Potent Protective Activity Research Articles

Discovery of a chalcone derivative as potent necroptosis inhibitor for the treatment of acute kidney injury.

Necroptosis, a form of inflammation-related programmed cell death, is a major mechanism of proximal tubular cell injury in acute kidney injury (AKI). Blockade of necroptosis signalling represents a promising strategy for clinical therapy of AKI. Previously, we identified a small molecular receptor-interacting protein kinases (RIPK)1 inhibitor Cpd-71 with nephroprotective activities. To discover more nephroprotective agents, in this study, 20 chalcone derivatives were synthesized and evaluated for their anti-necroptosis and nephroprotective activities. Among the chalcone derivatives, Cpd-2 exhibited the most potent anti-necroptosis activity (IC50 =1.08 μM) and protective activity (EC50 =1.49 μM) through directly binding to RIPK1 and blocking RIPK1-RIPK3-mixed-lineage kinase domain-like protein (MLKL) signalling pathway. Furthermore, Cpd-2 effectively attenuated cisplatin or hypoxia/reoxygenation (H/R)-induced injury and necroptotic inflammation in renal cell models. Moreover, in cisplatin- or ischemia/reperfusion (I/R) induced AKI mouse model, detection of creatinine and urea nitrogen in blood showed that Cpd-2 improved kidney function. Periodic acid-Schiff (PAS) staining and immunofluorescence analysis indicated that Cpd-2 also reduced pathological damage and inhibited inflammatory development in kidney tissues. In summary, although some chalcone derivatives have been reported to prevent kidney injury previously, our present study not only discovered a promising leading compound Cpd-2, but also provided a novel and successful practice for the development of necroptosis inhibitors from natural products derivatives as AKI therapeutic agents.

Development and biological evaluation of protective effect of kidney targeted N-acetylated chitosan nanoparticles containing thymoquinone for the treatment of DNA damage in cyclophosphamide-induced haemorrhagic cystitis

Thymoquinone (TQ), the most prominent constituent of Nigella sativa seeds, essential oil, is reported to possess an organ protective effect via Nrf2 expression and activation of Phase-II antioxidant enzymes. Haemorrhagic cystitis is the sudden onset of haematuria combined with bladder pain and irritable bladder symptoms are the known toxic effects of cyclophosphamide (CYP) chemotherapy. The objective of the present study was to investigate and compare the protective effect of thymoquinone (TQ) and thymoquinone nanoparticles (TQ-NP) in the kidney against CYP-induced haemorrhagic cystitis. Primarily, TQ-NP was fabricated by synthesis of N-acetylated chitosan and nanoparticle preparation by the ionic gelation technique. They were characterized by particle size, polydispersive index (PDI), zeta potential, entrapment efficiency (EE), SEM, and dynamic scattering calorimetry (DSC). Moreover, fluorescein isothiocyanate (FITC) labeled NPs were prepared for biodistribution studies. The protective mechanisms of TQ-NP included its anti-inflammatory activity, inhibitory effects on cytokine levels, and protection against the DNA damage in the bladder epithelium. The cystitis was induced in rats by orally administering 200 mg/kg of CYP. The dose-dependent protective effect of the TQ-NP was determined by intravenously administering 1, 2, and 5 mg/kg of the TQ-NP to CYP-treated rats. The present study revealed that the TQ-NP prepared by ionic gelation method provides kidney targeted delivery of TQ as compared to TQ solution. The mean particle size, PDI, and %EE of TQ-NP were 272.6 nm, 0.216, 70.81 ± 0.12% respectively. The zeta potential of thymoquinone-loaded nanoparticles was found to be −20.7 mV and − 22.6 mV respectively before and after lyophilization. SEM study also confirmed the small size and spherical shape. Pharmacokinetic studies revealed the improvement in half-life and prolonged action of the TQ-NP as compared to the TQ solution. Also, TQ-NP administration showed more protection against the characteristic histological alterations in the bladder in comparison to TQ solution. The present study indicates that TQ-NP exerts potent anti-oxidant, DNA protective and cytokine inhibitory activity at considerably lower concentrations as compared to plain TQ solution. The nano formulation of TQ using N-acetylated chitosan provides effective kidney targeted delivery of TQ, which in turn improves its retention and protective efficacy against CYP-induced haemorrhagic cystitis.

