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1.
World J Clin Cases ; 12(17): 3123-3129, 2024 Jun 16.
Article in English | MEDLINE | ID: mdl-38898829

ABSTRACT

BACKGROUND: Due to mechanical imbalance in the spine, elderly scoliosis patients tend to develop vertebral fracture nonunion, i.e., Kümmell disease, when osteoporotic vertebral compression fractures occur. However, accompanying vertebral rotational deformities make surgical procedures challenging risky. Such patients are usually compelled to undergo conservative treatment and there are very few reports on minimally invasive surgeries for them. We first-time report a patient with Kümmell disease and lumbar scoliosis treated with percutaneous kyphoplasty (PKP) under O-arm guidance. CASE SUMMARY: An 89-year-old female was admitted to the hospital due to delayed low back pain after a fall. She was diagnosed with Kümmell disease based on physical and radiologic examinations. The patient experienced severe scoliosis and subsequently underwent O-arm-guided kyphoplasty, resulting in a significant alleviation of low back pain. CONCLUSION: PKP has good efficacy in treating Kümmell disease. However, surgical risks are elevated in scoliosis patients with Kümmell disease due to the abnormal anatomical structure of the spine. O-arm assisted operations play a crucial role in decreasing surgical risks.

2.
Front Microbiol ; 15: 1394179, 2024.
Article in English | MEDLINE | ID: mdl-38881670

ABSTRACT

Conservation tillage (CT) is an important agronomic measure that facilitates soil organic carbon (SOC) accumulation by reducing soil disturbance and plant residue mulching, thus increasing crop yields, improving soil fertility and achieving C neutrality. However, our understanding of the microbial mechanism underlying SOC fraction accumulation under different tillage practices is still lacking. Here, a 6-year in situ field experiment was carried out to explore the effects of CT and traditional tillage (CK) practices on SOC fractions in an eolian sandy soil. Compared with CK, CT increased the particulate OC (POC) content in the 0-30 cm soil layer and the mineral-associated OC (MAOC) content in the 0-20 cm soil layer. Moreover, tillage type and soil depth had significant influences on the bacterial, fungal and protistan community compositions and structures. The co-occurrence network was divided into 4 ecological modules, and module 1 exhibited significant correlations with the POC and MOC contents. After determining their topological roles, we identified the keystone taxa in the network. The results indicated that the most common bacterial taxa may result in SOC loss due to low C use efficiency, while specific fungal (Cephalotrichum) and protistan (Cercozoa) species could facilitate SOC fraction accumulation by promoting macroaggregate formation and predation. Therefore, the increase in keystone fungi and protists, as well as the reduction in bacteria, drove module 1 community function, which in turn promoted SOC sequestration under CT. These results strengthen our understanding of microbial functions in the accrual of SOC fractions, which contributes to the development of conservation agriculture on the Northeast China Plain.

3.
Orthop Surg ; 15(10): 2647-2655, 2023 Oct.
Article in English | MEDLINE | ID: mdl-37652712

ABSTRACT

OBJECTIVE: The incidence of severe lower cervical fractures and dislocations due to trauma has increased significantly, and the optimal treatment remains controversial. This study compares the safety and efficacy of anterior cervical discectomy fusion (ACDF) combined with lateral mass screw (LMS) and with cervical pedicle screw (CPS) fixation surgery under O-arm navigation as single-stage treatments of severe lower cervical fracture dislocations. METHODS: Data from 48 patients who underwent ACDF + CPS (Group A) or ACDF + LMS (Group B) for severe lower cervical fracture dislocation between January 2016 and September 2020 were retrospectively reviewed. Groups A and B comprised 25 and 23 cases, respectively. Clinical parameters, such as operative time, intraoperative blood loss, number of fixed segments, posterior incision length, operative complications, and hospitalization days were recorded. Preoperatively and postoperatively, the sub-axial injury classification (SLIC) score, the American Spinal Injury Association (ASIA) impairment scale, and the Japanese Orthopaedic Association (JOA) score were recorded and analyzed using Student's t-test. RESULTS: The SLIC scores were 7.1 ± 1.2 and 7.5 ± 1.1, ASIA were 1.5 ± 0.6 and 1.2 ± 0.6, JOA score improvements were 3.2 ± 2.4 and 3.0 ± 2.1, operative times were 282.1 ± 91.7 and 266.5 ± 88.2 min, intraoperative blood losses were 437.8 ± 118.5 and 418.7 ± 104.2 mL, fixed segments were 2.8 ± 0.7 and 4.8 ± 1.1, and lengths of posterior incisions were 12.7 ± 2.8 and 13.8 ± 3.2 cm in Groups A and B, respectively. There was no significant difference between the two groups in the operative time, intraoperative blood loss, incision length, and postoperative recovery; however, group A had more fixed segments. At the final follow-up, no intraoperative or postoperative complications directly caused by the implant were present. Throughout the follow-up, all cases showed recovery and progressive improvement. CONCLUSION: Both ACDF + LMS and ACDF + CPS under O-arm navigation can safely and effectively restore cervical vertebral sequence, fully release spinal canal compression, and promote patients' neurological recovery. Thus, both are effective treatments for severe lower cervical fracture dislocations. However, compared to LMS, CPS under O-arm navigation has shorter fixed segments and induces less trauma.

