Solasonine alleviates high glucose-induced podocyte injury through increasing Nrf2-medicated inhibition of NLRP3 activation.

The worldwide high prevalence of diabetic nephropathy is one of the common causes of renal failure in diabetic patients. Hyperglycemia-caused podocyte injury is considered as a major contributor to diabetic kidney disease, accompanied by a chronic inflammatory condition. Pyroptosis, a characterized inflammatory form of programmed cell death, is believed to be involved in the pathogenesis of diabetic nephropathy. Solasonine (SS) is a natural alkaloid and received attention as a potential anticancer agent. However, its protective effect against hyperglycemia-caused podocyte injury remains to be determined. Our study found that SS alleviates cell apoptosis, and reduces pyroptosis and oxidative damage in high glucose (HG)-treated MPC5 podocytes. Pro-inflammatory cytokines, including interleukin (IL)-1ß and IL-18, and caspase-1 activity were markedly suppressed by SS in HG-treated MPC5 podocytes. SS also reduced HG-induced oxidative damage in MPC5 podocytes. Nrf2 expression was activated by SS in vitro under a HG condition. In addition, Nrf2 silencing attenuated the protective effect of SS against apoptosis, pro-inflammatory cytokines release, caspase-1 activity, and oxidative damage in MPC5 podocytes under a HG condition. Taken together, our findings revealed for the first time that SS alleviated high glucose-induced podocyte apoptosis, pyroptosis, and oxidative damage via regulating the Nrf2/NLRP3 signaling pathway. Our results indicate that SS has the potential as a therapeutic agent for podocyte injury in diabetic nephropathy.

Antifouling Zwitterionic Coating via Electrochemically Mediated Atom Transfer Radical Polymerization on Enzyme-Based Glucose Sensors for Long-Time Stability in 37 °C Serum.

In this study, a versatile fabrication method for coating enzyme-based biosensors with ultrathin antifouling zwitterionic polymer films to meet the challenge of the long-time stability of sensors in vivo was developed. Electrochemically mediated atom transfer radical polymerization (eATRP) was applied to polymerize zwitterionic sulfobetaine methacrylate monomers on the rough enzyme-absorbed electrode surfaces; meanwhile, a refined overall bromination was developed to improve the coverage of polymers on the biosensor surfaces and to maintain the enzyme activity simultaneously for the first time. X-ray photoelectron spectroscopy and atomic force microscopy were used to characterize the properties of the polymer layers. The antifouling performance and long-time stability in 37 °C undiluted bovine serum in vitro were evaluated. The results showed that the polymer brush coatings diminished over 99% nonspecific protein adsorption and that the sensitivity of the evaluated sensor was maintained at 94% after 15 days. The overall sensitivity deviation of 7% was nearly 50% lower than that of the polyurethane-coated ones and also much smaller than the current commercially available glucose biosensors. The results suggested that this highly controllable electrodeposition procedure could be a promising method to develop implantable biosensors with long-time stability.

Comprehensive Study of the Effects of Nanopore Structures on Enzyme Activity for the Enzyme Based Electrochemical Biosensors Based on Molecular Simulation.

Assembly of biocompatible nanostructures to retain the enzyme activity and improve the biocatalytic ability is a decisive factor for enhancing the performance of enzyme biosensors. However, there is still a lack of molecular level understandings of the physicochemical interaction mechanism at the interface of biosensor electrodes and enzymes. Here, for the first time at molecular level, the effects of two classic biosensor electrode materials with different electrical properties and morphologies and glucose oxidase (GOD) on retaining the enzyme conformation were analyzed by molecular dynamics simulation. First, for the immobilization of GOD, the interfaces of zinc oxide (ZnO) with different electrical properties and 10 nm diameter ZnO nanopore were studied. Then, to simulate the sensing process when electric voltages are applied, positively charged gold planes and 10 nm diameter gold nanopore were investigated as well. The results showed that the nanopore structure was confirmed to be well adapted for the enzyme conformation retaining compared to the plane structure for both ZnO and gold materials, and they almost fit well with the sensitivity measurement results from many previously reported experimental studies. This study also indicates that molecular modeling of the interactions between biomolecules and functional nanostructures is helpful for developing high performance enzyme nanobiosensors.

Apigenin exerts protective effect and restores ovarian function in dehydroepiandrosterone induced polycystic ovary syndrome rats: a biochemical and histological analysis.

