Naringenin Research Articles

Fabrication and characterization of zein-sodium alginate complex nanoparticles as an effective naringenin delivery system: Physicochemical stability, solubility, antioxidant activity

Naringenin (NAR) is a native polyphenolic compound that has a variety of physiological benefits. However, the application of NAR in functional food fields is limited due to its high hydrophobicity and insufficient oral bioavailability. The objective of this study is to prepare naringenin-encapsulated zein-sodium alginate nanoparticles (NAR-Z/SA NPs) to improve the antioxidant effects of NAR by increasing the water solubility. The spherical NAR-Z/SA NPs possessed increased encapsulation efficiency (83.92 ± 0.32%) and loading capacity (18.24 ± 0.73%). The X-ray diffraction and differential scanning calorimetry results confirmed that NAR was in an amorphous state after being encapsulated. Fourier transform infrared spectroscopy revealed that hydrogen bonding, electrostatic interaction, and hydrophobic interaction drove the formation of the NAR-Z/SA NPs. Meanwhile, NAR-Z/SA NPs exhibited desirable pH (3.0–8.0), ionic (0–50 mM) and heating treatment (80°C) stability. Furthermore, NAR-Z/SA NPs had improved solubility and antioxidant activity compared with free NAR. In addition, the NAR-Z/SA NPs showed sustained release of NAR under simulated gastrointestinal conditions. These findings provide an effective choice for the broad application of NAR in food industries.

Ophthalmic Intervention of Naringenin Decreases Vascular Endothelial Growth Factor by Counteracting Oxidative Stress and Cellular Damage in In Vivo Zebrafish.

Diabetes Mellitus is a metabolic disease that leads to microvascular complications like Diabetic retinopathy (DR), a major cause of blindness worldwide. Current medications for DR are expensive and report multiple side effects; therefore, an alternative medication that alleviates the disease condition is required. An interventional approach targeting the vascular endothelial growth factor (VEGF) remains a treatment strategy for DR. Anti-VEGF medicines are being investigated as the main therapy for managing vision-threatening complications of DR, such as diabetic macular oedema. Therefore, this study investigated the effect of flavonoid naringenin (NG) from citrus fruits on inhibiting early DR in zebrafish. When exposed to 130 mM glucose, the zebrafish larvae developed a hyperglycaemic condition accompanied by oxidative stress, cellular damage, and lipid peroxidation. Similarly, when adult zebrafish were exposed to 4% Glucose, high glucose levels were observed in the ocular region and massive destruction in the retinal membrane. High glucose upregulated the expression of VEGF. In comparison, the co-exposure to NG inhibited oxidative stress and cellular damage and restored the glutathione levels in the ocular region of the zebrafish larvae. NG regressed the glucose levels and cellular damage along with an inhibition of macular degeneration in the retina of adult zebrafish and normalized the overexpression of VEGF as a promising strategy for treating DR. Therefore, intervention of NG could alleviate the domestication of alternative medicine in ophthalmic research.

In-situ formation of nanoparticles from drug-loaded 3D polymeric matrices

The in-situ formation of nanoparticles from polymer-based solid medicines, although previously described, has been overlooked despite its potential to interfere with oral drug bioavailability. Such polymeric pharmaceuticals are becoming increasingly common on the market and can become even more popular due to the dizzying advance of 3D printing medicines. Hence, this work aimed to study this phenomenon during the dissolution of 3D printed tablets produced with three different polymers, hydroxypropylmethylcellulose acetate succinate (HPMCAS), polyvinyl alcohol (PVA), and Eudragit RL PO® (EUD RL) combined with plasticizers and the model drug naringenin (NAR). The components' interaction, dissolution behavior, and characteristics of the formed particles were investigated employing thermal, spectroscopic, mechanical, and chromatographic assays. All the systems generated stable spherical-shaped particles throughout 24 h, encapsulating over 25% of NAR. Results suggest encapsulation efficiencies variations may depend on interactions between polymer-drug, drug-plasticizer, and polymer-plasticizer, which formed stable nanoparticles even in the drug absence, as observed with the HPMCAS and EUD RL formulations. Additionally, components solubility in the medium and previous formulation treatments are also a decisive factor for nanoparticle formation. In particular, the treatment provided by hot-melt extrusion and FDM 3D printing affected the dissolution efficiency enhancing the interaction between the components, reverberating on particle size and particle formation kinetics mainly for HPMCAS and EUD RL. In conclusion, the 3D printing process influences the in-situ formation of nanoparticles, which can directly affect oral drug bioavailability and needs to be monitored.

