50.46%, 80.66% and 74.59%, respectively. The results showed that PYR26 inhibited the proliferation of HepG2 cells and induced apoptosis of HepG2 cells by down-regulating c-Met, CDK4 and Bak, up-regulating the mRNA expression of caspase-3 and Cyt c genes, down-regulating PI3K, pERK and CDK4 proteins and up-regulating the protein level of caspase-3. In a certain range, with the increase in PYR26 concentration, the tumor growth was slower and the tumor volume was smaller. Preliminary results showed that PYR26 also had an inhibitory effect on the tumors of Hepa1-6 tumor-bearing mice. These results suggest that PYR26 has an inhibitory effect on the growth of liver cancer cells, therefore it has potential to be developed into a new anti-liver cancer drug.

Ameliorative effect of cheqianzi decoction on hyperuricemia and kidney injury and underlying mechanism in rats

Cheqianzi Decoction (CQD) is a Traditional Chinese Medicine (TCM) formula comprising four herbs and is recorded in the Ancient Materia Medica “Shengji Zonglu”. Individually, these four herbs have been shown to reduce uric acid (UA) levels, to treat hyperuricemia (HUA), and alleviate kidney damage. However, the therapeutic efficacy of the CQD and related mechanism are not yet clear.In this study, high performance liquid chromatography (HPLC) analysis confirmed that the contents of the chemical components of the four herbal medicines were in accordance with the provisions of the Chinese Pharmacopoeia. A total of 99 potential targets were identified in the network pharmacology analysis of CQD, indicating its involvement in the regulation of inflammatory and apoptotic signaling pathways, and potential value for treating HUA and alleviating kidney injury. In vivo pharmacodynamic studies showed that compared with the Model group, significantly decreased levels of serum uric acid (SUA), serum creatinine (SCr), blood urea nitrogen (BUN) (all P < 0.05), and inflammatory factors (P < 0.01) were detected in the CQD group. Quantitative real-time PCR and Western blot analyses showed that compared with the Model group, adenosine triphosphate (ATP)-binding cassette efflux transporter G2 (ABCG2) expression in the CQD group was significantly upregulated (P < 0.01) at both the mRNA and protein levels, while mRNA expression of Caspase3 and NOD-like receptor family member 3 (NLRP3) (P < 0.05) and protein expression of NLRP3 (P < 0.01) were significantly downregulated.In conclusion, CQD promotes UA excretion by activating ABCG2, and induces inflammasome NLRP3-mediated reduction in inflammatory and apoptotic factors to achieve renal protection. Thus, our findings indicate the therapeutic potential of CQD in HUA with kidney injury.

Investigation of the relationship between NFKB1 polymorphisms and telomere length and apoptosis in patients with type-2 diabetes

Purpose: Type 2 diabetes mellitus (T2DM) is a heterogeneous, chronic, and metabolic disease that affects a significant proportion of the global population. This study aimed to evaluate the effect of NFKB1 -94 ATTG ins/del polymorphisms on the expression of apoptosis genes and telomere length (TL) in patients with T2DM compared with healthy individuals.&#x0D; Materials and Methods: Sixty-nine T2DM patients and sixty healthy people were enrolled in the study. DNA and RNA were isolated from the blood samples. NFKB1 genotypes were identified by Sanger sequencing. For TL analyses and to investigate the expression of the caspase-3, caspase-9, bax, and bcl2 genes, RT-PCR was utilized.&#x0D; Results: There was a significant difference between the NFKB1 -94 ins/del genotype patients and the control group (OR:0.4792 (0.2345-1.011)). However, the distribution of other genotype/alleles (ins/ins and del/del) showed no difference between T2DM and control groups. The allelic frequency of NFKB1 -94 ins/del was 0.455/0.235 for the T2DM group and 0.435/0.165 for the control group. An increase in the mRNA expression of caspase-3, caspase-9 and Bax genes was observed in the T2DM group compared with the healthy group, while a decrease in the Bcl2 gene was found in the T2DM group. TL in T2DM patients was shorter than in healthy individuals. &#x0D; Conclusion: NFKB1 -94 ins/del polymorphisms show significant differences in T2DM patients. We observed that apoptosis was activated and TL was shortened in patients with T2DM. However, no relationship between NFKB1 polymorphisms and apoptosis and TL could not be determined.

