Dietary additive ferulic acid alleviated oxidative stress, inflammation, and apoptosis induced by chronic exposure to avermectin in the liver of common carp (Cyprinus carpio).

Avermectin (AVM) has been utilized extensively in agricultural production since it is a low-toxicity pesticide. However, the pollution caused by its residues to fisheries aquaculture has been neglected. As an abundant polyphenolic substance in plants, ferulic acid (FA) possesses anti-inflammatory and antioxidant effects. The goal of the study is to assess the FA's ability to reduce liver damage in carp brought on by AVM exposure. Four groups of carp were created at random: the control group; the AVM group; the FA group; and the FA + AVM group. On day 30, and the liver tissues of carp were collected and examined for the detection of four items of blood lipid as well as the activity of the antioxidant enzymes catalase (CAT), glutathione (GSH) and malondialdehyde (MDA) in carp liver tissues by biochemical kits, and the transcript levels of indicators of oxidative stress, inflammation and apoptosis by qPCR. The results showed that liver injury, inflammation, oxidative stress, and apoptosis were attenuated in the FA + AVM group compared to the AVM group. In summary, dietary addition of FA could ameliorate the hepatotoxicity caused by AVM in carp by alleviating oxidative stress, inflammation, apoptosis in liver tissues.

Wine- and stir-frying processing of Cuscutae Semen enhance its ability to alleviate oxidative stress and apoptosis via the Keap 1-Nrf2/HO-1 and PI3K/AKT pathways in H2O2-challenged KGN human granulosa cell line.

BACKGROUND: Cuscutae Semen (CS) has been prescribed in traditional Chinese medicine (TCM) for millennia as an aging inhibitor, an anti-inflammatory agent, a pain reliever, and an aphrodisiac. Its three main forms include crude Cuscutae Semen (CCS), wine-processed CS (WCS), and stir-frying-processed CS (SFCS). Premature ovarian insufficiency (POI) is a globally occurring medical condition. The present work sought a highly efficacious multi-target therapeutic approach against POI with minimal side effects. Finally, it analyzed the relative differences among CCS, WCS and SFCS in terms of their therapeutic efficacy and modes of action against H2O2-challenged KGN human granulosa cell line. METHODS: In this study, ultrahigh-performance liquid chromatography (UPLC)-Q-ExactiveTM Orbitrap-mass spectrometry (MS), oxidative stress indices, reactive oxygen species (ROS), Mitochondrial membrane potential (MMP), real-time PCR, Western blotting, and molecular docking were used to investigate the protective effect of CCS, WCS and SFCS on KGN cells oxidative stress and apoptosis mechanisms. RESULTS: The results confirmed that pretreatment with CCS, WCS and SFCS reduced H2O2-induced oxidative damage, accompanied by declining ROS levels and malondialdehyde (MDA) accumulation in the KGN cells. CCS, WCS and SFCS upregulated the expression of antioxidative levels (GSH, GSH/GSSG ratio, SOD, T-AOC),mitochondrial membrane potential (MMP) and the relative mRNA(Nrf2, Keap1, NQO-1, HO-1, SOD-1, CAT). They inhibited apoptosis by upregulating Bcl-2, downregulating Bax, cleaved caspase-9, and cleaved caspase-3, and lowering the Bax/Bcl-2 ratio. They also exerted antioxidant efficacy by partially activating the PI3K/Akt and Keap1-Nrf2/HO-1 signaling pathways. CONCLUSIONS: The results of the present work demonstrated the inhibitory efficacy of CCS, WCS and SFCS against H2O2-induced oxidative stress and apoptosis in KGN cells and showed that the associated mechanisms included Keap1-Nrf2/HO-1 activation, P-PI3K upregulation, and P-Akt-mediated PI3K-Akt pathway induction.

Scutellarin prevents acute alcohol-induced liver injury via inhibiting oxidative stress by regulating the Nrf2/HO-1 pathway and inhibiting inflammation by regulating the AKT, p38 MAPK/NF-κB pathways. / ç¯ç›èŠ±ä¹™ç´ é€šè¿‡è°ƒèŠ‚Nrf2/HO-1通路抑制氧化应激以及通过调节AKT、p38 MAPK/NF-κBé€šè·¯æŠ‘åˆ¶ç‚Žç—‡ååº”é¢„é˜²æ€¥æ€§é ’ç²¾æ€§è‚æŸä¼¤.