4.
Mol Immunol ; 148: 54-67, 2022 08.
Article in English | MEDLINE | ID: mdl-35671559

ABSTRACT

Mastitis, an inflammation of the mammary gland, is a complex disease that affects the health of dairy cows worldwide. Sodium butyrate (SB) is a short-chain fatty acid that has recently been shown to have antioxidant, anti-inflammatory and anti-apoptotic potential in various cells types, although its role in bovine mammary epithelial cells (bMECs) has not been comprehensively reported. Therefore, the aim of this study was to assess the protective effect of sodium butyrate on Lipopolysaccharide (LPS)-induced mastitis model in vitro and to elucidate the possible underlying molecular mechanisms. The in vitro mastitis model was designed to investigate the regulatory effect of SB on LPS-induced inflammatory conditions in bMECs, with particular emphasis on oxidative stress, inflammatory response, apoptosis, and mitochondrial dysfunction. The results showed that SB co-treatment markedly prevented LPS-induced death of bMECs in a concentration-dependent manner. In addition, SB attenuated LPS-induced oxidative stress (OS) (Increased Intracellular ROS, MDA, and decreased SOD, GSH-Px and CAT activity), thereby reduced inflammation (increased expression of IL-6, IL-Iß, and TNF-α), and apoptosis (Increased the expression of caspases and Bax and decreased Bcl-2) via inhibiting NF-kB and caspase/bax signaling pathways. Furthermore, the protective effect of SB was also associated with the activation of endogenous antioxidant system (Nrf2, Keap1, NQO-1 and HO-1). Nrf2 silencing significantly abolished the protective effect of SB on bMECs. In conclusion, our findings suggest that SB has a significant protective effect on LPS-induced OS, inflammatory responses and apoptosis by activating Nrf2 and inhibiting NF-kB and ROS-mediated mitochondrial dysfunction. These results propose that SB may be an important regulator of OS and its subsequent inflammatory responses, and thus could be used as a therapeutic agent for bovine mastitis.


Subject(s)
Lipopolysaccharides , Mastitis , Animals , Apoptosis , Butyric Acid/metabolism , Butyric Acid/pharmacology , Butyric Acid/therapeutic use , Caspases/metabolism , Cattle , Epithelial Cells/metabolism , Female , Humans , Inflammation/drug therapy , Inflammation/metabolism , Kelch-Like ECH-Associated Protein 1/metabolism , Lipopolysaccharides/pharmacology , Mastitis/metabolism , NF-E2-Related Factor 2/metabolism , NF-kappa B/metabolism , Oxidative Stress , Reactive Oxygen Species/metabolism , bcl-2-Associated X Protein/metabolism
5.
In Vitro Cell Dev Biol Anim ; 57(5): 550-559, 2021 May.
Article in English | MEDLINE | ID: mdl-34081293

ABSTRACT

UFL1 is an ufmylation (a novel post-translational modification) E3 ligase, mainly located in the endoplasmic reticulum (ER), that has emerged as a significant regulator of several physiological and pathological processes. Yet its physiological function in milk synthesis in bovine mammary epithelial cells (BMECs) remains unknown. In this study, we investigated the effects of UFL1 in milk protein and fat synthesis-related gene expression, with a particular emphasis on the role of UFL1 in LPS-treated BMECs. Results showed that UFL1 depletion significantly reduced the expression of milk protein and fat synthesis-related gene and mTOR phosphorylation in both normal and LPS-treated BMECs. Overexpression of UFL1 enhanced the activation of the mTOR and milk protein and fat synthesis-related gene expression. Collectively, these above results strongly demonstrate that UFL1 could regulate milk protein and fat synthesis-related gene expression of BMECs probably via the mTOR signaling pathway.


Subject(s)
Glycolipids/biosynthesis , Glycoproteins/biosynthesis , Mammary Glands, Animal/metabolism , Milk Proteins/biosynthesis , Signal Transduction , TOR Serine-Threonine Kinases/metabolism , Ubiquitin-Protein Ligases/physiology , Animals , Cattle , Epithelial Cells/metabolism , Female , Flow Cytometry , Gene Expression Regulation , Lipid Droplets , Mammary Glands, Animal/cytology , Real-Time Polymerase Chain Reaction , Ubiquitin-Protein Ligases/metabolism
6.
Mol Immunol ; 135: 388-397, 2021 07.
Article in English | MEDLINE | ID: mdl-34022514