Discovery of 1,3,4-oxadiazole derivatives containing a bisamide moiety as a novel class of potential cardioprotective agents

Myocardial injury is a nonnegligible problem in cardiovascular diseases and cancer therapy. The functional feature of N-containing heterocycles in the cardiovascular field has attracted much attention in recent years. Herein, we discovered a lead compound 12a containing 1,3,4-oxadiazole by extensive screening of anticancer derivatives containing nitrogen-heterocycle, which exhibited potential protective activity against oxidative stress in cardiomyocytes. Follow-up structure-activity relationship (SAR) studies also highlighted the role of substitution sites and bisamide moiety in enhancing the protective activity against oxidative stress. Specifically, compound 12d exhibited low cytotoxicity under high concentration and potent myocardial protection against oxidative stress in H9c2 cells. Preliminary mechanistic studies showed compound 12d could decrease the expression of cardiac hypertrophy and oxidative stress-related proteins/genes and reduce mitochondria-mediated cell apoptosis, thereby enhancing the cell vitality of injured cardiomyocytes. In this study, 1,3,4-oxadiazole may represent a novel pharmacophore that possesses potential myocardial protection and provides more choices for future optimization of cardiovascular drugs, especially for the treatment of onco-cardiology.

Neurobehavioural and Histological Alterations in Lead Acetate-Exposed Rats Pretreated with Aqueous Leaf Extract of Vernonia amygdalina

Lead is a systemic toxicant that affects virtually every organ system, primarily the central nervous system, through an increase in the production of reactive oxygen species (ROS). This increase in ROS overwhelms the natural antioxidant system of the body leading to oxidative stress. Some plants such as Vernonia amygdalina have been reported to possess antioxidant activity which counteracts the damage induced by free radicals. Accordingly, this study was designed to investigate the possible protective activity of aqueous Vernonia amygdalina leaf extract against lead-induced neurotoxicity in adult Wistar rats. Thirty Wistar rats were randomised into six groups (A-F) consisting of five rats each for a study period of 28 days. Group A served as the control group, Group B was administered with 100mg/kg body weight of lead only, Groups C and D were pretreated with 200mg/kg and 400mg/kg bodyweight of aqueous Vernonia amygdalina leaf extract and 100mg/kg body weight of lead respectively. Groups E and F were administered with 200mg/kg and 400mg/kg body weight of aqueous Vernonia amygdalina leaf extract only. The Open field and Novel Object Recognition tests were assessed and thereafter, the cerebellum, cerebrum and hippocampus were harvested for histological examination. Results showed a significant decrease in the body and brain weights, ambulation, rearing and discrimination index as well as a conversely significant increase in grooming and immobility in lead-treated rats. Histological examination of lead-alone treated groups showed loss of some Purkinje cells in the cerebellum, degenerating pyramidal cells in the cerebrum and altered morphology with pyknotic nuclei in the hippocampus. Pretreatment of rats with Vernonia amygdalinaattenuated lead-induced alterations to body and brain weights, neurobehavioural activities and tissue histomorphology, thus indicating a potent protective activity. These findings provide the first research evidence that Vernonia amygdalina protects against lead-induced neurotoxicity in Wistar rats.

Exploration of Aqueous Phyllanthus amarus Leaf Extract as a Protective Agent in Mercury Chloride-Exposed Wistar Rats: A Neurobehavioural Study

Mercury (Hg) exposure is known to damage vital components of the central nervous system primarily through oxidative stress. Although Phyllanthus amarus (PA) is reported to be rich in antioxidants, there is a dearth of relevant research evidence to show its protective activity against Mercuric chloride (HgCl2)-induced neurotoxicity. Accordingly, this study was designed to investigate the possible protective activity of aqueous Phyllanthus amarus leaf extract (APALE) in HgCl2-exposed rats using standard neurobehavioral assessments. Thirty adult Wistar rats were randomized into six groups of five rats each viz: A: Control, distilled water; B: HgCl2 (4 mg/kg body wt.); C: APALE (200 mg/kg body wt.) + HgCl2 (4 mg/kg); D: APALE (400 mg/kg body wt.) + HgCl2 (4 mg/kg); E: APALE (200 mg/kg body wt.); F: APALE (400 mg/kg body wt.). All administrations were through an orogastric tube for 14 days. Thereafter, neurobehavioral tests (open field, novel object and Y-maze) were done and experimental rats were sacrificed by cervical dislocation. Body, brain and cerebellar weights were examined across all groups. HgCl2 exposure significantly (p<0.0.05) decreased whole body weight across experimental groups. Similarly, a reduction in brain and cerebellar weight was observed. Significant neurobehavioral alterations to locomotion, discrimination index and cognition/memory, induced by HgCl2, were observed across experimental groups. However, protection against the deleterious effects induced by HgCl2 was observed following pretreatment with APALE. Conclusively, these findings show that APALE demonstrated potent protective activity against HgCl2-induced neurotoxicity and may be useful in the management of Hg toxicity and other related disorders.