Alcoholic liver disease (ALD) is the most frequent liver disease worldwide, resulting in severe harm to personal health and posing a serious burden to public health. Based on the reported antioxidant and anti-inflammatory capacities of scutellarin (SCU), this study investigated its protective role in male BALB/c mice with acute alcoholic liver injury after oral administration (10, 25, and 50 mg/kg). The results indicated that SCU could lessen serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels and improve the histopathological changes in acute alcoholic liver; it reduced alcohol-induced malondialdehyde (MDA) content and increased glutathione peroxidase (GSH-Px), catalase (CAT), and superoxide dismutase (SOD) activity. Furthermore, SCU decreased tumor necrosis factor-|α (TNF-|α), interleukin-6 (IL-6), and IL-|1ß messenger RNA (mRNA) expression levels, weakened inducible nitric oxide synthase (iNOS) activity, and inhibited nucleotide-binding oligomerization domain (NOD)|-like receptor protein 3 (NLRP3) inflammasome activation. Mechanistically, SCU suppressed cytochrome P450 family 2 subfamily E member 1 (CYP2E1) upregulation triggered by alcohol, increased the expression of oxidative stress-related nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) pathways, and suppressed the inflammation-related degradation of inhibitor of nuclear factor-|κB (NF-|κB)|-|α (IκBα) as well as activation of NF|-|κB by mediating the protein kinase B (AKT) and p38 mitogen-activated protein kinase (MAPK) pathways. These findings demonstrate that SCU protects against acute alcoholic liver injury via inhibiting oxidative stress by regulating the Nrf2/HO-1 pathway and suppressing inflammation by regulating the AKT, p38 MAPK/NF-κB pathways.

Protective Effects of Scutellarin on Acute Alcohol Intestinal Injury.

The present study aims to investigate the roles of scutellarin (SCU) on acute alcohol intestinal injury. Mice were divided into six groups: alcohol, three administration, negative control and positive drug bifendate control. The administration group mice were intraperitoneally injected with SCU for 3 consecutive days followed by alcohol gavage at an interval of 1 h. After the mice were sacrificed, colon tissue damage was evaluated by histopathological examination; the activities of inducible nitric oxide synthase (iNOS) and catalase (CAT), as well as the content of malondialdehyde (MDA) were detected using biochemical kits; the levels of inflammatory cytokines mRNA were determined by real-time fluorescence quantitative PCR; the protein expression levels of hemeoxygenase-1 (HO-1) and phosphorylated nuclear factor-ĸB p65 were measured via western blotting. The results showed that alcohol induced severe colon morphological degradation, epithelia atrophy, and more inflammatory cells infiltration in the submucosa. SCU treatment prevented this process, especially in the middle and high dose groups. Alcohol treatment caused excessive lipid peroxidation product accumulation of MDA, restrained the activity of antioxidant enzyme CAT, induced HO-1 expression in the colon, whereas low dose SCU treatment significantly down-regulated the MDA level, enhanced the CAT level, and accelerated HO-1 signals. SCU prevented alcohol stimulation triggered inflammatory response in colon tissues through significantly downregulating the iNOS activity, transcript levels of Tnf-α, Il-1ß and Il-6, and phosphorylation levels of NF-κB p65. These findings suggest that SCU protects the colon via antioxidant and anti-inflammatory mechanisms, making it a promising drug against alcohol-induced colon damage.

Hydrogen-Bonded Films for Zero-Order Release of Leuprolide.

Leuprolide has been widely used in androgen deprivation therapy for the treatment of advanced prostate cancer, but its use is still limited due to its short half-life. Herein, hydrogen-bonded layer-by-layer films are fabricated from PEGylated leuprolide (PEG-LEU) and tannic acid (TA). Because of its dynamic nature, the film disintegrates gradually in water and releases PEG-LEU and TA. The in vitro release profile indicated perfect zero-order kinetics, which is explained by the unique release mechanism. When implanted subcutaneously in male rats, the films maintain a constant serum drug level. For a 60-bilayer film, the serum drug level is maintained constant for ≈24 days. No initial burst release is observed, suggesting that the in vivo release also follows zero-order kinetics. Initially, an increase in the level of serum testosterone is induced by the released drug, followed by testosterone suppression to a constant level below the castrate level, which could be maintained as long as a constant serum drug level is maintained. Since the new drug carriers avoid an initial burst release of the drug and maintain a constant serum drug level and hence a constant serum testosterone level below the castrate level, these carriers are highly promising for androgen deprivation therapy.

PEGylated leuprolide with improved pharmacokinetic properties.