ABSTRACT

Heat stress-induced decline in milk production and mammary glands dysfunction are economically important challenges facing the dairy industry, especially in summer. Choline is an organic water-soluble compound that can regulate a series of vital biological process, including cellular structural integrity and oxidative stress. However, it is unclear whether choline plays an anti-apoptosis and antioxidant effect in heat stress-induced mammary epithelial cells. This study aimed to determine the antioxidant effect of choline on heat stress-induced apoptosis and oxidative stress and its underlying molecular mechanism in bovine mammary epithelial cells (MAC-T cells). The MAC-T cells were divided into four treatment groups: control (37℃), choline (37℃), heat stress (HS, 42℃), and HS + choline. The results showed that heat stress up-regulated the HSP70 and HSP90 expression both in mRNA and protein, enhanced ROS accumulation, increased malondialdehyde (MDA) content, reduced the superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) activity, significantly increased the expression of caspase-3 and upregulated the ratio of Bax/Bcl-2 and ultimately lead to oxidative stress and apoptosis in MAC-T cells. However, choline pretreatment reversed the above phenomenon compared with the HS group. The HS + choline group inhibited heat stress-induced phosphorylation of PERK, nuclear translocation of Nrf-2 and the protein expression of GRP78. In addition, the ratio of Bax/Bcl-2 and the expression of caspase-3 were significantly reduced in HS + choline group, thereby reduced the HS-induced oxidative stress and apoptosis in MAC-T cells. In conclusion, choline attenuates heat stress-induced oxidative stress and apoptosis of MAC-T cells by modulating PERK/Nrf-2 pathway.


Subject(s)
Apoptosis/drug effects , Choline/pharmacology , Heat-Shock Response/drug effects , NF-E2-Related Factor 2/metabolism , Oxidative Stress/drug effects , eIF-2 Kinase/metabolism , Animals , Antioxidants/pharmacology , Caspase 3/metabolism , Cattle , Cell Line , Endoplasmic Reticulum Chaperone BiP , Epithelial Cells/metabolism , Female , Glutathione Peroxidase/metabolism , HSP72 Heat-Shock Proteins/biosynthesis , HSP90 Heat-Shock Proteins/biosynthesis , Heat-Shock Proteins/metabolism , Mammary Glands, Animal/metabolism , Milk/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Reactive Oxygen Species/metabolism , Signal Transduction/drug effects , Superoxide Dismutase/metabolism , T-Lymphocytes/drug effects
7.
PLoS One ; 16(2): e0247559, 2021.
Article in English | MEDLINE | ID: mdl-33626085

ABSTRACT

The intramuscular fat (IMF) content and fatty acid composition are important meat quality traits that are mostly affected by the cattle breed. Muscle, adipose tissue and liver are important organs involved in the development of intramuscular adipose tissue. Thus, we hypothesized that there were marked differences in the adipogenesis and lipid metabolism of these tissues between Wagyu-cross and Holstein steers during the finishing phases. To test this hypothesis, we analyzed the expression levels of adipogenesis- and lipid metabolism-related genes in longissimus muscle (LM), subcutaneous fat (SCF) and liver from Wagyu-cross and Holstein steers at 26 months of age. The IMF content and fatty acid profile of LM were determined. Wagyu-cross steers had a higher IMF content and MUFA percentages in the LM than Holstein steers (P<0.05). The relative expression of FGF2, COL1A1, SREBP1c, SCD1, GRP78 and LEP was greater in the LM of Wagyu-cross steers than in Holstein steers (P<0.05). In contrast, Holstein steer SCF had higher (P<0.05) mRNA expression levels of FABP4 and ADIPOQ than Wagyu-cross steers. In the liver, the expression of SREBP1c and GRP78 in Wagyu-cross steers was significantly higher than that in Holstein steers (P<0.05). The results demonstrate that both intramuscular adipogenesis and fibrogenesis are enhanced in Wagyu-cross steers compared with Holstein steers during the finishing phase and that IMF deposition is positively correlated with the maturity of SCF and hepatic lipid accumulation in Wagyu-cross steers.


Subject(s)
Adipogenesis/genetics , Adipose Tissue/metabolism , Gene Expression , Lipid Metabolism/genetics , Liver/metabolism , Muscle, Skeletal/metabolism , Animals , Cattle , Fatty Acids/metabolism , Male , Species Specificity , Subcutaneous Fat/metabolism
8.
Life Sci ; 270: 119138, 2021 Apr 01.
Article in English | MEDLINE | ID: mdl-33524422

ABSTRACT

AIMS: Sodium propionate (SP) has been reported to possess an anti-inflammatory and anti-apoptotic potential by inhibiting certain signaling pathways and helps in reducing the pathological damages of the mammary gland. However, the effects of sodium propionate on attenuating Lipopolysaccharide (LPS)-induced inflammatory condition and cell damage in bovine mammary epithelial cells (bMECs) are not comprehensively studied yet. Therefore, the aim of the current investigation was to evaluate the protective effects of sodium propionate on LPS-induced inflammatory conditions and to clarify the possible underlying molecular mechanism in bMECs. MAIN METHODS: The effects of increasing doses of SP on LPS-induced inflammation, oxidative stress and apoptosis was studied in vitro. Furthermore, the underlying protective mechanisms of SP on LPS-stimulated bMECs was investigated under different experimental conditions. KEY FINDINGS: The results reveled that increased inflammatory cytokines, chemokines and those of tight junction's mRNA expression was significantly attenuated dose-dependently by propionate. Biochemical analysis revealed that propionate pretreatment modulated the LPS-induced intercellular reactive oxygen species (ROS) accumulation, oxidative and antioxidant factors and apoptosis rate. Furthermore, we investigated that the LPS activated nuclear factor-kB (NF-kB), caspase/Bax apoptotic pathways and Histone deacetylases (HDAC) was significantly attenuated by propionate in bMECs. SIGNIFICANCE: Our results suggest that sodium propionate is a potent agent for ameliorating LPS-mediated cellular disruption and limiting detrimental inflammatory responses, partly via maintaining blood milk barrier integrity, inhibiting HDAC activity and NF-kB signaling pathway.