BACKGROUND: Polycystic ovarian syndrome (PCOS) is one of the major causes encouraging the elevation of androgens, obesity along with menstrual complications. Here we study the effect of Apigenin in rat model of polycystic ovarian syndrome. METHODS: Female Sprague Dawley (SD) rats were treated with Dehydroepiandrosterone (DHEA) (6 mg/100g) opting the post-pubertal approach for developing rat model of polycystic ovarian syndrome, Metformin was used as standard. The treatments were given for 21 days along with coloproctological analysis. After the treatment regimen, the biochemical analysis was carried in plasma samples, whereas the ovaries were submitted for histopathological analysis. RESULTS: The treatment of DHEA resulted in disturbed lipid profile and anti-oxidant status along with increased weight, ovarian diameter and cysts in rats confirming the development of PCOS. However, treatment of Apigenin showed ameliorative effect by improving the lipid profile and anti-oxidant status, the treatment also normalised the body weight, reduced ovarian diameter, cysts and restored the healthy follicles compared to control rats. The treatment of Apigenin also suppressed the levels of oestradiol and testosterone compared to control group, also, levels of progesterone were increased in Apigenin treated group of rats. The treatment of Apigenin suppressed the levels of inflammatory cytokines TNF-α and IL-6. It was observed that the effect of Apigenin were to some extent parallel to standard drug Metformin. CONCLUSION: The findings confirmed that Apigenin ameliorates the disturbed hormonal levels, lipid profile and antioxidant status in PCOS rats.

Sp1-mediated upregulation of Prdx6 expression prevents podocyte injury in diabetic nephropathy via mitigation of oxidative stress and ferroptosis.

Glomerular podocyte damage is considered to be one of the main mechanisms leading to Diabetic nephropathy (DN). However, the relevant mechanism of podocyte injury is not yet clear. This study aimed to investigate the effect of peroxiredoxin 6 (Prdx6) on the pathogenesis of podocyte injury induced by high glucose (HG). The mouse glomerular podocyte MPC5 was stimulated with 30 nM glucose, and the Prdx6 overexpression vector or specificity protein 1 (Sp1) overexpression vector was transfected into MPC5 cells before the high glucose stimulation. As results, HG treatment significantly reduced the expression of Prdx6 and Sp1 in MPC5 cells. Prdx6 overexpression increased cell viability, while inhibited podocyte death, inflammation and podocyte destruction in HG-induced MPC5 cells. Prdx6 overexpression inhibited HG-induced ROS and MDA production, while restored SOD and GSH activity in MPC5 cells. Prdx6 overexpression also eliminated ferroptosis caused by HG, which was reflected in the suppression of iron accumulation and the increase in SLC7A11 and GPX4 expression. The improvement effect of Prdx6 on HG-induced podocyte damage could be eliminated by erastin. Moreover, Sp1 could bind to the three Sp1 response elements in the Prdx6 promoter, thereby directly regulating the transcriptional activation of Prdx6 in podocytes. Silencing Sp1 could eliminate the effect of Prdx6 on HG-induced podocyte damage. Further, Prdx6 overexpression attenuated renal injuries in streptozotocin-induced DN mice. Sp1-mediated upregulation of Prdx6 expression prevents podocyte injury in diabetic nephropathy via mitigation of oxidative stress and ferroptosis, which may provide new insights for the study of the mechanism of DN.

Synergistic and protective effect of atorvastatin and amygdalin against histopathological and biochemical alterations in Sprague-Dawley rats with experimental endometriosis.

The aim of the present study was to evaluate the protective effects of combined atorvastatin and amygdalin in a rat model of endometriosis. Tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), matrix metalloproteinase-2 (MMP-2) and MMP-9 levels in the peritoneal fluid were determined. The expression of TNF-α, IL-6, MMP-2, and MMP-9 mRNA, and the levels of lipid peroxidation, reduced glutathione (GSH), superoxide dismutase (SOD), catalase, and glutathione peroxidase (Gpx) were measured. Histopathological analysis was also conducted. The results showed that peritoneal TNF-α, IL-6, MMP-2, and MMP-9 levels were reduced by > 50%, and mRNA expression was decreased. Lipid peroxidation was considerably reduced, while GSH, SOD, Gpx, and catalase levels increased by > 40%. Reductions in leukocyte infiltration and fibrosis following treatment were also observed. Thus, our study suggested that combined treatment consisting of atorvastatin and amygdalin attenuates endometriosis. A detailed investigation of molecular mechanism of atorvastatin and amygdalin in endometriosis is needed.