The ethyl acetate extract of Wenxia Changfu Formula inhibits the carcinogenesis of lung adenocarcinoma by regulating PI3K-AKT signaling pathway

Lung adenocarcinoma is the most common type of lung cancer. With a rise in new cases worldwide each year, early diagnosis and treatment are very important. Network pharmacology provides the effective way to evaluate poly-pharmacological effects and anticancer molecular mechanisms of drugs. The aim of the present study was to explore the anti-tumor mechanism of ethyl acetate extract of Wenxia Changfu Formula (WFEA) in lung adenocarcinoma by using analytical chemistry, network pharmacology and molecular biology. A total of 193 compounds were identified from WFEA, mainly including esters, phenols, ketones and alkaloids. Totally, 374 targets were regarded as potential targets of WFEA against lung adenocarcinoma. Interestingly, PI3K-AKT was found to be one of the significantly enriched signaling pathways of targets of WFEA against lung adenocarcinoma. AKT1, MMP3, CASP3 and BCL2 had strong binding effect with compound molecules of WFEA. Some combinations with the best docking binding were identified, including quercetin/oleanolic_acid/emodin/aloe_emodin/catechin-AKT1 and quercetin-MMP3. In lung adenocarcinoma cells, the WFEA inhibited the proliferation, migration and invasion, and promoted the apoptosis. Moreover, the WFEA inhibited the mRNA expression of MMP3 and Bcl-2 and promoted the mRNA expression of Caspase3. In addition, WFEA inhibited the protein phosphorylation of AKT and PI3K. The WFEA had a significant inhibitory effect on lung adenocarcinoma cells, which could inhibit cell proliferation, invasion and metastasis, and induce cell apoptosis. The mechanism of action of WFEA may be involved in the regulation of the PI3K-AKT signaling pathway in the lung adenocarcinoma.

Semaphorin 4C Plays a Key Role in Colorectal Cancer Cells Development

Objective: Purpose of this work was to discuss effects and mechanisms of Sema 4C in colon cancer development. Results: Sema4C were significantly upregulated in cancer tissues (P &lt;0.001). Following transfection, the expression levels of Sema4C mRNA were significantly downregulated in the si-Sema4C groups compared with those in the si-negative control groups of both HT-29 and SW620 cells (both P &lt;0.001). The apoptotic rate was significantly increased, while the invasive and wound healing rates were significantly suppressed in the si-Sema4C groups of HT-29 and SW620 cells (both P &lt; 0.001). The results of the RT-qPCR and western blotting analyses revealed that PI3K, AKT, MMP-2 and MMP-9 mRNA and protein expression levels, respectively, were significantly downregulated, while caspase-3 mRNA and protein expression levels were significantly upregulated in the si-Sema4C groups of HT-29 and SW620 cells. Conclusion: The findings of the present study demonstrated that the knockdown of Sema4C expression suppressed CRC cell biological activities by regulating the PI3K/AKT signaling pathway. Therefore, Sema4C may act as an oncogene in CRC.

Effect of pulsed electromagnetic fields on mesenchymal stem cell-derived exosomes in inhibiting chondrocyte apoptosis

The study aims to explore the effect of mesenchymal stem cells-derived exosomes (MSCs-Exo) on staurosporine (STS)-induced chondrocyte apoptosis before and after exposure to pulsed electromagnetic field (PEMF) at different frequencies. The AMSCs were extracted from the epididymal fat of healthy rats before and after exposure to the PEMF at 1 mT amplitude and a frequency of 15, 45, and 75 Hz, respectively, in an incubator. MSCs-Exo was extracted and identified. Exosomes were labeled with DiO fluorescent dye, and then co-cultured with STS-induced chondrocytes for 24 h. Cellular uptake of MSC-Exo, apoptosis, and the protein and mRNA expression of aggrecan, caspase-3 and collagenⅡA in chondrocytes were observed. The study demonstrated that the exposure of 75 Hz PEMF was superior to 15 and 45 Hz PEMF in enhancing the effect of exosomes in alleviating chondrocyte apoptosis and promoting cell matrix synthesis. This study lays a foundation for the regulatory mechanism of PEMF stimulation on MSCs-Exo in inhibiting chondrocyte apoptosis, and opens up a new direction for the prevention and treatment of osteoarthritis.