Subject(s)
Milk/drug effects , Propionates/pharmacology , Animals , Anti-Inflammatory Agents/pharmacology , Apoptosis/drug effects , Cattle , Cells, Cultured , China , Cytokines/metabolism , Epithelial Cells/metabolism , Female , Inflammation/pathology , Lipopolysaccharides/pharmacology , Mammary Glands, Animal/drug effects , Mammary Glands, Animal/metabolism , Milk/metabolism , NF-kappa B/metabolism , Propionates/metabolism , Protective Agents/pharmacology , Signal Transduction/drug effects
9.
In Vitro Cell Dev Biol Anim ; 57(1): 66-75, 2021 Jan.
Article in English | MEDLINE | ID: mdl-33403623

ABSTRACT

The purpose of this study was to assess the effects of acetate and ß-hydroxybutyrate alone or in combination on lipogenic genes and their associated regulatory proteins in dairy cow mammary epithelial cells (DCMEC) using quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blotting, lipid droplet staining and a triglyceride content detection kit, to determine whether SCFA are related to milk fat synthesis regulation in DCMEC. Our experiment shows that addition of different concentrations of acetate, ß-hydroxybutyrate and their combinations to DCMEC increase in relative mRNA abundance of lipogenic genes and key transcription factors suggest an increase in lipogenic capacity, which is supported by an increased in cytosolic triglyceride content. Similarly, the protein expression level of acetyl-coenzyme A carboxylase (ACACA), fatty acid synthase (FASN) and sterol-coenzyme desaturase-1 (SCD1) genes and the transcription factor sterol regulatory element-binding protein-1 (SREBP1) were found to be increased by addition of acetate, ß-hydroxybutyrate and their combinations. The expression pattern of fat-related genes and proteins showed similar trends in almost all treatments, suggesting that common transcription factor are regulating these genes. These results show that acetate and ß-hydroxybutyrate regulate fat synthesis, further confirming that SCFAs work by targeting genes to activate the SREBP1 and insulin-induced gene 1 protein (INSIG1) signalling pathways in DCMEC.


Subject(s)
3-Hydroxybutyric Acid/pharmacology , Acetates/pharmacology , Epithelial Cells/metabolism , Gene Expression Regulation/drug effects , Lipid Droplets/metabolism , Lipogenesis/genetics , Mammary Glands, Animal/cytology , Triglycerides/metabolism , Animals , Cattle , Cell Survival/drug effects , Cell Survival/genetics , Dairying , Fatty Acids/biosynthesis , Female , Lipogenesis/drug effects , Milk/metabolism , Staining and Labeling
10.
Mol Immunol ; 127: 157-163, 2020 11.
Article in English | MEDLINE | ID: mdl-32987256

ABSTRACT

In the perinatal period of dairy cows, negative energy balance (NEB) is likely to occur, which increases the level of non-esterified fatty acids (NEFA) in the follicular fluid, hinders the proliferation of granulosa cells (GCs), and thus endangers the development of oocytes and the fecundity of dairy cows. We found that there were oxidative stress and inflammatory response in the serum of cows with perinatal ketosis. Whether the oxidative stress induced by NEFA is involved in the pyroptosis and inflammation of GCs remains unclear. After NEFA treatment, the expression of NLRP3 and caspase-1 and the release of inflammatory cytokines IL-1ß were increased in a dose-dependent manner, indicating that NEFA may contribute to pyroptosis. Besides, NEFA stimulation induced oxidative stress, resulting in the phosphorylation of NF-κB, and increased the production of interleukin (IL)-6 and nitric oxide (NO), indicating that NEFA may induce inflammation in GCs. However, the NEFA-mediated effects were observably reversed when the GCs were pre-treated with antioxidant and radical scavenger, N-acetylcysteine (NAC). Taken together, our results reveal that NEFA can induce pyroptosis and inflammation through NLRP3 inflammasome and TLR4/NF-κB pathway, respectively, and NAC can alleviate these conditions.