Silk fibroin peptide self-assembled nanofibers delivered naringenin to alleviate cisplatin-induced acute kidney injury by inhibiting mtDNA-cGAS-STING pathway.

Silk fibroin (SF) has excellent biocompatibility and biodegradability as a biomaterial. The purity and molecular weight distribution of silk fibroin peptide (SFP) make it more suitable for medical application. In this study, SFP nanofibers (molecular weight ∼30kD) were prepared through CaCl2/H2O/C2H5OH solution decomposition and dialysis, and adsorbed naringenin (NGN) to obtain SFP/NGN NFs. In vitro results showed that SFP/NGN NFs increased the antioxidant activity of NGN and protected HK-2cells from cisplatin-induced damage. In vivo results also showed that SFP/NGN NFs protected mice from cisplatin-induced acute kidney injury (AKI). The mechanism results showed that cisplatin induced mitochondrial damage, as well as increased mitophagy and mtDNA release, which activated the cGAS-STING pathway and induced the expression of inflammatory factors such as IL-6 and TNF-α. Interestingly, SFP/NGN NFs further activated mitophagy and inhibited mtDNA release and cGAS-STING pathway. Demonstrated that mitophagy-mtDNA-cGAS-STING signal axis was involved in the kidney protection mechanism of SFP/NGN NFs. In conclusion, our study confirmed that SFP/NGN NFs are candidates for protection of cisplatin-induced AKI, which is worthy of further study.

The Enhanced Skin Permeation of Flavonoids Via the Application of a Coamorphous in a Microemulsion Formulation

Improving the permeability and solubility of poorly water-soluble compounds is a major difficulty in skin permeation. In this study, we investigated whether using a pharmaceutical technique such as applying coamorphous to a microemulsion enhances the skin permeation of polyphenolic compounds. The melt-quenching technique created the coamorphous system between naringenin (NRG) and hesperetin (HPT), two polyphenolic compounds with poor water solubility. By creating a supersaturated state, the aqueous solution of coamorphous NRG/HPT demonstrated improved NRG and HPT skin permeation. However, as both compounds precipitated, the supersaturation ratio decreased. In contrast to crystal compounds, incorporating coamorphous material into microemulsions enabled the preparation of microemulsions in a wider formulation range. Additionally, compared to microemulsions with crystal compounds and an aqueous suspension of coamorphous, microemulsions with coamorphous NRG/HPT increased skin permeation of both compounds by more than four times. These results suggested that interactions between NRG and HPT are maintained in the microemulsion and enhance both compounds' skin permeation. An approach for improving the skin permeation of poorly water-soluble chemicals would be to apply a coamorphous system to a microemulsion.

The Potential Protective Role of Naringenin against Dasatinib-Induced Hepatotoxicity.

Dasatinib (DASA) is a novel tyrosine kinase inhibitor, approved for leukemia treatment. However, the long-term use of DASA induces several complications, especially liver damage. On the other hand, Naringenin (NGN) is a potent antioxidant and anti-inflammatory agent which is known to exert protective effects in several liver disease animal models. Yet, the effect of NGN on DASA-induced hepatotoxicity has not been examined. This study investigated the hepatoprotective effects of NGN against DASA-induced acute liver injury, using a mouse model. The mice were given NGN (50, 100, and 200 mg/kg po) or saline for 7 days, followed by DASA on the eighth day (25 mg/kg p.o.). DASA treatment alone was found to cause overexpression of proinflammatory cytokines, such as interleukin-10 (IL-10), tumor necrosis factor-alpha (TNF-α), and malonyl aldehyde (MDA), whereas attenuation of antioxidant genes including superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), and glutathione peroxidase (GPx). Interestingly, a pretreatment with NGN + DASA resulted in minimizing the proinflammatory mediators and restoring the levels of antioxidant genes. In addition, there was evidence of necro-inflammatory changes in histopathological findings in the liver samples after DASA administration which remarkably reduced with NGN + DASA. Thus, this study revealed that NGN could minimize the hepatotoxicity induced by DASA by providing anti-inflammatory and antioxidant protection.