Leuprolide, a gonadotropin-releasing hormone (GnRH) agonist widely used in androgen deprivation therapy for the treatment of advanced prostate cancer, suffers from a short circulating half-life like other peptide therapeutics. As an attempt to improve its pharmacokinetic properties, two PEGylated leuprolides with different molecular weight were synthesized utilizing N-hydroxysuccinimidyl (NHS) conjugation chemistry. The reaction conditions, including reaction temperature, reaction time and feed ratio of the reactants, were optimized to obtain a higher yield. Reverse-phase high performance liquid chromatography (RP-HPLC) characterization indicates a high purity of the resulting conjugates. Matrix-assisted laser desorption mass spectrometry (MALDI-MS) characterization suggests a 1:1 PEGylation. 1H NMR study reveals that the reaction occurs on the imidazolyl group on the histidine residue and the conjugates are stable in pH7.4 aqueous solutions. The in vitro bioactivity of the conjugates was evaluated using both hormone-sensitive and hormone-insensitive cell lines. It was found that the PEGylated peptides can still counteract the stimulatory action of androgens and the mitogenic action of epidermal growth factor on cell proliferation. The in vivo bioactivity of the conjugates was also tested. Like the unmodified peptide, administration of the conjugates to male rats leads to an initial testosterone surge, followed by a suppression of testosterone secretion. Pharmacokinetics of the drugs after i.v. and s.c. administrations were determined. In both cases, a prolonged circulating half-life, an increased AUC, and a decreased Cl_F were observed for the PEGylated drugs.

Injectable Carrier for Zero-Order Release of Salmon Calcitonin.

Salmon calcitonin (sCT) was developed as an antiresorptive for the management of osteoporosis, a major public health threat worldwide. However, its clinical application was severely limited by its short half-life. Herein, an injectable drug carrier, that is, polylactic acid (PLA) microspheres coated with TA/PEG-sCT (TA: tannic acid. PEG-sCT: PEGylated sCT) layer-by-layer (LBL) films, was designed. An in vitro test demonstrated that, unlike previously developed drug carriers, the new carrier released PEG-sCT at a constant rate. The unique zero-order release kinetics originates from its unique drug release mechanism, that is, drug release via gradual disintegration of the dynamic TA/PEG-sCT LBL film. The small size of the PLA microspheres allows the carrier to be administrated via subcutaneous injection. An in vivo test demonstrated that a single injection of the carrier could maintain the plasma level of PEG-sCT stable for an extended period and thus induced a stable reduction in the plasma calcium level in rats. Using a rat model of osteoporosis induced by ovariectomy, it was further demonstrated that a single injection of the new carrier gave better therapeutic outcomes than daily injection of sCT of the same dose, thanks to the improved pharmacokinetic profile. Given the advantages of the new carrier, including facile subcutaneous administration, less frequent dosing, no initial burst release, no peak plasma drug level, and improved therapeutic outcomes, it is expected to have potential in long-term management of osteoporosis and other metabolic bone diseases.

Synthesis and anti-cancer evaluation of folic acid-peptide- paclitaxel conjugates for addressing drug resistance.

The drug resistance and the poor water solubility are major limitations of paclitaxel (PTX) of based chemotherapy. To conquer the two problems, targeting folate (FA) receptor PTX-lytic peptides conjugates were synthesized and evaluated. Compared with PTX, FA-P3-PTX and FA-P7-PTX displayed significantly enhanced cell toxicity in many cancer cells, particularly drug resistant cancer cells MCF-7/PTX. FA-P7-PTX possessed stronger effect on cell toxicity (IC50 = 2.92 ±â€¯0.2 µM), membrane disrupting activity and pro-apoptosis in MCF-7/PTX cells than FA-P3-PTX. Further investigation displayed that the anti-cancer mechanisms of FA-P3-PTX and FA-P7-PTX might be a mitochondrial impairment and caspase-3-dependent apoptotic cell death. Furthermore, the in vivo antitumor efficacy study confirmed that FA-P7-PTX performed more stronger potency in inhibition of tumors growth than PTX. The study demonstrated that conjugate FA-P7-PTX with superior properties for antineoplastic activity, which makes it a promising potential candidate for drug-resistant cancer therapy.

Injectable self-assembled peptide hydrogels for glucose-mediated insulin delivery.

Closed-loop glucose-responsive insulin delivery with excellent biocompatibility has the potential to improve the health and quality of life of diabetic patients. Herein, we developed an excellent glucose-responsive insulin delivery system using a pH-sensitive peptide hydrogel loaded with insulin and a glucose-specific enzyme. The designed peptide can be used as a carrier that is loaded with insulin and enzyme via a self-assembly process under physiological conditions. When hyperglycemia is encountered, the enzymatic conversion of glucose into gluconic acid leads to a decrease in the local pH, and the hydrogel is disassembled because of the strong inter- and intramolecular electrostatic repulsions between ornithine (Orn) residues; this is followed by the release of insulin. The glucose-responsive hydrogel system was characterized by studying its structure, conformation, rheology, morphology, acid sensitivity and the amounts of consistent release of insulin in vitro and in vivo. In vivo experiments indicated that the closed-loop insulin glucose-responsive system could efficiently regulate blood glucose in streptozocin-induced (STZ-induced) type 1 diabetic rats for 8 days.

pH-sensitive peptide hydrogel for glucose-responsive insulin delivery.