Subject(s)
Acetylcysteine/pharmacology , Fatty Acids, Nonesterified/toxicity , Granulosa Cells/pathology , Inflammation/pathology , Pyroptosis/drug effects , Animals , Cattle , Cell Survival/drug effects , Female , Granulosa Cells/drug effects , Inflammation Mediators/metabolism , Ketosis/pathology , Models, Biological , Oxidation-Reduction/drug effects , Oxidative Stress/drug effects , Up-Regulation/drug effects
11.
Calcif Tissue Int ; 107(5): 474-488, 2020 11.
Article in English | MEDLINE | ID: mdl-32767062

ABSTRACT

Wear particles released by joint implants are a major cause of osteolysis around the prosthesis by negatively affecting bone reconstruction. Bone marrow mesenchymal stem cells (BMMSCs) stimulated by wear particles showed an impaired osteogenic potential. Melatonin has been shown beneficial effects on intracellular antioxidant functions and bone formation; however, whether it could restore the osteogenic potential of BMMSCs inhibited by wear particles was unknown. This study aimed to evaluate the protective effect of melatonin on the osteogenic capacity of BMMSCs exposed to titanium (Ti) wear particles and to investigated the underlying mechanisms involving intracellular antioxidant properties. When BMMSCs were exposed to Ti particles in vitro, melatonin treatment successfully improved the matrix mineralization and expression of osteogenic markers in BMMSCs, while decreasing the levels of intracellular reactive oxygen species (ROS) and mitochondrial superoxide. The protective effect of melatonin on osteolysis was validated in a Ti particle-exposed murine calvarial model. Meanwhile, silent information regulator type 1 (SIRT1) and intracellular antioxidant enzymes were significantly up-regulated, particularly superoxide dismutase 2 (SOD2), in melatonin-treated BMMSCs. Furthermore, inhibition of SIRT1 by EX527 completely counteracted the protective effect of melatonin on Ti particle-treated BMMSCs, evidenced by the reduced expression of SOD2, increased ROS and superoxide, and decreased osteogenic differentiation. These results demonstrated that melatonin restored the osteogenic potential and improved the antioxidant properties of BMMSCs through the SIRT1 signaling pathway. Our findings suggest that melatonin is a promising candidate for treating osteolysis induced by wear particles.


Subject(s)
Melatonin/therapeutic use , Mesenchymal Stem Cells , Osteogenesis , Signal Transduction , Titanium/adverse effects , Animals , Bone Marrow Cells , Cell Differentiation , Cells, Cultured , Mice , Sirtuin 1/physiology , Superoxide Dismutase/physiology
12.
Oxid Med Cell Longev ; 2020: 4045674, 2020.
Article in English | MEDLINE | ID: mdl-32655766

ABSTRACT

Ubiquitin-like modifier 1 ligating enzyme 1 (UFL1) has been characterized as a ubiquitin-like (Ubl) protein that affects a range of cellular processes across various pathways. In this study, mouse mammary epithelial cells (HC11 cell line) and UFL1 knockout (KO) mice were used to establish UFL1 knockdown models to explore the influence of UFL1 on milk protein and fat synthesis in the mouse mammary gland and the underlying mechanisms. This is the first study to show UFL1 localization in mouse mammary epithelial cells. UFL1 depletion by transfected UFL1 siRNA (siUFL1) caused aggravated apoptosis. In addition, UFL1 depletion suppressed milk protein synthesis-related protein level in vivo and in vitro. Conversely, ACACA and FASN expressions increased in UFL1-deficient mice. Moreover, UFL1 depletion increased triglyceride synthesis levels and inhibited the p-JNK expression. Importantly, the expression of proteins related to milk protein synthesis was decreased in JNK- and UFL1-deficient cells, whereas proteins related to milk fat synthesis showed the opposite trend, indicating that UFL1 affects milk protein and fat synthesis via the suppression of JNK activation. Overall, our findings indicate that UFL1 plays a key role in mammary milk and fat synthesis via JNK activation.


Subject(s)
Fats/metabolism , JNK Mitogen-Activated Protein Kinases/metabolism , Mammary Glands, Animal/metabolism , Milk Proteins/biosynthesis , Ubiquitin-Protein Ligases/metabolism , Animals , Apoptosis/genetics , Cell Line , Epithelial Cells/metabolism , Female , Gene Expression Regulation , JNK Mitogen-Activated Protein Kinases/genetics , MAP Kinase Signaling System , Mice , Mice, Knockout , Milk Proteins/genetics , Triglycerides/metabolism , Ubiquitin-Protein Ligases/genetics
13.
Antioxidants (Basel) ; 9(6)2020 Jun 14.
Article in English | MEDLINE | ID: mdl-32545880