Preparation of deer oil powder and its effect on acute gastric mucosal injury in rats.

In this paper, deer oil was used as a raw material to prepare deer oil powder by microencapsulation technology to study the potential protective activity of deer oil powder on ethanol-induced acute gastric mucosal injury in rats. The results show that the best process for preparing deer oil powder is: the solids account for 25% of the system content, the wall material Whey Milk Protein Isolate-Maltodextrin ratio is 1:5, the Sodium Stearyl Lactate-Glycerides of Monostearate and Distearate compound emulsifier content is 0.8%, the ratio is 1:1, and the deer oil accounts for 30% of the solid content, and the spray drying inlet temperature is 180°C. Deer oil powder can reduce total stomach injury, gastric mucosal congestion area, and injury score index. Without affecting the blood lipid level, it can reduce the content of Myeloperoxidase, enhance the activity of Superoxide Dismutase and Glutathione Peroxidase, and has good antioxidant activity. Enhance the expression of defense factors Estradiol, Epidermal Growth Factor, and Somatostatin. At the same time, it reduces the levels of Interleukin-1β and Interleukin-6 pro-inflammatory cytokines in gastric tissue and enhances the expression of anti-inflammatory factors Interleukin-4 and Interleukin-10. These results indicate that the gastric protection mechanism of deer oil powder may be related to the enhancement of mucosal defense factors, inhibition of inflammation, and oxidative stress. PRACTICAL APPLICATIONS: This study screened the optimal formula for preparing microencapsulated deer oil powder and proved for the first time that deer oil powder has a strong gastroprotective effect on ethanol-induced acute gastric injury in rats. The gastric protective mechanism of deer oil powder is mainly to reduce oxidative stress, inflammatory cytokine accumulation and to increase the content of defense factors. Therefore, deer oil powder can be used as a new source of gastric treatment drugs.

Salidroside Ameliorates Ischemia-Induced Neuronal Injury through AMPK Dependent and Independent Pathways to Maintain Mitochondrial Quality Control.

Salidroside, an active ingredient in Rhodiola rosea, has potent protective activity against cerebral ischemia. However, the mechanisms underlying its pharmacological actions are poorly understood. In this study, we employed a mouse middle cerebral artery occlusion (MCAO) and cellular oxygen and glucose deprivation (OGD) models to test the hypothesis that salidroside may restore mitochondrial quality control in neurons by modulating the relevant signaling. The results indicated that salidroside mitigated almost 40% the ischemia-induced brain infarct volumes in mice and the OGD-decreased viability of neurons to ameliorate the mitochondrial functions. Furthermore, salidroside treatment alleviated the OGD- or ischemia-induced imbalance of mitochondrial fission and fusion, mitophagy and promoted mitochondrial biogenesis in neurons by attenuating the AMPK activity. Moreover, salidroside alleviated 50% the OGD-promoted mitochondrial calcium fluorescence intensity and 5% mitochondria-associated membrane (MAM) area by down-regulating GRP75 expression independent of the AMPK signaling. Finally, similar findings were achieved in primary mouse neurons. Collectively, these data indicate that salidroside effectively restores the mitochondria dynamics, facilitates mitochondrial biogenesis by attenuating the AMPK signaling, and maintains calcium homeostasis in neurons independent of the AMPK activity.

Protective effects of antioxidant vitamins (C plus E) against oxidative damage induced by the insecticide imidacloprid in male rats

IntroductionImidacloprid (IMC) is a synthetic neonicotinoid insecticide that is used in agriculture against harmful insects.AimThis study aimed to investigate the adverse effects of the IMC formulation on biochemical, hematological parameters, oxidative stress markers, pro-inflammatory cytokines, alpha-fetoprotein (AFP), and testosterone levels in male rats, and to determine the potential protective activity of vitamin C and E against IMC insecticides in rats.Material and methodsThe rats were divided into four groups. The first group was used as a control. The second, third, and fourth groups received vitamin C (200 mg/kg) plus vitamin E (200 mg/kg), IMC (45 mg/kg), and IMC plus vitamin C and E, respectively, for 28 days. At the end of treatment, blood and tissue samples were collected. Biochemical, hematological parameters, oxidative stress markers, pro-inflammatory cytokines, AFP, and testosterone were evaluated.ResultsThe results elucidated that the IMC significantly caused elevation in the serum liver and kidney functions, lipid profile, AFP, and pro-inflammatory marker levels, as well as liver and kidney oxidative stress markers. While the serum testosterone level and complete blood count were disturbed, besides that, the liver and kidney antioxidant enzymes activities were decreased.ConclusionsThe protective effect of vitamins C plus E against IMC toxicity in the rat have a beneficial role in combating the adverse effects of IMC.