Plantamajoside alleviates hypoxia-reoxygenation injury through integrin-linked kinase/c-Src/Akt and the mitochondrial apoptosis signaling pathways in H9c2 myocardial cells

Myocardial ischemia–reperfusion injury(MIRI) is one of the common complications after myocardial infarction surgery, Oxidative stress is among the main mechanisms of myocardial ischemia–reperfusion injury. Plantamajoside (PMS), the main effective ingredient in the genus Plantain, has been reported to possess an antioxidation, anti-inflammatory and anti-apoptosis role. However, whether PMS can attenuate myocardial ischemia–reperfusion injury is not yet known. Herein, we explored the effects of PMS on hypoxia-reoxygenation (H/R) injury in H9c2 cardiomyocytes and the underling molecular mechanisms of the treatment. Network pharmacological analysis screened the top 31 key genes in the treatment of MIRI disease treated with PMS, and the result of molecular docking further illustrated the roles that the PMS play in the treatment of MIRI through its interference with integrin-linked kinase (ILK) target protein. PMS was not cytotoxic in the concentration range of 5–40 μM and increased cell survival after H/R injury in a concentration-dependent manner without affecting proliferation or growth. PMS significantly reduced the levels of lactate dehydrogenase, malonic dialdehyde, reactive oxygen species and cell apoptosis, and increased soperoxide dismutase activity compared with those of the H/R injury group. PMS promoted the protein and mRNA expression of ILK and Bcl-2, the protein expression of p-Akt, and reduced the protein and mRNA expression of Bax, Caspase-3, and Cytochrome c, the protein expression of p–c-Src. PMS has protective effects against H/R injury in H9c2 cells, and its protective mechanism may be related to reactive oxygen species clearance, activation of the ILK/c-Src/Akt pathway and inhibition of the mitochondrial apoptosis.

Mechanistic insight into the protective effects of fisetin against arsenic-induced reproductive toxicity in male rats

Arsenic is one of the most hazardous environmental contaminants, which adversely affects the dynamics of male reproductive system. Fisetin (FIS) is a bioactive flavonoid, which is known to exert strong antioxidative effects. Therefore, the current research was planned to evaluate the alleviative efficacy of FIS against arsenic-induced reproductive damages. Forty-eight male albino rats were divided into 4 groups (n = 12), which were treated as follows: (1) Control, (2) Arsenic-intoxicated group (8 mg kg−1), (3) Arsenic + FIS-treated group (8 mg kg−1 + 10 mg kg−1), and (4) FIS-treated group (10 mgkg−1). After 56 days of treatment, the biochemical, lipidemic, steroidogenic, hormonal, spermatological, apoptotic and histoarchitectural profiles of rats were analyzed. Arsenic intoxication reduced the enzymatic activities of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GSR), in addition to glutathione (GSH) level. Conversely, the levels of thiobarbituric acid reactive substance (TBARS) and reactive oxygen species (ROS) were increased. Moreover, it escalated the level of low-density lipoprotein (LDL), triglycerides and total cholesterol, while declining the level of high-density lipoprotein (HDL). Furthermore, steroidogenic enzymes expressions, 3β-hydroxysteroid dehydrogenase (HSD), 17β-HSD, steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (CYP11A1) and 17α-hydroxylase/17, 20-lyase (CYP17A1), were found to be reduced, which brought down the level of testosterone. Besides, the levels of gonadotropins (LH and FSH) were decreased. Additionally, a decline in sperm mitochondrial membrane potential (MMP), motility, epididymal sperm count and hypo-osmotic swelling (HOS) coil-tailed sperms was observed, whereas the dead sperms and structural damages (head, midpiece and tail) of sperms were escalated. Moreover, arsenic exposure up-regulated the mRNA expressions of apoptotic markers, namely Bax and caspase-3, whereas lowered the expression of anti-apoptotic marker, Bcl-2. In addition, it induced histoarchitectural changes in testes of rats. However, FIS treatment resulted in remarkable improvements in testicular and sperm parameters. Therefore, it was inferred that FIS could serve as a therapeutic candidate against arsenic-generated male reproductive toxicity attributing to its anti-oxidant, anti-lipoperoxidative, anti-apoptotic, and androgenic efficacy.