ABSTRACT

Negative energy balance (NEB) during the perinatal period can affect dairy cow follicular development and reduce the fecundity. Non-esterified fatty acid (NEFA) concentration is elevated during NEB, and is known to be toxic for multiple cell types. In the ovary, NEB increased NEFA, and may influences follicular growth and development. However, the effect and mechanism of NEFA on granulosa cells (GCs) in vitro remains unknown. In this study, we found that NEFA dose-dependently induced apoptosis in primary cultured granulosa cells. Mechanistically, our data showed that NEFA significantly increased reactive oxygen species (ROS) levels, resulting in the activation of endoplasmic reticulum stress (ERS) and eventually cell apoptosis in GCs. Moreover, NEFA also increased the phosphorylation levels of ERK1/2 and p38MAPK pathways, upregulated the expression of p53 and potentially promoted its translocation to the nuclear, thus transcriptionally activated Bax, a downstream gene of this pathway. NEFA also promoted nuclear factor E2 (Nrf2) expression and its level in the nuclear. To elucidate the mechanism of NEFA action, N-acetyl-L-cysteine (NAC), a ROS scavenger was used to verify the role of ROS in NEFA induced apoptosis of GCs. NAC pretreatment reversed the NEFA-induced ERS-related protein and apoptosis-related protein levels. Meanwhile, NAC pretreatment also blocked the phosphorylation of ERK1/2 and p38 induced by NEFA, and the nucleation of Nrf2 and p53, suggesting that ROS plays a crucial role in regulating the NEFA-induced apoptosis of GCs. Together, these findings provide an improved understanding of the mechanisms underlying GCs apoptosis, which could potentially be useful for improving ovarian function.

14.
Andrologia ; 52(7): e13600, 2020 Aug.
Article in English | MEDLINE | ID: mdl-32364254

ABSTRACT

Lead (Pb) is an environmental toxicant reported to impair male reproductive system. Betaine is a natural product which has promising beneficial effects against oxidative stress. In this experimental study, we evaluated the ameliorative effect of betaine on sperm quality and oxidative stress induced by lead (Pb) in the testis of adult male mice. Sixty male Kunming mice were divided equally into four groups: control group, betaine group (1% in drinking water), lead group (100 mg kg-1  bw-1  day-1 ) and betaine + lead group. In the last group, mice were supplemented with betaine for two weeks prior to the initiation of lead treatment and concurrently during lead treatment for 3 weeks until sacrificed. Our results indicated that in the lead-administrated group, body weights together with sperm count were significantly decreased (p < .05). The numbers of abnormal sperms were found to be higher in lead-treated mice. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (Cat) were significantly reduced, while the level of malondialdehyde (MDA) content was increased in the testis tissue following lead treatment. The mRNA levels of antioxidant-related genes (SOD1, GPX1 and CAT) were significantly decreased in the lead group. Betaine enhanced these parameters in betaine + lead group. In testis histology span, Johnson score was decreased (p < .05) in lead group and co-treatment with betaine increased Johnson score significantly in betaine + lead group. These results indicate that betaine improves sperm quality and ameliorate oxidative damage in testis of mice exposed to lead.


Subject(s)
Betaine , Testis , Animals , Antioxidants/metabolism , Antioxidants/pharmacology , Betaine/pharmacology , Catalase/metabolism , Glutathione Peroxidase/metabolism , Male , Malondialdehyde/metabolism , Mice , Oxidative Stress , Spermatozoa/metabolism , Superoxide Dismutase/metabolism , Testis/metabolism
15.
Biomolecules ; 10(2)2020 02 09.
Article in English | MEDLINE | ID: mdl-32050508

ABSTRACT

Ubiquitin-like modifier 1 ligating enzyme 1 (UFL1) is an E3 ligase of ubiquitin fold modifier 1 (UFM1), which can act together with its target protein to inhibit the apoptosis of cells. Lipopolysaccharides (LPS) can affect the ovarian health of female animals by affecting the apoptosis of ovarian granulosa cells. The physiological function of UFL1 on the apoptosis of bovine (ovarian) granulosa cells (bGCs) remains unclear; therefore, we focused on the modulating effect of UFL1 on the regulation of LPS-induced apoptosis in ovarian granulosa cells. Our study found that UFL1 was expressed in both the nucleus and cytoplasm of bGCs. The results here demonstrated that LPS caused a significant increase in the apoptosis level of bGCs in cows, and also dramatically increased the expression of UFL1. Furthermore, we found that UFL1 depletion caused a significant increase in apoptosis (increased the expression of BAX/BCL-2 and the activity of caspase-3). Conversely, the overexpression of UFL1 relieved the LPS-induced apoptosis. In order to assess whether the inhibition of bGCs apoptosis involved in the nuclear factor-κB (NF-κB) signaling pathway resulted from UFL1, we detected the expression of NF-κB p-p65. LPS treatment resulted in a significant upregulation in the protein concentration of NF-κB p-p65, and knockdown of UFL1 further increased the phosphorylation of NF-κB p65, while UFL1 overexpression significantly inhibited the expression of NF-κB p-p65. Collectively, UFL1 could suppress LPS-induced apoptosis in cow ovarian granulosa cells, likely via the NF-κB pathway. These results identify a novel role of UFL1 in the modulation of bGC apoptosis, which may be a potential signaling target to improve the reproductive health of dairy cows.