Glucose-responsive system is one of important options for self-regulated insulin delivery to treat diabetes, which has become an issue of great public health concern in the world. In this study, we developed a novel and biocompatible glucose-responsive insulin delivery system using a pH-sensitive peptide hydrogel as a carrier loaded with glucose oxidase, catalase and insulin. The peptide could self-assemble into hydrogel under physiological conditions. When hypoglycemia is encountered, neighboring alkaline amino acid side chains are significantly repulsed due to reduced local pH by the enzymatic conversion of glucose into gluconic acid. This is followed by unfolding of individual hairpins, disassembly and release of insulin. The glucose-responsive hydrogel system was characterized on the basis of structure, conformation, rheology, morphology, acid-sensitivity and the amount of consistent release of insulin in vitro and vivo. The results illustrated that our system can not only regulate the blood glucose levels in vitro but also in mice models having STZ-induced diabetes. STATEMENT OF SIGNIFICANCE: In this report, we have shown the following significance supported by the experimental results. 1. We successfully developed, characterized and screened a novel pH-responsive peptide. 2. We successfully developed a novel and biocompatible pH-sensitive peptide hydrogel as glucose-responsive insulin delivery system loaded with glucose oxidase, catalase and insulin. 3. We successfully confirmed that the hydrogel platform could regulate the blood glucose level in vitro and in vivo. Overall, we have shown enough significance and novelty with this smart hydrogel platform in terms of biomaterials, peptide chemistry, self-assembly, hydrogel and drug delivery. So we believe this manuscript is suitable for Acta Biomaterialia.

The Effects of an Empowerment Intervention on Renal Transplant Recipients: A Randomized Controlled Trial.

BACKGROUND: Renal transplantation is a vital treatment for end-stage renal disease. To help improve quality of life after renal transplant surgery, interventions are needed to strengthen the coping skills and self-care behaviors of patients. However, most research studies on self-care after renal transplantation have addressed related factors. Few studies have examined the effects of interventions on renal transplant recipients. PURPOSE: This study investigated the effects of an empowerment support group on the empowerment levels and self-care behaviors of renal transplant recipients. METHODS: This study was a randomized controlled trial. Eligible participants were individuals who had undergone a renal transplant within the past 20 years, were 18 years old or older, were able to read and write in Chinese, and were willing to participate. We recruited 122 renal transplant recipients from two medical centers in southern Taiwan. The renal transplant outpatients were randomly assigned into empowerment support (n = 56) and comparison (n = 66) groups. The developed measures as well as the content, protocols, and the two groups were assessed for reliability and validity. The intervention involved one 2-hour meeting every 2 weeks for a total of six meetings. The topics included goal setting, problem solving, coping with daily stress, seeking social support, and staying motivated. The sessions consisted of introductions that highlighted the topic, group discussions, identifying areas of difficulty with self-care behaviors after renal transplant, and developing a set of goals and strategies to overcome these problems. RESULTS: The empowerment group reported significant increases both in terms of level of empowerment (F = 5.29, p = .023) based on age and time interaction (F = 9.86, p < .001) and in terms of self-care behaviors (F = 7.15, p = .009). Moreover, these increases were significantly larger than the increases recorded by the comparison group. In addition, these increases were particularly large in the older empowerment-group participants with lower pretest scores for empowerment. CONCLUSIONS: Empowerment support may be critical to improve the empowerment and self-care behaviors of renal transplant patients. The results of this study may be applied to improve patient education and empowerment programs for renal transplant patients. Furthermore, these programs may be adjusted to take into consideration the learning preferences or needs of different age groups.

A convenient method for the synthesis of amine-terminated poly(ethylene oxide) and poly(epsilon-caprolactone).

A convenient synthetic route to prepare amine-terminated poly(ethylene oxide) (PEO) and poly(epsilon-caprolactone) (PCL) was described. The strategy involved two-step reactions, the condensation of hydroxyl-terminated PEO and PCL with N-benzyloxycarbonyl amino acid followed by the catalytic hydrogenation under mild conditions. NMR and GPC measurements indicated that the reactions proceeded nearly quantitatively. Amine-terminated PEO thus prepared was used to initiate the polymerization of alpha-(N(epsilon)-benzyloxycarbonyl-L-lysine) N-carboxy anhydride [lys(Z)-NCA], and the results confirmed that the reactivity of the amino group was high.