Deep eutectic solvent-ultrasound assisted extraction as a green approach for enhanced extraction of naringenin from Searsia tripartita and retained their bioactivities.

Naringenin (NA) is a natural flavonoid used in the formulation of a wide range of pharmaceutical, fragrance, and cosmetic products. In this research, NA was extracted from Searsia tripartita using an environmentally friendly, high efficiency extraction method: an ultrasound-assisted extraction with deep eutectic solvents (UAE-DES). Six natural deep eutectic solvent systems were tested. Choline chloride was used as the hydrogen bond acceptor (HBA), and formic acid, ethylene glycol, lactic acid, urea, glycerol, and citric acid were used as hydrogen bond donors (HBD). Based on the results of single-factor experiments, response surface methodology using a Box-Behnken design was applied to determine the optimal conditions for UAE-DES. According to the results, the optimal NA extraction parameters were as follows: DES-1 consisted of choline chloride (HBA) and formic acid (HBD) in a mole ratio of 2:1, an extraction time of 10 min, an extraction temperature of 50°C, an ultrasonic amplitude of 75 W, and a solid-liquid ratio of 1/60 g/mL. Extracted NA was shown to inhibit the activity of different enzymes in vitro, including α-amylase, acetylcholinesterase, butyrylcholinesterase, tyrosinase, elastase, collagenase, and hyaluronidase. Thus, the UAE-DES technique produced high-efficiency NA extraction while retaining bioactivity, implying broad application potential, and making it worthy of consideration as a high-throughput green extraction method.

Naringenin-Capped Silver Nanoparticles Amalgamated Gel for the Treatment of Cutaneous Candidiasis.

The current research was aimed to synthesize a phytomolecule, naringenin (NRG)-mediated silver nanoparticles (NRG-SNPs) to study their antifungal potential against Candida albicans (C. albicans) and Candida glabrata (C. glabrata). The NRG-SNPs were synthesized by using NRG as a reducing agent. The synthesis of NRG-SNPs was confirmed by a color change and surface plasmon resonance (SPR) peak at 425nm. Furthermore, the NRG-SNPs were analyzed for size, PDI, and zeta potential, which were found to be 35 ± 0.21nm, 0.19 ± 0.03, and 17.73 ± 0.92mV, respectively. In silico results demonstrated that NRG had a strong affinity towards the sterol 14α-demethylase. The docking with ceramide revealed the skin permeation efficiency of the NRG-SNPs. Next, the NRG-SNPs were loaded into the topical dermal dosage form (NRG-SNPs-TDDF) by formulating a gel using Carbopol Ultrez 10 NF. The MIC50 of NRG solution and TSC-SNPs against C. albicans was found to be 50µg/mL and 4.8µg/mL, respectively, significantly (P < 0.05) higher than 0.3625µg/mL of NRG-SNPs-TDDF. Correspondingly, MIC50 results were calculated against C. glabrata and the results of NRG, TSC-SNPs, NRG-SNPs-TDDF, and miconazole nitrate were found to be 50µg/mL, 9.6µg/mL, 0.3625µg/mL, and 3-µg/mL, respectively. Interestingly, MIC50 of NRG-SNPs-TDDF was significantly (P < 0.05) lower than MIC50 of miconazole nitrate against C. glabrata. The FICI (fractional inhibitory concentration index) value against both the C. albicans and C. glabrata was found to be 0.016 and 0.011, respectively, which indicated the synergistic antifungal activity of NRG-SNPs-TDDF. Thus, NRG-SNPs-TDDF warrants further in depth in vivo study under a set of stringent parameters for translating in to a clinically viable antifungal product.

Naringenin Prevents Renal Injury in Experimental Hyperuricemia Through Suppressing Xanthine Oxidase, Inflammation, Apoptotic Pathway, DNA Damage, and Activating Antioxidant System.