Discovery of sterically-hindered phenol compounds with potent cytoprotective activities against ox-LDL–induced retinal pigment epithelial cell death as a potential pharmacotherapy

Late-stage dry age-related macular degeneration (AMD) or geographic atrophy (GA) is an irreversible blinding condition characterized by degeneration of retinal pigment epithelium (RPE) and the associated photoreceptors. Clinical and genetic evidence supports a role for dysfunctional lipid processing and accumulation of harmful oxidized lipids in the pathogenesis of GA. Using an oxidized low-density lipoprotein (ox-LDL)-induced RPE death assay, we screened and identified sterically-hindered phenol compounds with potent protective activities for RPE. The phenol-containing PPARγ agonist, troglitazone, protected against ox-LDL–induced RPE cell death, whereas other more potent PPARγ agonists did not protect RPE cells. Knockdown of PPARγ did not affect the protective activity of troglitazone in RPE, confirming the protective function is not due to the thiazolidine (TZD) group of troglitazone. Prototypical hindered phenol trolox and its analogs potently protected against ox-LDL–induced RPE cell death whereas potent antioxidants without the phenol group failed to protect RPE. Hindered phenols preserved lysosomal integrity against ox-LDL–induced damage and FITC-labeled trolox was localized to the lysosomes in RPE cells. Analogs of trolox inhibited reactive oxygen species (ROS) formation induced by ox-LDL uptake in a dose-dependent fashion and were effective at sub-micromolar concentrations. Treatment with trolox analog 2,2,5,7,8-pentamethyl-6-chromanol (PMC) significantly induced the expression of the lysosomal protein NPC-1 and reduced intracellular cholesterol level upon ox-LDL uptake. Our data indicate that the lysosomal-localized hindered phenols are uniquely potent in protecting the RPE against the toxic effects of ox-LDL, and may represent a novel pharmacotherapy to preserve the vision in patients with GA.

Effect of Aqueous Enzymatic Extraction of Deer Oil on Its Components and Its Protective Effect on Gastric Mucosa Injury.

In this study, deer suet fat was used as a raw material to study the effects of aqueous enzymatic extraction of deer oil on its components, followed by studies into the potential protective activity, and related molecular mechanisms of deer oil on ethanol-induced acute gastric mucosal injury in rats. The results show that aqueous enzymatic extraction of deer oil not only has a high extraction yield and has a small effect on the content of active ingredients. Deer oil can reduce total stomach injury. Without affecting the blood lipid level, it can reduce the oxidative stress, which is manifested by reducing the content of myeloperoxidase (MPO) and enhancing the activity level of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). It also enhances the expression of defense factors prostaglandin (E2), epidermal growth factor (EGF), and somatostatin (SS), it inhibits apoptosis evidenced by the enhanced of Bcl-2 and decreased expression of cleavage of caspase-3 and Bax. At the same time, it reduces inflammation, which is manifested by reducing the expression of IL-1β, interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) gastric tissue pro-inflammatory cytokines, and enhancing the expression of anti-inflammatory factors IL-4 and IL-10, and inhibiting the mitogen-activated protein kinase/nuclear factor kappa B (MAPK/NF-κB) signaling pathway in gastric tissue.

Potential antioxidant effects of Narcissus tazetta phenolic compounds against cadmium chloride-induced hepatotoxicity in Swiss albino mice.

Narcissus tazetta (Amaryllidaceae) is a medicinal plant widely used for cut flowers and potted ornamental plant in Tunisia flora. The current study evaluated the phenolic composition and antioxidant properties of its flower extracts and investigated its potential protective activity against cadmium chloride (CdCl2)-induced hepatotoxicity in mice. Mice were divided into six groups of six each: group 1, serving as negative controls, received by intraperitoneal way only distilled water; group 2 received by intraperitoneal way CdCl2 (0.16 mg/kg bw); groups 3 and 4 received CdCl2 at the same dose of group 2 and 100 or 200 mg/kg bw of Narcissus tazetta flower extracts via oral route; groups 5 and 6, serving as positive controls, received only Narcissus tazetta flower extracts. Polyphenolic compounds of the extract were analyzed by colorimetric and high-performance liquid chromatography-mass spectrometry (HPLC-MS) methods. Total antioxidant activity and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging potential of the extract were estimated using colorimetric method. Results indicated that ethanolic flower extract contained high levels of total phenolic and flavonoid along with a strong total antioxidant and DPPH free radical scavenging activities. HPLC-MS analysis identified eight phenolic compounds, including rutin, kaempferol glycosides, and chlorogenic acids. The extract also exhibited marked hepatoprotective effects against CdCl2 toxicity by reducing hepatic levels of malondialdehyde, advanced oxidation protein products, hydrogen peroxide, metallothioneins, and DNA degradation. Additionally, co-administration of Narcissus tazetta flower extracts lowered the plasma activities of transaminases, gamma glutamyl transpeptidase, and lactate dehydrogenase and increased hepatic levels of reduced glutathione, nonprotein thiols, vitamin C, and catalase activity. The hepatoprotective effects of the extract were demonstrated by histopathological improvement of liver disorders. The current study provided ethnopharmacological application of Narcissus tazetta flower extracts against CdCl2-induced oxidative stress, suggesting its chemoprevention role of its phenolic compounds as a natural antioxidant.