Relationship between serum apolipoproteins levels and retinopathy risk in subjects with type 2 diabetes mellitus.

AIMS: Prognostic significance of apolipoproteins in diabetic retinopathy risk has not been well investigated. The aim of this study was to reveal the relationship between the risk of diabetic retinopathy and the levels of several apolipoproteins and their ratios in a 10-year prospective cohort. METHODS: A total of 1023 diabetic patients without retinopathy were selected from a 10-year hospital-based diabetic cohort. In this cohort, all subjects had type 2 diabetes. Blood samples were obtained, and serum levels of several apolipoproteins were measured. In the follow-up period, diabetic retinopathy was diagnosed by two ophthalmologists through a series of ophthalmologic examinations. A Cox proportional hazard analysis was adopted to determine the relationship between the risk of diabetic retinopathy and the levels of several apolipoproteins and their ratios. RESULTS: In the follow-up period, 315 diabetic patients were suffered from diabetic retinopathy, and the remaining 708 patients did not. Baseline serum level of apoAI ≥ 7.4 µmol/L was related to the decreased risk of diabetic retinopathy (HR 0.86, 95% CI 0.70-0.99). Baseline levels of apoCIII ≥ 6.3 µmol/L, apoE ≥ 1.1 µmol/L, apoCIII-to-apoAI ratio ≥ 0.9 and apoE-to-apoAI ratio ≥ 0.2 were associated with the increased risk of this complication (HR 1.25, 95% CI 1.04-1.49; HR 1.23, 95% CI 1.03-1.47; HR 1.34, 95% CI 1.11-1.60; HR 1.21, 95% CI 1.01-1.46). CONCLUSION: Elevated level of apoAI might be a protective factor for diabetic retinopathy. Increased levels of apoCIII, apoE, apoCIII-to-apoAI and apoE-to-apoAI ratios might be risk factors for this complication.

Functional expression of CXC chemokine recepter-4 mediates the secretion of matrix metalloproteinases from mouse hepatocarcinoma cell lines with different lymphatic metastasis ability.

CXC chemokine recepter-4 (CXCR4) and its ligand, stromal cell-derived factor-1alpha (SDF-1alpha) have been implicated in the organ-specific metastasis of several malignancies. Hca-F and its syngeneic cell line Hca-P are mouse hepatocarcinoma cell lines with high and low potential of lymphatic metastasis, respectively. Previous studies showed that the secretion of matrix metalloproteinases (MMPs) associated with the metastatic ability of Hca-F and Hca-P cell line depending on the lymph node environment. However, the mechanism of this process has remained unclear. This study investigated the roles of CXCR4 on Hca-F cell and SDF-1alpha of lymph node in lymphatic metastasis. The RT-PCR and Flow cytometry analysis results show that Hca-F cells express higher level CXCR4 mRNA and cell-surface CXCR4 protein, as compared with Hca-P cells. Treatment of recombinant SDF-1alpha proteins induced greater amount of calcium-flux in Hca-F cells than that in Hca-P cells, demonstrating higher functional CXCR4 expression on Hca-F cells than that on Hca-P cells. Furthermore, both the cell-free extratcs of lymph node and recombinant SDF-1alpha proteins induced secretions of active MMP-9 and MMP-2 from Hca-F cells in vitro. But those secretions were significantly reduced by blockade of cell surface CXCR4 with rabbit anti-mouse CXCR4 polyclonal antibody (pAb) and neutralization of SDF-1alpha in lymph node extracts with rabbit anti-mouse SDF-1alpha pAb as well. These results suggest that the CXCR4/SDF-1alpha system mediates active MMP-9 and MMP-2 secretion from Hca-F and Hca-P cells, which facilitates lymphogenous metastasis of those cells consequently.

A needle-type glucose biosensor based on PANI nanofibers and PU/E-PU membrane for long-term invasive continuous monitoring.