Background/Purpose: This research was performed to determine the effect of naringenin (NAR) in experimental hyperuricemia (HU) induced by potassium oxonate (PO) on uric acid levels and xanthine oxidase (XO), inflammation, apoptotic pathway, DNA damage, and antioxidant system in kidney tissue. Study Design: Wistar Albino rats were categorized into four groups: (1) Control group, (2) PO group, (3) [PO+NAR] (2 weeks) group, and (4) PO (2 weeks)+NAR (2 weeks) group. Methods: The first group was not administered any drug. In group 2, PO was administered intraperitoneally 250 mg/kg/day for 2 weeks. In the third group, 100 mg/kg/day NAR was given intraperitoneally 1 hr after PO injection for 2 weeks. In the fourth group, PO was injected for the first 2 weeks, followed by NAR injection for the second 2 weeks. Serum uric acid levels, XO, nuclear factor-kappa B, tumor necrosis factor-alpha, interleukin-17, cytochrome c, 8-Hydroxydeoxyguanosine (8-OHdG), glutathione peroxidase (GPx), and caspase-3 levels in kidney were determined. Results: HU increased the levels of inflammatory and apoptotic parameters, XO, and 8-OHdG levels in kidney. Administration of NAR caused a decrease in these values and an increase in GPx levels. Conclusions: The results of the study show that NAR treatment reduces serum uric acid levels, and apoptosis, inflammation, and DNA damage; increases antioxidant activity in kidney in experimental HU.

Simultaneous estimation of neratinib and naringenin from pharmaceuticals using a stability-indicating liquid chromatographic method: Application to analytical quality by design concept and multi-tool method greenness assessment

A validated stability-indicating simultaneous liquid chromatographic method was developed to estimate neratinib (NRB) and naringenin (NRN). We used the principles of analytical quality by design (AQbD) to develop a robust analytical method. The ACN%, pH, and flow rate were the three method variables investigated and optimized using the Box-Behnken experimental design (BBD). The optimized chromatographic condition used a mobile phase of ACN: phosphate buffer pH 3.8 (70:30 v/v) flowing at 0.6 mL.min−1 on a 250 mm C-18 column. Diode array detection was performed at 277 nm. The method was robust, linear (r2 = 0.999), accurate and precise. The detection and quantitation limits were 0.275 and 1.0 µg.mL−1 for both drugs. Forced degradation revealed non-interference of degradation peaks with the analyte peaks and ensured method specificity. The LC estimation demonstrated higher solubility of NRB and NRN in Precirol ATO5 and Tween 80 as solid lipid and surfactant, respectively. In addition, the entrapment efficiencies were 78 % and 86 % for SLNs of NRB and NRN, respectively. After 24 h, the In-vitro release study indicated that 40 % of the drugs were released from the pure drug suspension and >95 % released from the SLN. Afterwards, the results from five different greenness assessment approach established the present method's green nature. Overall, a green, federally robust, and flexible liquid chromatographic method was developed and can be applied routinely to estimate NRB and NRN from the developed nano formulation.

Naringenin ameliorates Cyclophosphamide-induced nephrotoxicity in experimental model

Cyclophosphamide (CP) is widely described in the management of several nonneoplastic and neoplastic disorders. Renal damage is the most reported toxic effect of CP in clinical practice.Our study aimed to evaluate the effect of Naringenin (NG) in attenuating renal damage induced by CP in an experimental model.A total of 32 rats were divided into four groups (n = 8): negative control: rats fed on a basal diet, positive control: rats injected intraperitoneally with CP 50 mg/kg of body weight/day, NG 100: rats treated with NG 100 mg/kg/day body orally with concomitant administration of CP as described before, and NG 200: rats treated with NG 200 mg/kg/day body orally daily + CP. At the end of the experimental protocol (21 days), blood creatinine and urea levels were measured. The antioxidant activities and lipid peroxidation products were measured in the renal tissues as indicators of oxidative damage. Histopathological examination and immunohistochemistry staining were also performed on renal tissues.Coadministration of NG along with CP significantly (p < 0.001) improved the renal function and antioxidant capacities compared with positive control animals. Furthermore, histopathological, and immunological examination of renal tissue confirmed the protective effect of NG against CP-induced nephrotoxicity.The current study showed that NG has the potential to protect CP-induced renal damage, which may be beneficial for further studies and the design of NG analogs to be useful in clinical practice against CP-induced nephrotoxicity.