Effects of Moringa Oleifera Seeds Aqueous Extract on Type-II Diabetic Nephropathy in Adult Male Albino Rat

Abstract Background: Because of the potential antioxidant, anti-inflammatory, and immune-modulating properties of Moringa oleifera,there is an increased interest in its potential capacity in the prevention and control of diabetes mellitus and lipids peroxidation. Aim of Study: Evaluating the potential antidiabetic efficacy and protective activities of aqueous extract of Moringa oleifera seeds in alloxan-induced diabetic nephropathy. Material and Methods: This experimental study wasstarted in September and ended at November, 2020. It wasperformed at Medical Physiology Department, Al-Azhar Faculty of Medicine, Cairo. Fifty adult male albino rats of a local strain were distributed randomly into 5 equal groups: Normal control group, control group received aqueous extract of Moringa seeds (500ml/kg body weight orally for 10 weeks), diabetic control group received a single dose of alloxan (140mg/kg body weight intraperitoneally), diabetic group received aqueous extract of Moringa seeds (500ml/kg body weight orally 10 weeks), and diabetic group received Moringa seeds aqueous extract before and after induction of diabetes in the same pervious doses. Results: Alloxan-induced diabetes mellitus was associated with significant higher levels of serum glucose, glycosylated hemoglobin (HbA1c), and lowered level of insulin as compared to the control normal group. It was also associated with histopathological changes of renal tissues and impaired renal function in the form of significant elevations of serum K+, Na+, creatinine, urea and uric acid levels as compared to the control normal group. Also, there were significant disturbance of lipid peroxidation represented by MDA Level and significant decrease in SOD level as compared to the control normal group. Moringa oleifera seeds aqueous extract showed signif-icant lower levels of serum glucose, HbA1c and elevated level of insulin as compared with the diabetic rats. It was also associated with histological regeneration of damaged renal tissues and restoration of renal function in the form of signif-icant decrease of serum K+, Na+, creatinine, urea and uric acid levels as compared to the diabetic group. Also, there were significant amelioration of lipid peroxidation represented by significant decrease of MDA level, and significant increase of SOD level as compared to the control normal group. These results were prominent in pretreated group with Moringa seeds aqueous extract. Conclusion: Moringa oleifera improved glycemic and lipidemic disturbances as well as have potent protective and regenerative effects for the kidney.

Protective and curative mechanisms of echinochrome against 7, 12-Dimethylbenz[a]anthracene -induced renal toxicity in rats

Echinochrome (Ech) is one of the most important bioactive substance which is found in shells, spines, and eggs of the sea urchins. Aim: the present study was carried out to evaluate the curative and protective effects of Ech pigment against DMBA -induced renal toxicity in rats. Methods: Experimental rats were assigned into two main groups; protective group (treated with Ech for 14 days then administrated DMBA) and curative group (administrated DMBA then treated with Ech for 14 days). Each group is divided into 3 sub-groups; control, DMBA (15 mg/ kg body, weight orally), and Ech (1 mg/ kg body, weight orally). Results: The oral administration of Ech decreased the concentrations of urea, creatinine, uric acid, and MDA and increased GSH and CAT levels in both protective and curative groups. Moreover, histology of kidney tissue improved after the treatment with Ech. Conclusions: The results of the present study demonstrated the potential protective and curative activities of Ech against renal toxicity induced by DMBA through inhibiting the metabolism of DMBA and restoring the balance between ROS formation and internal antioxidant enzymes by its powerful antioxidant activity.

Cutaneous Malassezia: Commensal, Pathogen, or Protector?