Characterization of Polysaccharides from the Pericarp of Zanthoxylum bungeanum Maxim by Saccharide Mapping and Their Neuroprotective Effects

The pericarp of Zanthoxylum bungeanum maxim (PZM) is a commonly used spice and herbal medicine in China. In the present study, the structural characteristics of PPZM were investigated by saccharide mapping after enzymatic digestion by using high-performance thin layer chromatography (HPTLC) and polysaccharide analysis by using carbohydrate gel electrophoresis (PACE). The mechanisms of protective effects of PPZM on Aβ25-35-induced oxidative damage were explored in PC12 cells. The results showed that PPZM contained 1,4-α-D-galactosidic, 1,4-α-D-galactosiduronic, and (1→4)-β-D-glucosidic linkages. Pretreatment with PPZM significantly increased the cell viability of Aβ25-35-injured PC12 cells. Flow cytometry and Hoechst/PI staining indicated that PPZM gradually relieved the apoptosis of the Aβ25-25-treated cells. PPZM markedly decreased the ROS level of PC12 cells and suppressed Aβ25-35-induced oxidative stress by increasing the SOD level, and decreasing the level of MDA and LDH. The mRNA expressions of caspase-3 and Bax were significantly downregulated, and Bcl-2 expression was upregulated by treatment with PPZM. PPZM significantly increased the mRNA expression of Nrf2 and HO-1 in Aβ25-35 treated cells. The results indicated that PPZM alleviated apoptosis and oxidative stress induced by Aβ25-25 through the inhibition of mitochondrial dependent apoptosis and activation of Nrf2/HO-1 pathway. PPZM can be used as a potential protective agent against Aβ25-25-induced neurotoxicity.

ADSCs-exo attenuates hepatic ischemia-reperfusion injury after hepatectomy by inhibiting endoplasmic reticulum stress and inflammation.

Hepatic ischemia-reperfusion (I/R) injury commonly occurs during liver surgery. Exosomes from adipose-derived stem cells (ADSCs-exo) induce a hepatoprotective effect during hepatic I/R injury. This study aimed to investigate the possible mechanism by which ADSCs-exo attenuates hepatic I/R injury in rats. Rats were randomly divided into four groups: Sham, I30R + PH, ADSCs, and ADSCs-exo groups. Liver tissues were collected immediately after 24 h of reperfusion for further analyses. The content of inflammatory factors in liver tissue was detected using enzyme-linked immunosorbent assay. The pathological changes in liver tissue were analyzed using HE staining. Transmission electron microscopy was used to visualize the ultrastructural changes of hepatocytes. Real-time quantitative polymerase chain reaction(RT-qPCR) and western blot analysis were used to detect the expression of endoplasmic reticulum stress (ERS)-related genes and proteins. Liver histomorphology and hepatocyte ultrastructure changes improved after ADSCs-exo treatment. Moreover, ADSCs-exo treatment significantly downregulated tumor necrosis factor-α,interleukin-1β(IL-1β), and IL-6 levels while upregulating IL-10 levels. Western blot analysis suggested that the protein expressions of GRP78, p-PERK, p-eIF2α, p-IRE1α, XBP1s, ATF-6, ATF-4, CHOP, p-JNK, cleaved-Caspase-3, cleaved Caspase-9, and cleaved Caspase-12 significantly decreased after ADSCs-exo treatment. RT-qPCR results demonstrated that mRNA expression of GRP78, IRE1α, XBP1, ATF-6, ATF-4, CHOP, JNK, Caspase-3, Caspase-9, and Caspase-12 markedly reduced after ADSCs-exo treatment. In conclusion,ADSCs-exo protects against hepatic I/R injury after hepatectomy by inhibiting ERS and inflammation. Therefore, ADSCs-exo can be considered as a viable option for the treatment of hepatic I/R injury.