Naringenin mitigates reserpine-induced anxiety-like behavior, neurodegeneration, and oxidative stress in male rats.

Reserpine (Res) induces anxiety-like behaviors, orofacial dyskinesia, and neurodegeneration in animals, the pathophysiology of which has been related to oxidative stress. The purpose of this study was to investigate whether naringenin (NG) could prevent reserpine-induced anxiety-like behaviors, orofacial dyskinesia, and neurodegeneration in male rats. Twenty-eight male rats were distributed into different groups as follows: Control rats; vehicle rats, which received the vehicles (normal saline, orally; acetic acid, intraperitoneally); Res rats (1 mg/kg/day) every other day for 3 days; and Res + NG rats, which received NG (50 mg/kg, orally, pre-treatment for 7 days), followed by Res. Administration of Res significantly increased chewing frequency compared with the control group (P < 0.01) and NG reversed the effect of Res on this factor (P < 0.05). Res induced an anxiety-like behavior in rats in the plus maze, and pre-treatment with NG improved this behavior. In addition, Res significantly increased the level of oxidative stress markers and degenerated neurons in the striatum; NG was able to ameliorate these damages. The results of this study demonstrated that Res caused behavioral disorders and increased the levels of oxidative stress in male rats; the use of NG was effective in treating these disorders. Therefore, NG should be considered as a preventive agent for reserpine-induced brain damage in male rats.

Naringenin and proanthocyanidins pre-treatment decreases synthesis and activity of gelatinases induced by zoledronic acid in a dental implant surface in vitro model

ObjectiveTo assess the effects of pre-treatment with proanthocyanidins (PA) flavonoids, from grape seed extract, and synthetic naringenin (NA) on the synthesis of matrix metalloproteinases (MMPs) gelatinases and their tissue inhibitors (TIMPs), as well as the gelatinolytic activity of MMPs by human gingival fibroblasts (HGF) and osteoblasts (Ob) exposed to zoledronic acid (ZA) in a dental implant surface in vitro model. DesignThe highest non-cytotoxic concentrations of NA and PA were determined for HGF (10 μg/mL; defined by previous study) and Ob (0.5 μg/mL; defined by prestoBlue assay). Then, HFG and Ob were individually seeded onto titanium discs, and after 24 h, cells were pre-treated (or not) with NA or PA, followed (or not) by exposure to ZA. Next, MMP-2, MMP-9, TIMP-1, TIMP-2 synthesis (ELISA), and gelatinolytic activity (in situ zymography) was evaluated. Data were analyzed by one-way ANOVA and Tukey tests (α = 0.05). ResultsZA treatment increased the synthesis (p < 0.05) and activity of MMPs; flavonoids pre-treatment controlled ZA-induced gelatinolytic effects, down-regulating MMPs synthesis (p < 0.05) and activity by HGF and Ob. For HGF, NA and PA pre-treatment did not up-regulate TIMP synthesis after ZA exposure (p > 0.05); for Ob, TIMP-2 was up-regulated (p < 0.05) by flavonoids, followed by ZA. ConclusionsNA and PA pre-treatment provides interesting results in the modulation of ZA deleterious effects, down-regulating MMP-2 and MMP-9 synthesis and activity by HGF and Ob and up-regulating TIMP-2 by Ob.