The skin microbial community is a multifunctional ecosystem aiding prevention of infections from transient pathogens, maintenance of host immune homeostasis, and skin health. A better understanding of the complex milieu of microbe-microbe and host-microbe interactions will be required to define the ecosystem’s optimal function and enable rational design of microbiome targeted interventions. Malassezia, a fungal genus currently comprising 18 species and numerous functionally distinct strains, are lipid-dependent basidiomycetous yeasts and integral components of the skin microbiome. The high proportion of Malassezia in the skin microbiome makes understanding their role in healthy and diseased skin crucial to development of functional skin health knowledge and understanding of normal, healthy skin homeostasis. Over the last decade, new tools for Malassezia culture, detection, and genetic manipulation have revealed not only the ubiquity of Malassezia on skin but new pathogenic roles in seborrheic dermatitis, psoriasis, Crohn’s disease, and pancreatic ductal carcinoma. Application of these tools continues to peel back the layers of Malassezia/skin interactions, including clear examples of pathogenicity, commensalism, and potential protective or beneficial activities creating mutualism. Our increased understanding of host- and microbe-specific interactions should lead to identification of key factors that maintain skin in a state of healthy mutualism or, in turn, initiate pathogenic changes. These approaches are leading toward development of new therapeutic targets and treatment options. This review discusses recent developments that have expanded our understanding of Malassezia’s role in the skin microbiome, with a focus on its multiple roles in health and disease as commensal, pathogen, and protector.

Potential Protective Activities of Extracts of Phellinus linteus and the Altered Expressions of GSTM3 on Age-Related Cataract.

Age-related cataract (ARC) is one of the leading causes of visual impairment and blindness worldwide among the elderly. Here, we used sodium selenite-induced cataract mouse model, which shares with similarities with human senile cataract to investigate whether the extracts of Phellinus linteus (PLE) could have the potential protective effects of ARC or not. The mice pups were randomly divided into 4 treatment groups (n = 7): (1) normal saline on postpartum day 26; (2) Na selenite injected s.c on day 26; (3) Na selenite s.c on day 26+ gavaged PLE (40 mg/kg) on days 26–47; and (4) Na selenite s.c on day 26 + resveratrol on days 26–47. On day 47, encapsulated lenses and plasma were analyzed for the levels of glutathione (GSH), superoxide dismutase (SOD), and malondialdehyde (MDA), a marker of lipid peroxidation. Lens epithelial cells (LECs) were also analyzed for the mRNA and protein expressions of glutathione S-transferase Mu (GSTM3). We demonstrated that PLE could prevent selenite-induced oxidative stress and cataract formation in mice by higher GSH and SOD and lower MDA in LECs, plasma, and liver tissues and the increases in the mRNA and protein expressions of GSTM3 in LECs. Our data show the increasing oxidative stress in selenite-induced cataract mice. Our data reveal the benefits of PLE for preventive activity in selenite-induced cataract in mice and there is a good possibility that PLE could ameliorate human senile cataract.

Cholinesterase inhibitory potential of Dillenia suffruticosa chemical constituents and protective effect against Aβ−induced toxicity in transgenic Caenorhabditis elegans model

BackgroundThe Dillenia suffruticosa leaves have been used in African tribal herbal medicine to relieve inflammatory conditions and memory loss. However, little is known about the phytoconstituents and their potential against cholinergic dysfunction and β-amyloid accumulation related to neurodegenerative disorder. PurposeThree triterpenoids and three flavonoids were isolated and investigated for acetylcholinesterase and butyrylcholinesterase inhibitions, including molecular docking mechanisms underlying the effects. The effect of the bioactive triterpenoids to delay the β-amyloid (Aβ)-induced paralysis of transgenic Caenorhabditis elegans strain GMC101 was also studied. MethodologyThe extraction, fractionation, and isolation of the phytoconstituents from D. suffruticosa leaves followed the results of cholinesterase inhibitory activities determined using the modified Ellman's method. The in silico molecular docking simulation studies investigated the mode of interactions of the phytoconstituents on Torpedo californica acetylcholinesterase (TcAChE) and human butyrylcholinesterase (huBuChE) enzymes. The transgenic C. elegans strain GMC101 was tested with and without betulinic acid (1) or koetjapic acid (2) on their potential to delay paralysis. ResultsBetulinic acid (1) had the highest potency in acetylcholinesterase inhibition (AChEI) and butyrylcholinesterase inhibition (BuChEI), followed by koetjapic acid (2), whereas stigmasterol glycoside (3) displayed the least potency. Vitexin (4), a C-glycoside displayed a higher potency than tiliroside (5), the O-glycosidic flavonoid against BuChE and AChE. Kaempferol (6), compared to its aglycone (5), was about 4 and 1.4 times more potent on AChEI and BuChEI, respectively. The selectivity indices suggested that the isolated phytoconstituents were dual AChE-BuChE inhibitors, more selective against BuChE than AChE, except 5. The molecular docking studies supported that small molecule like 6 only required very low binding energy unlike its glycoside (5), to dock fully at the overcrowded catalytic sites of AChE. Bulkier ligands docked more easily at lesser crowded BuChE active sites instead. For the first time, 2 was reported to delay Aβ-induced paralysis of transgenic C. elegans strain GMC101 and produced significantly longer survival time (p < 0.001) than 1. ConclusionThe studies demonstrated that bulky phytoconstituents from D. suffruticosa are more likely inhibitors of BuChE than AChE. Koetjapic acid (2) and betulinic acid (1) may possess protective potential against neurodegenerative disorders, related to their anti-cholinesterase and β-amyloid mechanisms.Significance statement:The bulkier phytoconstituents of D. suffruticosa are mainly triterpenoids and flavonoid glycosides which may have potential protective activity against memory loss and cognitive decline related to Alzheimer's disease.