Subject(s)
Granulosa Cells/metabolism , NF-kappa B/metabolism , Ubiquitin-Protein Ligases/metabolism , Animals , Apoptosis/drug effects , Cattle , Cell Death/drug effects , Cells, Cultured , Female , Granulosa Cells/physiology , Lipopolysaccharides/pharmacology , Ovary/metabolism , Ovary/physiology , Signal Transduction/drug effects , Ubiquitin-Protein Ligases/physiology
16.
Animals (Basel) ; 10(1)2020 Jan 13.
Article in English | MEDLINE | ID: mdl-31941148

ABSTRACT

Zearalenone (ZEA) and T-2 are the most common mycotoxins in grains and can enter the animal and human food-chain and cause many health disorders. To elucidate the toxic response profile, we stimulated bovine granulosa cells (GCs) with ß-zearalenol or HT-2. Using isobaric tags for relative and absolute quantification (iTRAQ)-based proteomic, 178 and 291 differentially expressed proteins (DEPs, fold change ≥ 1.3 and p-value < 0.05) in ß-zearalenol and HT-2 groups were identified, respectively. Among these DEPs, there were 66 common DEPs between ß-zearalenol and HT-2 groups. These 66 DEPs were associated with 23 biological processes terms, 14 molecular functions terms, and 19 cellular components terms. Most heat shock proteins (HSPs) were involved in the toxic response. Reactive oxygen species accumulation, the endoplasmic reticulum (ER)-stress related marker molecule (GRP78), and apoptosis were activated. ß-zearalenol and HT-2 inhibited oestradiol (E2) production. These results emphasized the important function of HSPs, clarified oxidative stress, and demonstrated the caspase-3 signaling cascade involved in mycotoxin-treated toxic response, along with decreased E2 production. This study offers new insights into the toxicity of ß-zearalenol and HT-2 on ovarian granulosa cells.

17.
Cell Stress Chaperones ; 25(2): 223-233, 2020 03.
Article in English | MEDLINE | ID: mdl-31925678

ABSTRACT

Bovine mastitis is a common inflammatory disease caused by various factors. The main factor of mastitis is pathogenic microorganism infection, such as Staphylococcus aureus, Escherichia coli, and Streptococcus. Cronobacter sakazakii (C. sakazakii) is a newly discovered pathogenic bacteria in milk products, which seriously threat human health in recent years. At present, it has not been reported that the pathogenesis of mastitis is caused by C. sakazakii. This study investigated the inflammation of mammary gland epithelium, which was induced by C. sakazakii for the first time. We focused on bacterial isolation, histological observation, AIM2 inflammasome pathways, endoplasmic reticulum stress, and apoptosis. The results showed that C. sakazakii-induced inflammation caused damage of tissue, significantly increased the production of pro-inflammatory cytokines (including TNF-α, IL-1ß, and IL-6), activated the AIM2 inflammasome pathway (increased the expression of AIM2 and cleaved IL-1ß), and induced endoplasmic reticulum stress (increased the expression of ERdj4, Chop, Grp78) and apoptosis (increased the ratio of Bax/Bcl-2, a marker of apoptosis). In conclusion, it is suggested that it maybe inhibite AIM2 inflammasome pathways and alleviate endoplasmic reticulum stress (ER stress) against the C. sakazakii-induced inflammation.


Subject(s)
Cronobacter sakazakii , DNA-Binding Proteins/metabolism , Endoplasmic Reticulum Stress , Enterobacteriaceae Infections/metabolism , Inflammation/metabolism , Mastitis, Bovine , Milk/microbiology , Animals , Cattle , Cell Line , Cronobacter sakazakii/immunology , Cronobacter sakazakii/isolation & purification , Cytokines/metabolism , Endoplasmic Reticulum Chaperone BiP , Epithelial Cells , Female , Mastitis, Bovine/immunology , Mastitis, Bovine/microbiology , Mice , Mice, Inbred C57BL
18.
Cell Stress Chaperones ; 24(6): 1115-1125, 2019 11.
Article in English | MEDLINE | ID: mdl-31721015

ABSTRACT

Ubiquitin-fold modifier 1 (UFM1)-specific ligase 1 (UFL1) is an important component of the UFM1 conjugation system, which is required for various cellular processes including protein translation, apoptosis, autophagy, and signal transduction. However, both, the expression of UFL1 in mammary cells and its role in endoplasmic reticulum (ER) stress in bovine mammary epithelial cells (BMECs) remain to be fully elucidated. Here, we characterized the potential roles of UFL1 in BMECs. Amino acid sequence comparison indicated that bovine UFL1 shares a high level of sequence identity with the UFL1 of other ruminant species. Notably, UFL1 expression in BMECs was increased by endoplasmic reticulum (ER) stress induced by treatment with tunicamycin (TM). ER stress-related gene expression was further increased in UFL1 knockdown cells upon TM treatment. Moreover, UFL1 overexpression inhibited TM-stimulated ER stress and alleviated ER stress-induced autophagy. Together, our results indicated that UFL1 is a novel ER stress-responsive protein in BMECs. Thus, our study provides a basis for further research into ER stress-related processes in bovine mammary tissues and potential targets for alleviating ER stress in these cells.