A minimally invasive glucose biosensor capable of continuous monitoring of subcutaneous glucose has been developed in this study. This sensor was prepared using electropolymerized conductive polymer polyaniline (PANI) nanofibers as an enzyme immobilization material and polyurethane (PU)/epoxy-enhanced polyurethane (E-PU) bilayer coating as a protective membrane. The sensor showed almost the same sensitivity (63nA/mM) and linearity (0-20mM with the correlation coefficient r2 of 0.9997) in both PBS and bovine serum tests. When stored in 37°C bovine serum, the sensor's sensitivity gradually increased about 30% of the initial value within the first 13 days and then remained stable for the rest of the study period of 53 days. In vivo implantation experiments using mice models showed real-time response to the variation of blood glucose with an average signal delay of about 8min. Continuous monitoring showed that the sensor response increased for the first 12 days and then entered a stable period for 14 days. The sensor's baseline (530±10nA) and the total response to 1ml 50% dextrose injection were almost the same (267±15nA) in the stable period. The in vivo stable performances indicated that the sensor could be used as an implantable device for long-term invasive monitoring of blood glucose.

Caveolin-1 up-regulates CD147 glycosylation and the invasive capability of murine hepatocarcinoma cell lines.

CD147 which is a regulator of matrix metalloproteinase (MMP) production on the surface of many malignant tumor cells, shows a highly specific association with caveolin-1 (Cav-1). As a result of heterogeneous N-glycosylation, CD147 exists in both highly glycosylated form, HG-CD147 ( approximately 40-60kDa) and lowly glycosylated form, LG-CD147 ( approximately 32kDa). This study investigated the possible role of Cav-1 in CD147 glycosylation in the HcaF, HcaP and Hepa1-6 mouse hepatocarcinoma cell lines, which have high, low and no metastatic potential in the lymph nodes, respectively, and in the normal mouse liver cell line IAR-20. Using an RNA interference (RNAi) strategy, we showed that the down-regulation of Cav-1 in Hca-F/RNAi cells could suppress the conversion of LG-CD147 to HG-CD147, down-regulate MMP-11 expression and decrease Hca-F/RNAi cell invasion. Conversely, a stable high expression of Cav-1 in Hepa1-6/Cav-1 cell could cause a specific increase of HG-CD147, up-regulate MMP-11 protein expression and enhance Hepa1-6/Cav-1 cell invasion. In conclusion, Cav-1 expression leads to an increased proportion of HG-CD147 relative to LG-CD147, increased production of MMP-11 and a higher invasive capability. Cav-1 is therefore proposed to act as both an oncogene and a tumor suppressor gene, and could represent a new potential target for gene therapy.

The fabrication of superlow protein absorption zwitterionic coating by electrochemically mediated atom transfer radical polymerization and its application.

A well-controllable electrochemically mediated surface-initiated atom transfer radical polymerization (e-siATRP) method for the fabrication of superlow protein absorption zwitterionic hydrogel coatings based on poly(sulbetaine methacrylate) (pSBMA) was developed in this work. The effects of the electric condition on polymerization as well as its antifouling performances both in vitro and in vivo were also investigated. Different potentials (-0.08 V, -0.15 V and -0.22 V) and polymerization times (from 8 to 48 h) were chosen to study the polymerization procedure. X-ray photoelectron spectroscopy, atomic force microscopy and ellipsometry measurements were used to characterize the properties of the polymer layers. Ellipsometry measurements showed that a higher potential provided faster polymerization and thicker polymer layers; however, the protein absorption experiments showed that the best polymerization condition was under a constant potential of -0.15 V and 32 h, under which the protein absorption was 0.8% in an enzyme-linked immunosorbent assay (compared to a bare gold electrode). The electrodes with a pSBMA coating effectively deduced the current sensitivity decay both in undiluted serum and in vivo. The usage of the commercially available polymerization monomer of SBMA, the simple convenient synthesis process regardless of the presence of oxygen and the excellent controllability of e-siATRP make it a very promising and universal technique in the preparation of zwitterionic polymer coatings, especially in the development of biocompatible material for implantable devices such as neural and biosensor electrodes.

Study of glucose biosensor lifetime improvement in 37°C serum based on PANI enzyme immobilization and PLGA biodegradable membrane.