Investigation of the nephrotoxicity of 2,6-dichloro-1,4-benzoquinone disinfection by-product in mice through a 28-day toxicity test

In recent years, 2,6-dichloro-1,4-benzoquinone (DCBQ) has become an emerging water disinfection by-product and widely distributed in disinfected water. Although kidney is a potential target of DCBQ, a systematic study of the in vivo nephrotoxicity of DCBQ is rare. In this study, a 28-day oral toxicity test was used to assess the nephrotoxic effects of DCBQ on mice. And the potential mechanisms of nephrotoxicity induced by DCBQ were explored through inflammation, oxidative stress, apoptosis and gut microbiota. The results showed that the kidney indexes of mice were not altered in DCBQ-exposed group in comparison with the control group. The histopathological investigation revealed that DCBQ caused swollen of renal tube, destruction of the renal structure, and infiltration of inflammatory cell in kidney. DCBQ has induced oxidative damage in kidney, as the observation of the increase of the renal superoxide dismutase (SOD) and catalase (CAT) activity. Also, DCBQ has triggered the inflammatory response in kidney through the increased expression of IL-1β, NF-κB and iNOS. Moreover, DCBQ has activated the apoptosis pathway, as indicated by the increased mRNA expression of Caspase-3 and Caspase-9. We eventually found an association between gut microbiota and nephrotoxic variables, demonstrating the importance of gut-kidney axis in DCBQ toxicity. Our results suggested that exposure to DCBQ in disinfected water might be a risk factor for kidney and provided novel insights into the underlying mechanisms of DCBQ–induced kidney injury, contributing to better interpretation of the health impact of the environmentally emerging contaminant DCBQ.

Gold-containing liposomes and glucose-coated gold nanoparticles enhances the radiosensitivity of B16F0 melanoma cells via increasing apoptosis and ROS production

AimTo increase the effectiveness of radiation therapy, metals with high Z number are used as radiosensitizers. In this regard, the effectiveness of various gold nanoparticles as radiosensitizer has been proven. Therefore, this study aimed to evaluate the effects of liposomes containing gold ions (Gold-Lips) and glucose-coated gold nanoparticles (Glu-GNPs) on radiation sensitivity of B16F0 melanoma cells. Main methodsNaked GNPs, Glu-GNPs and Gold-Lips were synthesized and their physicochemical properties were evaluated using DLS. The cytotoxicity and sensitivity of the nanoparticles to radiation were evaluated using MTT and colony formation assay, respectively. Flow cytometry was performed to evaluate the apoptotic effect of nanoparticles on B16F0 cells. The intracellular ROS levels and mRNA expression of Bax, Bcl-2, p53, Caspase-3, and Caspase-7 genes were also evaluated. Finally, caspase 3/7 activity was determined using a luminescence assay kit. Key findingsThe results revealed that GNPs, Glu-GNPs, and Gold-Lips had a desired size and zeta potential. Results from the colony assay showed that the all non-toxic concentrations of Gold-Lips significantly increased cell death in B16F0 cells compared with the Glu-GNPs (p > 0.05). Flow cytometry and Caspase-3/-7 activity confirmed the results of the colony assay and showed that increasing the sensitivity of cells to radiation increases apoptosis. Moreover, we found that Gold-Lips increased the mRNA expression of p53, Bax, and Caspase-3/-7, and decreased the Bcl-2 mRNA expression. SignificanceOverall, both Gold-Lips and Glu-GNPs enhanced the radiosensitivity of B16F0 cells, however, Gold-Lips had better effects, which could make them a promising tools in cancer radiotherapy.