Subject(s)
Endoplasmic Reticulum Stress/physiology , Endoplasmic Reticulum/metabolism , Epithelial Cells/metabolism , Mammary Glands, Animal/metabolism , Ubiquitin-Protein Ligases/physiology , Animals , Autophagy/physiology , Cattle , Cells, Cultured , Epithelial Cells/cytology , Female , Mammary Glands, Animal/cytology , Tunicamycin/pharmacology
19.
J Dairy Res ; 86(4): 416-424, 2019 Nov.
Article in English | MEDLINE | ID: mdl-31722754

ABSTRACT

Mastitis, a major infectious disease in dairy cows, is characterized by an inflammatory response to pathogens such as Escherichia coli and Staphylococcus aureus. To better understand the immune and inflammatory response of the mammary gland, we stimulated bovine mammary gland epithelial cells (BMECs) with E. coli-derived lipopolysaccharide (LPS). Using transcriptomic and proteomic analyses, we identified 1019 differentially expressed genes (DEGs, fold change ≥2 and P-value < 0.05) and 340 differentially expressed proteins (DEPs, fold change ≥1.3 and P-value < 0.05), of which 536 genes and 162 proteins were upregulated and 483 genes and 178 proteins were downregulated following exposure to LPS. These differentially expressed genes were associated with 172 biological processes; 15 Gene Ontology terms associated with response to stimulus, 4 associated with immune processes, and 3 associated with inflammatory processes. The DEPs were associated with 51 biological processes; 2 Gene Ontology terms associated with response to stimulus, 1 associated with immune processes, and 2 associated with inflammatory processes. Meanwhile, several pathways involved in mammary inflammation, such as Toll-like receptor, NF-κB, and NOD-like receptor signaling pathways were also represented. NLRP3 depletion significantly inhibited the expression of IL-1ß and PTGS2 by blocking caspase-1 activity in LPS-induced BMECs. These results suggest that NLR signaling pathways works in coordination with TLR4/NF-κB signaling pathways via NLRP3-inflammasome activation and pro-inflammatory cytokine secretion in LPS-induced mastitis. The study highlights the function of NLRP3 in an inflammatory microenvironment, making NLRP3 a promising therapeutic target in Escherichia coli mastitis.


Subject(s)
Epithelial Cells/metabolism , Inflammation/metabolism , Lipopolysaccharides/toxicity , Mammary Glands, Animal/cytology , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Animals , Cattle , Cells, Cultured , Epithelial Cells/drug effects , Female , Gene Expression Regulation/drug effects , Inflammation/genetics , NLR Family, Pyrin Domain-Containing 3 Protein/genetics , Proteomics , Transcriptome
20.
Front Immunol ; 10: 1579, 2019.
Article in English | MEDLINE | ID: mdl-31447828

ABSTRACT

Bovine mastitis is a common inflammatory disease caused by multiple factors in early lactation or dry period. Genome wide association studies (GWAS) can provide a convenient and effective strategy for understanding the biological basis of mastitis and better prevention. 2b-RADseq is a high-throughput sequencing technique that offers a powerful method for genome-wide genetic marker development and genotyping. In this study, single nucleotide polymorphisms (SNPs) of the immune-regulated gene correlative with mastitis were screened and identified by two stage association analysis via GWAS-2b-RADseq in Chinese Holstein cows. We have screened 10,058 high quality SNPs from 7,957,920 tags and calculated their allele frequencies. Twenty-seven significant SNPs were co-labeled in two GWAS analysis models [Bayesian (P < 0.001) and Logistic regression (P < 0.01)], and only three SNPs (rs75762330, C > T, PIC = 0.2999; rs88640083, A > G, PIC = 0.1676; rs20438858, G > A, PIC = 0.3366) were annotated to immune-regulated genes (PTK2B, SYK, and TNFRSF21). Identified three SNPs are located in non-coding regions with low or moderate genetic polymorphisms. However, independent sample population validation (Case-control study) data showed that three important SNPs (rs75762330, P < 0.025, OR > 1; rs88640083, P < 0.005, OR > 1; rs20438858, P < 0.001, OR < 1) were significantly associated with clinical mastitis trait. Importantly, PTK2B and SYK expression was down-regulated in both peripheral blood leukocytes (PBLs) of clinical mastitis cows and in vitro LPS (E. coli)-stimulated bovine mammary epithelial cells, while TNFRSF21 was up-regulated. Under the same conditions, expression of Toll-like receptor 4 (TLR4), AKT1, and pro-inflammatory factors (IL-1ß and IL-8) were also up-regulated. Interestingly, network analysis indicated that PTK2B and SYK are co-expressed in innate immune signaling pathway of Chinese Holstein. Taken together, these results provided strong evidence for the study of SNPs in bovine mastitis, and revealed the role of SYK, PTK2B, and TNFRSF21 in bovine mastitis susceptibility/tolerance.


Subject(s)
Focal Adhesion Kinase 2/physiology , Genome-Wide Association Study , Mastitis, Bovine/genetics , Polymorphism, Single Nucleotide , Receptors, Tumor Necrosis Factor/physiology , Syk Kinase/physiology , Animals , Cattle , Female , Focal Adhesion Kinase 2/genetics , Genetic Predisposition to Disease , Mastitis, Bovine/etiology , Mastitis, Bovine/immunology , Receptors, Tumor Necrosis Factor/genetics , Syk Kinase/genetics
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