Naringenin blocks hepatic cadmium accumulation and suppresses cadmium-induced hepatotoxicity via amelioration of oxidative inflammatory signaling and apoptosis in rats

Liver is one of the targets of cadmium (Cd) bioaccumulation for hepatic damage and pathologies via oxidative inflammation and apoptosis. The current study explored whether the citrus flavonoid naringenin (NAR) could prevent hepatic accumulation of Cd and Cd hepatotoxicity in a rat model. Rats in group 1 received normal saline; group 2 received NAR (50 mg/kg body weight); group 3 received CdCl2 (5 mg/kg body weight); group 4 received NAR + CdCl2, for four consecutive weeks. Assays related to markers of oxidative stress, inflammation, and apoptosis were carried out using liver homogenate. Blood and liver sample analyses revealed significant elevation of blood and hepatic Cd levels coupled with prominent increases in alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activities, whereas the albumin and total protein levels were decreased considerably. Hepatic superoxide dismutase (SOD), catalase (CAT), glutathione peroxide (GPx) activities diminished significantly compared to control followed by marked increases in malondialdehyde (MDA) levels, and dysregulation in caspase and cytokine (TNF-α, IL-6, IL-4, IL-10) levels. However, it was found that in the rats administered NAR + Cd, the levels of Cd, hepatic enzymes, MDA, TNF-α, IL-6, and caspases-3/-9 were prominently reduced compared to the Cd group. The hepatic SOD, CAT, GPx, IL-4, IL-10, albumin, and total protein were markedly elevated along with alleviated hepatic histopathological abrasions. Taken together therefore, NAR is a potential flavonoid for blocking hepatic Cd bioaccumulation and consequent inhibition of Cd-induced oxidative inflammation and apoptotic effects on the liver of rats.

Progress of research on the role of active ingredients of Citri Reticulatae Pericarpium in liver injury

Liver is a vital organ responsible for metabolizing and detoxifying both endogenous and exogenous substances in the body. However, it is susceptible to damage from chemical and natural toxins. The high incidence and mortality rates of liver disease and its associated complications impose a significant economic burden and survival pressure on patients and their families. Various liver diseases exist, including cholestasis, viral and non-viral hepatitis, fatty liver disease, drug-induced liver injury, alcoholic liver injury, and severe end-stage liver diseases such as cirrhosis, hepatocellular carcinoma (HCC), and cholangiocellular carcinoma (CCA). Recent research has shown that flavonoids found in Citri Reticulatae Pericarpium (CRP) have the potential to normalize blood glucose, cholesterol levels, and liver lipid levels. Additionally, these flavonoids exhibit anti-inflammatory properties, prevent oxidation and lipid peroxidation, and reduce liver toxicity, thereby preventing liver injury. Given these promising findings, it is essential to explore the potential of active components in CRP for developing new drugs to treat liver diseases. Recent studies have revealed that flavonoids, including hesperidin (HD), hesperetin (HT), naringenin (NIN), nobiletin (NOB), naringin (NRG), tangerine (TN), and erodcyol (ED), are the primary bioactive components in CRP. These flavonoids exhibit various therapeutic effects on liver injury, including anti-oxidative stress, anti-cytotoxicity, anti-inflammatory, anti-fibrosis, and anti-tumor mechanisms. In this review, we have summarized the research progress on the hepatoprotective effects of HD, HT, NIN, NOB, NRG, TN, ED and limonene (LIM), highlighting their underlying molecular mechanisms. Despite their promising effects, the current clinical application of these active ingredients in CRP has some limitations. Therefore, further studies are needed to explore the full potential of these flavonoids and develop new therapeutic strategies for liver diseases. For this review, we conducted a systematic search of three databases (ScienceNet, PubMed, and Science Direct) up to July 2022, using the search terms "CRP active ingredient," "liver injury," and "flavonoids." The search data followed the PRISMA standard. Our findings indicate that flavonoids found in CRP can effectively reduce drug-induced liver injury, alcoholic liver injury, and non-alcoholic liver injury. These therapeutic effects are mainly attributed to the ability of flavonoids to improve liver resistance to oxidative stress and inflammation while normalizing cholesterol and liver lipid levels by exhibiting anti-free radical and anti-lipid peroxidation properties. Our review provides new insights into the potential of active components in CRP for preventing and treating liver injury by regulating various molecular targets within different cell signaling pathways. This information can aid in the development of novel therapeutic strategies for liver disease.

Hepatoprotective and Neuroprotective Effects of Naringenin against Lead-Induced Oxidative Stress, Inflammation, and Apoptosis in Rats.