Water Hardness Improves the Antioxidant Response of Zinc-Exposed Goldfish (Carassius auratus).

Zinc (Zn), a heavy metal, is an essential element in fish; however, exposure to high concentrations causes oxidative stress. Water hardness reduces oxidative stress reactions caused by heavy metals. To confirm the effect of water hardness on oxidative stress caused by Zn, goldfish were exposed to various Zn concentrations (1.0, 2.0, and 5.0 mg/L) and water hardness (soft (S), hard (H), and very hard (V)). The activity of superoxide dismutase (SOD) and catalase (CAT) in plasma increased with 1.0, 2.0, and 5.0 mg/L of Zn, and decreased with H and V water hardness. The levels of H2O2 and lipid peroxide (LPO) increased with Zn above 1.0 mg/L and decreased with H and V of water hardness. Caspase-9 mRNA expression in the liver increased after 7 and 14 days of Zn exposure and decreased with H and V water hardness. It was confirmed that DNA damage was less dependent on H and V water hardness. Based on the results of this study, at least 1.0 mg/L Zn causes oxidative stress in goldfish, and a high level of apoptosis occurs when exposed for more than 7 days. It appears that the oxidative stress generated by Zn can be alleviated by water hardness of at least 270 mg/L CaCO3. This study provides information on the relationship between the antioxidant response caused by heavy metals and water hardness in fish.

Syringic Acid Suppressed Proliferation, Invasion, and Migration via Inhibition of Matrix Metalloproteinases Expression on Glioblastoma Cells by Promoting Apoptosis.

Human brain tumor glioblastoma (GBM) is the most hostile malignancy, currently lacking a successful cure and good prognosis. To examine the anticancer effects of syringic acid (SA) on human cancer GBM cells. The different doses of SA were added to GBM cells to study its effect on viability, invasion, relocation, apoptosis, and mRNA and protein levels. Hence, we explored the antiproliferative, anti-invasive, and apoptotic activity of SA on GBM human U-251 cells. MTT assay and live/dead assay revealed the anti-proliferative activity of SA on U-251 glioma cells. Apoptotic activity of SA was shown by DAPI staining, caspase-3, Bax, and Bcl-2 mRNA expressions. The cell cycle regulation was also confirmed by reducing the mRNA expression of cyclinD1, CDK4, and CDK6. Treatment of SA with U-251 cells suppressed MMPs expressions and enhanced TIMPs protein levels. Our findings put forward that SA could prevent GBM cells' invasion and relocation. SA is an ideal neuroprotective agent for controlling brain malignancy.

The Mechanism of Aerobic Exercise Regulating the PI3K/Akt-mTOR Signaling Pathway Intervenes in Hippocampal Neuronal Apoptosis in Vascular Dementia Rats.

The purpose of this paper is to explore the mechanism of aerobic exercise regulating autophagy through the PI3K/Akt-mTOR signaling pathway and its participation in apoptosis, to protect the hippocampal nerves from damage in vascular dementia rats. Thirty-six healthy male SD rats were randomly divided into a sham group, a model group, and a model exercise group. A neurobehavioral assessment was used to determine the memory and exploration abilities of the rats. A TUNEL assay was used to detect hippocampal neuron apoptosis. Immunohistochemical and Western blot analyses were used to analyze LC3Ⅱ and the beclin-1 protein. An RT-PCR detected the differential expression of mRNA. The results of the neurobehavioral tests showed that the platform latency time of the rats with vascular dementia was prolonged. Aerobic exercise significantly shortens the swimming time of rats in platform latency. The TUNEL results showed that the TUNEL-positive cells of the hippocampal neurons in the model group increased; the expression of pro-apoptotic genes caspase-3 and Bax mRNA was up-regulated, and the expression of Bcl-2 mRNA was down-regulated. Aerobic exercise reduced hippocampal neuronal apoptosis, up-regulated Bcl-2 mRNA, and down-regulated caspase-3 and Bax mRNA. The LC3Ⅱ and Beclin-1 proteins, detected by immunohistochemistry and a Western blot analysis, showed that the protein expression in the hippocampi of rats with vascular dementia increased. Aerobic exercise reduced LC3Ⅱ and Beclin-1 protein expression. The results of the RT-PCR showed similar changes. Aerobic exercise could improve the learning and memory abilities of vascular dementia rats, moderately regulate the process of autophagy, reduce the TUNEL-positive cells of hippocampal neurons, repair damaged hippocampal neurons by regulating the autophagy signaling pathway PI3K/Akt-mTOR, and improve hippocampal function.