Glucose sensors with long life span were fabricated and evaluated in 37°C serum to imitate in vivo conditions. Polyaniline nanofibers were electrodeposited on sensor electrodes for excessive glucose oxidase immobilization to extend the lifetime of sensors. The sensitivity of sensors stored in 37°C bovine serum was stable for 20 days without apparent descents, and still remained 45% of its initial value after 44 days. In order to further improve the life span of sensors, a degradable polymer film, poly lactic-co-glycolic acid (PLGA), was used as an out coating layer of glucose sensors. Results showed that the PLGA coated glucose sensors left 80% of its initial sensitivity after 44 days' storage in 37°C bovine serum. The sensors could be potentially used as long-term implantable devices for real time monitoring of glucose levels.

Fabrication and application of flexible graphene silk composite film electrodes decorated with spiky Pt nanospheres.

A free-standing graphene silk composite (G/S) film was fabricated via vacuum filtration of a mixed suspension of graphene oxide and silk fibres, followed by chemical reduction. Spiky structured Pt nanospheres were grown on the film substrate by cyclic voltammetry electrodeposition. The electrical and mechanical performance of a single graphene coated silk fibre was investigated. The conductivity of a single graphene coated silk fibre is 57.9 S m(-1). During 1000 bending measurements, the conductivity was stable and showed negligible variation. The G/S film has a sheet resistivity of 90 Ω â–¡(-1) with a porous and hierarchical structure. The spiky Pt nanosphere decorated G/S film was directly used as a H2O2 electrode with a sensitivity of 0.56 mA mM(-1) cm(-2), a linear range of 0-2.5 mM and an ultralow detection limit of 0.2 µM (S/N = 3). A glucose biosensor electrode was further fabricated by enzyme immobilization. The results show a sensitivity of 150.8 µA mM(-1) cm(-2) and a low detection limit of 1 µM (S/N = 3) for glucose detection. The strategy of coating graphene sheets on a silk fibre surface provides a new approach for developing electrically conductive biomaterials, tissue engineering scaffolds, bendable electrodes, and wearable biomedical devices.

Direct electron transfer glucose biosensor based on glucose oxidase self-assembled on electrochemically reduced carboxyl graphene.

A glucose biosensor based on direct electron transfer of glucose oxidase (GOD) self-assembled on the surface of the electrochemically reduced carboxyl graphene (ERCGr) modified glassy carbon electrode has been reported. X-ray photoelectron spectroscopy (XPS) analyses of ERCGr indicate most of the oxygen-containing groups such as epoxy/ether groups and hydroxyl groups in the carboxyl graphene were eliminated, while carboxylic acid groups remained. GOD was immobilized on the ERCGr modified glassy carbon electrode via self-assembly. The cyclic voltammetric result of the electrode shows a pair of well-defined and quasi-reversible redox peaks with a formal potential of -0.467 V and a peak to peak separation of 49 mV, revealing that the direct electron transfer between GOD and the electrode has been achieved. The proposed biosensor exhibits a linear response to glucose concentrations ranging from 2 to 18 mM with a detection limit of 0.02 mM. Moreover, this facile, fast, environment-friendly and economical preparation strategy of ERCGr may be extended for the preparation of other graphene based enzyme electrode biosensors.

Divergent expression and roles for caveolin-1 in mouse hepatocarcinoma cell lines with varying invasive ability.

Caveolin-1 is the major component protein of caveolae and associated with a lot of cellular events such as endocytosis, cholesterol homeostasis, signal transduction, and tumorigenesis. The majority of results suggest that caveolin-1 might not only act as a tumor suppressor gene but also a promoting metastasis gene. In this study, the divergent expression and roles of caveolin-1 were investigated in mouse hepatocarcinoma cell lines Hca-F, Hca-P, and Hepa1-6, which have high, low, and no metastatic potential in the lymph nodes, as compared with normal mouse liver cell line IAR-20. The results showed that expression of caveolin-1 mRNA and protein along with the amount of caveolae number in Hca-F cells was higher than that in Hca-P cells, but was not detectable in Hepa1-6 cells. When caveolin-1 expression in Hca-F cells was down-regulated by RNAi approach, Hca-F cells proliferation rate in vitro declined and the expression of lymphangiogenic factor VEGFA in Hca-F decreased as well. Furthermore, in vivo implantation assay indicated that reduction of caveolin-1 expression in Hca-F prevented the lymphatic metastasis tumor burden of Hca-F cells in 615 mice. These results suggest that caveolin-1 facilities the lymphatic metastasis ability of mouse hepatocarcinoma cells via regulation tumor cell growth and VEGFA expression.