Prophylactic administration of a clinically safe low dose of the COVID-19 drug candidate Rejuveinix (RJX) effectively prevents fatal cytokine storm and mitigates inflammatory organ injury in a mouse model of sepsis

Severe viral sepsis of coronavirus disease 2019 (COVID-19) is associated with a Cytokine Release Syndrome (CRS) and a high case fatality rate due to the development of Acute Respiratory Distress Syndrome (ARDS) and multi-organ failure in high-risk COVID-19 patients, including cancer patients undergoing chemotherapy. Here, we demonstrate that our COVID-19 drug candidate Rejuveinix (RJX) exhibits potent protective anti-inflammatory activity in the LPS-GalN mouse model of fatal sepsis and multi-organ failure at a dose level >10x lower than its maximum tolerated dose (MTD) for human subjects. In BALB/c mice challenged with an otherwise invariably fatal dose of LPS-GalN, prophylactic administration of 0.7 mL/kg RJX (Human equivalent dose=0.057 mL/ kg), corresponding to 7.5% of its clinical Maximum Tolerated Dose (MTD), prevented the cytokine storm, mitigated the oxidative stress and inflammatory tissue injury in lungs, liver, and heart, and significantly improved the survival outcome. Furthermore, RJX increased the levels of antioxidant enzymes SOD, CAT, and GSH-Px, and reduced oxidative stress in the brain. These results indicate that RJX has clinical potential as a prophylactic anti-inflammatory agent against severe sepsis, including viral sepsis in COVID-19 patients.

Screening of the bioactive compounds in Amphora coffeaeformis extract and evaluating its protective effects against deltamethrin toxicity in rats.

Pyrethroids are synthetic chemicals similar to the pyrethrins, but more toxic to insects and mammals and persistent in the environment than pyrethrins. This study aimed to identify the bioactive compounds of Amphora coffeaeformis extract (ACE) and to determine their potential protective activity against deltamtherin (DEL) insecticide in rats. Six groups of male albino rats were treated for 4 weeks included the control group, ACE-treated group (772 mg/kg b.w.), DEL-exposed group (13.5 mg/kg b.w.), DEL plus ACE-treated group, and the groups treated with ACE for 14 days before or after DEL. At the end of treatment, blood and tissue samples were collected for biochemical assays. The GC-MS identified 18 compounds; most of them are fatty acid methyl ester, and the HPLC identified 8 polyphenols and significant amounts of vitamins A, C, B1, B2, B9, and E. The in vivo results revealed that DEL induced significant alterations in hematological and serum biochemical parameters, oxidative stress markers, proinflammatory cytokines, and NF-κB. ACE protects against DEL toxicity, and the protection was more pronounced in the groups treated with ACE plus DEL or ACE after DEL suggesting that ACE could be used for the prevention or the treatment of DEL toxicity. It could be concluded that ACE is a promising candidate for the production of bioactive compounds and should be considered in the pharmaceutical and food application.

Rejuveinix Shows a Favorable Clinical Safety Profile in Human Subjects and Exhibits Potent Preclinical Protective Activity in the Lipopolysaccharide-Galactosamine Mouse Model of Acute Respiratory Distress Syndrome and Multi-Organ Failure.