Preparation and characterization of cetuximab-loaded egg serum albumin nanoparticles and their uses as a drug delivery system against Caco-2 colon cancer cells

To avoid the harmful side effects of cetuximab and improve its therapeutic efficacy, egg serum albumin (ESA) was used as a targeting drug carrier moiety for cancer therapy against Caco-2 colon cancer cells. The simple improved desolvation method was used to synthesize ESA nanoparticles (ESA-NPs) and cetuximab-loaded albumin nanoparticles (CET-ANPs) with glutaraldehyde as a crosslinking agent. The ESA-NPs and CET-ANPs were spherically shaped, and their sizes and surface potentials were 100 and − 24 nm and 170 and − 20 nm, respectively, as determined using transmission electron microscopy (TEM) and a Zeta potential analyzer. The specific functional groups of the prepared nanoparticles were revealed by FTIR analysis. In the MTT (3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) assay, CET-ANPs exerted the highest antitumor activity after 24 h followed by CET, ESA-NPs, and pure ESA. Combination of CET + ESA-NPs at different IC50 concentrations at ratios of 1:1, 1:2, 2:1, 1:4, 4:1, 1:9, or 9:1 showed significant synergistic effects with a combination index (CI) > 1. Furthermore, the CET either loaded with ESA-NPs or administered in combination (CET + ESA NPs) caused significant apoptotic damage, as well as an S-phase or G2/M cell cycle arrest to the cancer cells, respectively. These were directly linked with a significant upregulation of mRNA expression of Caspase3 and Bax genes and an extreme downregulation of the mRNA expression of Bcl2, particularly in the combination treatment group, as compared to the untreated cells. Finally, ESA-NPs improved the effectiveness of cetuximab, strongly caused apoptotic and antiproliferative action with lower systemic toxicity, and could be suggested for the targeted administration of anticancer medications in various nanosystems.

Taraxasterol alleviates aflatoxin B1-induced liver damage in broiler chickens via regulation of oxidative stress, apoptosis and autophagy

Aflatoxin B1 (AFB1) is the most dangerous and abundant mycotoxin, which is toxic to almost all animals, and poultry is more sensitive to AFB1 toxicity. Ingesting AFB1-contaminated feed can cause significant liver damage and brings serious harm to poultry, which greatly restricts the development of the poultry industry. The present research was implemented to explore the intervention effect and its mechanism of taraxasterol on liver damage induced by AFB1 in broiler chickens. The liver damage model in broiler chickens was established by feeding 0.5 mg/kg AFB1 feed, and taraxasterol (25, 50 and 100 mg/kg BW, respectively) was given in the drinking water for 21 days. The growth performance, liver function, oxidative stress, apoptosis and autophagy were evaluated. The results showed that taraxasterol increased BW and reduced feed-to-gain ratio of broiler chickens induced by AFB1. Taraxasterol improved the levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyltransferase (GGT), total bilirubin (TBIL) and alkaline phosphatase (ALP), and attenuated hepatic histopathological changes induced by AFB1. Meantime, taraxasterol down-regulated cytochrome P450 (CYP450) enzyme system CYP1A1 and CYP2A6 mRNA expression, inhibited the overproduction of reactive oxygen species (ROS) and malondialdehyde (MDA), and enhanced the activities of antioxidant enzymes glutathione (GSH) and catalase (CAT) and the content of antioxidant superoxide dismutase (SOD) of the liver in broiler chickens induced by AFB1. Furthermore, taraxasterol up-regulated the mRNA and protein expression of hepatic nuclear factor E2 related factor 2 (Nrf2), heme oxygenase 1 (HO-1) and NAD(P)H: quinone oxidoreductase 1 (NQO1), and down-regulated the expression of hepatic kelch like ECH associated protein 1 (Keap1) induced by AFB1 in Keap1/Nrf2 signaling pathway. The ultrastructural observation and RT-qPCR results found that taraxasterol inhibited apoptosis of hepatocytes, up-regulated the expression of B-cell lymphoma-2 (Bcl-2) mRNA and down-regulated the expression of Bax and caspase3 mRNA. Further, taraxasterol restored the autophagy of hepatocytes and down-regulated the mRNA expression of phosphatidylinositol 3-kinase K (PI3K), protein kinase B (AKT) and mammalian target of rapamycin (mTOR) in AFB1-induced liver of broiler chickens. The above results indicate that taraxasterol alleviates liver damage induced by AFB1 in broiler chickens through regulation of Keap1/Nrf2 signaling pathway to exert its antioxidant effect, mitochondrial apoptosis pathway to improve anti-apoptotic ability and PI3K/AKT/mTOR pathway to restore autophagy.

Interactions of alcohol and combination antiretroviral (cART) drug in diabetic male Sprague Dawley rats: Hippocampal perturbations and toxicosis.

Hippocampal pathology in diabetes is constantly investigated but the resultant health impact of the concomitant presence of alcohol and combined antiretroviral therapy (cART) in diabetes requires further studies to delineate toxicities inimical to hippocampal normal function. Forty-eight male Sprague Dawley rats were divided into eight groups (n=6): negative control (NC), alcohol (AL), cART (AV), alcohol-cART (AA), diabetic control (DB), diabetes-alcohol (DAL), diabetes-cART (DAV), and diabetes-alcohol-cART (DAA) exposure groups. Following diabetes induction and sub-chronic (90 days) treatment exposure, hippocampal homogenates were profiled for pro-inflammatory cytokines and oxidative stress (MDA and GPx) using immunoassay, while apoptotic genes (BAX, Bcl2, and Caspase-3), insulin receptor genes (INSR and IRS-1), and blood-brain barrier (BBB) junctional proteins (claudin-5, and occludin) gene expression were assessed using qPCR. Histomorphology of hippocampal neuronal number, nuclei area, and volume of dentate gyrus and neurogenesis were accessed using Giemsa stain, Ki67, and DCX histochemistry respectively. A central hippocampal effect that underpins all treatments is the reduction of DG neuronal number and antioxidant (GPx), highlighting the venerability of the hippocampal dentate gyrus neurons to diabetes, alcohol, cART, and their combinatorial interactions. Additionally, elevated BAX, Bcl2, and IRS1 mRNA levels in the DAL group, and their downregulation in AA, suggests IRS-1-regulated apoptosis due to differential modulating effects of alcohol treatment in diabetes (DAL) in contrast to alcohol with cART (AA). Although the interaction in AA therapy ameliorated the independent alcohol and cART effects on MDA levels, pro-inflammatory cytokines, and DCX, the interaction in AA exacerbated a deficiency in the expression of INSR, IRS-1 (insulin sensitivity), and BBB mRNA which are implicated in the pathogenies of diabetes. Furthermore, the diabetic comorbidity groups (DAV, DAL, and DAA) all share a central effect of elevated hippocampal oxidative stress, BAX, and Caspase-3 mRNA expression with the reduced number of hippocampal neurons, dentate gyrus volume, and neurogenesis, highlighting neurodegenerative and cognitive deficiency implication of these comorbidity treatments. Considering these findings, assessment of hippocampal well-being in patients with these comorbidities/treatment combinations is invaluable and caution is advised particularly in alcohol use with cART prophylaxis in diabetes.