Background: New treatment platforms that can prevent acute respiratory distress syndrome (ARDS) or reduce its mortality rate in high-risk coronavirus disease 2019 (COVID-19) patients, such as those with an underlying cancer, are urgently needed. Rejuveinix (RJX) is an intravenous formulation of anti-oxidants and anti-inflammatory agents. Its active ingredients include ascorbic acid, cyanocobalamin, thiamine hydrochloride, riboflavin 5′ phosphate, niacinamide, pyridoxine hydrochloride, and calcium D-pantothenate. RJX is being developed as an anti-inflammatory and anti-oxidant treatment platform for patients with sepsis, including COVID-19 patients with viral sepsis and ARDS. Here, we report its clinical safety profile in a phase 1 clinical study (ClinicalTrials.gov Identifier: NCT03680105) and its potent protective activity in the lipopolysaccharide galactosamine (LPS-GalN) mouse model of ARDS. Methods: A phase 1, double-blind, placebo-controlled, randomized, two-part, ascending dose-escalation study was performed in participating 76 healthy volunteer human subjects in compliance with the ICH (E6) good clinical practice guidelines to evaluate the safety, tolerability, pharmacokinetics, and pharmacodynamics of RJX (Protocol No. RPI003; ClinicalTrials.gov Identifier: NCT03680105). The ability of RJX to prevent fatal shock, ARDS, and multi-organ failure was examined in the well-established LPS-GalN mouse model of sepsis and ARDS. Standard methods were employed for the statistical analysis of data in both studies. Findings: In the phase 1 clinical study, no participant developed serious adverse events (SAEs) or Grade 3-Grade 4 adverse events (AEs) or prematurely discontinued participation in the study. In the non-clinical study, RJX exhibited potent and dose-dependent protective activity, decreased the inflammatory cytokine responses (interleukin-6, tumor necrosis factor alpha, transforming growth factor beta), and improved survival in the LPS-GalN mouse model of sepsis and ARDS. Histopathological examinations showed that RJX attenuated the LPS-GalN induced acute lung injury (ALI) and pulmonary edema as well as liver damage. Conclusion: RJX showed a very favorable safety profile and tolerability in human subjects. It shows potential to favorably affect the clinical course of high-risk COVID-19 by preventing ARDS and its complications.

Potential Protective Activity of Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP38) on In Vivo Model of Heart Irradiation Induced Cardiotoxicity

Cardiac toxicity is one of the major adverse effects associated with thoracic irradiation (IR), which to certain extent, limits the survival benefit of radiotherapy. As yet, there exists no effective intervention strategy except for primary prevention and treatment as heart failure in general. Pituitary adenylate cyclase-activating polypeptide with 38 residues (PACAP38) has been found to potentially anti-inflammatory and anti-oxidative stress effects. Here, we aim to investigate protective activity of radiation-induced cardiotoxicity in mice with PACAP38. C57/BL6 mice (male, 6-8 weeks) were randomly assigned for control, IR and IR+PACAP38 treat groups, respectively. The IR was performed on 6-MV X-ray beam energy with 20 Gy/1Fx dose and 300 cGy/min dose rate, with SSD for 100 cm and radiation field for 1×1 cm, using a medical linear accelerator. The IR group received IR followed by 1 and 6 months of follow up. The treatment groups were given PACAP38 (10 μg/100 μl) intraperitoneally (i.p.) 2 h before IR and additional doses were given at 24 h and 48 h after IR. Control mice received equal volume of saline i.p. The animal echocardiography was examined at the 1 and 6 months after IR. 18F-FDG PET/CT was performed at the 6 months after IR. SUV was calculated by ROI on the irradiated and unirradiated fields. The animals were sacrificed and heart tissues were harvested accordingly. The pathological changes in cardiac left ventricular sections from wild-type mice with IR were significantly different between the saline- and PACAP38-treated IR mice, moderate damage was observed in IR exposed mice. The cardiac injury was characterized by cardiomyocyte degeneration, eosinophilic enhancement, cytoplasmic vacuolization, nuclear condensation and myocardial fiber contorted. PACAP38-treated IR mice showed significantly amelioration on myocardial morphology. At day 30 and 6 months, both diastolic and systolic functions of mice appeared no functional damage in all of the groups measured by the index of echocardiography. The expression of NFκB transcript was increased by IR (P<0.01, compared to control) and was blunted by PACAP38 treatment (P<0.05) in irradiated murine cardiac tissues using qRT-PCR. The expression of NRF2 mRNA was elevated by IR (P<0.01 compared to control) and was further enhanced by the PACAP38 treatment. Six months after a single dose of 20 Gy, the heart of mice showed high 18F-FDG uptake in the IR field. The average SUVs of control and IR groups were 0.709 ± 0.080 and 1.476 ± 0.225, respectively (P<0.001). Significant difference was shown on the reduction of 18F-FDG activity between IR and PACAP38+IR (0.725 ± 0.049, P<0.01) indicates that PACAP38 reversed cardiac injury and protected heart function. PACAP38 showed potential to protect against early cardiotoxicity in heart exposed to radiation in mice. Our findings support further evaluation of PACAP38 as a future candidate for cardioprotectant in cardiac injury associated with radiation.