Naringenin (NRG) is one of the most important naturally occurring flavonoids, predominantly found in some edible fruits, such as citrus species and tomatoes. It has several biological activities, such as antioxidant, antitumor, antiviral, antibacterial, anti-inflammatory, antiadipogenic, and cardioprotective effects. The heavy metal lead is toxic and triggers oxidative stress, which causes toxicity in many organs, including the liver and brain. This study explored the potential protective role of NRG in hepato- and neurotoxicity caused by lead acetate in rats. Four groups of ten male albino rats were included: group 1 was a control, group 2 was orally treated with lead acetate (LA) at a dose of 500 mg/kg BW, group 3 was treated with naringenin (NRG) at a dose of 50 mg/kg BW, and group 4 was treated with 500 mg/kg LA and 50 mg/kg NRG for 4 weeks. Then, blood was taken, the rats were euthanized, and liver and brain tissues were collected. The findings revealed that LA exposure induced hepatotoxicity with a significant increase in liver function markers (p < 0.05). In addition, albumin and total protein (TP) and the albumin/globulin ratio (A/G ratio) (p < 0.05) were markedly lowered, whereas the serum globulin level (p > 0.05) was unaltered. LA also induced oxidative damage, demonstrated by a significant increase in malonaldehyde (MDA) (p < 0.05), together with a pronounced antioxidant system reduction (SOD, CAT, and GSH) (p < 0.05) in both liver and brain tissues. Inflammation of the liver and brain caused by LA was indicated by increased levels of nuclear factor kappa beta (NF-κβ) and caspase-3, (p < 0.05), and the levels of B-cell lymphocyte-2 (BCL-2) and interleukin-10 (IL-10) (p < 0.05) were decreased. Brain tissue damage induced by LA toxicity was demonstrated by the downregulation of the neurotransmitters norepinephrine (NE), dopamine (DA), serotonin (5-HT), and creatine kinase (CK-BB) (p < 0.05). Additionally, the liver and brain of LA-treated rats displayed notable histopathological damage. In conclusion, NRG has potential hepato- and neuroprotective effects against lead acetate toxicity. However, additional research is needed in order to propose naringenin as a potential protective agent against renal and cardiac toxicity mediated by lead acetate.

Naringenin facilitates M2 macrophage polarization after myocardial ischemia-reperfusion by promoting nuclear translocation of transcription factor EBand inhibiting the NLRP3 inflammasome pathway.

Myocardial ischemia-reperfusion injury (MIRI) remains an unsolved puzzle in medical circles. Naringenin (NAR) is a flavonoid with cardioprotective potential. The purpose of this article was to discuss the protective mechanism of NAR in MIRI by regulating macrophage polarization. The MIRI mouse model was established and perfused with NAR before surgery. In the in vitro experiment, macrophages RAW264.7 were treated with lipopolysaccharide to induce M1 polarization after pretreatment with NAR. Rescue experiments were carried out to validate the functions of transcription factor EB (TFEB), the NLR pyrin domain containing 3 (NLRP3) inflammasome, and autophagy in macrophage polarization. NAR reduced histopathological injury and infarction of myocardial tissues in MIRI mice, inhibited M1 polarization and promoted M2 polarization of macrophages, diminished levels of pro-inflammatory factors, and augmented levels of anti-inflammatory factors. NAR facilitated TFEB nuclear translocation and inhibited the NLRP3 inflammasome pathway. Silencing TFEB or Nigericin partly nullified the effect of NAR on macrophage polarization. NAR increased autophagosome formation, autophagy flux, and autophagy level. Autophagy inhibitor 3-methyladenine partly invalidated the inhibition of NAR on the NLRP3 inflammasome pathway. In animal experiments, NAR protected MIRI mice through the TFEB-autophagy-NLRP3 inflammasome pathway. Collectively, NAR inhibited NLRP3 inflammasome activation and facilitated M2 macrophage polarization by stimulating TFEB nuclear translocation, thus protecting against MIRI.

Naringenin Attenuates Isoprenaline-Induced Cardiac Hypertrophy by Suppressing Oxidative Stress through the AMPK/NOX2/MAPK Signaling Pathway

Cardiac hypertrophy is accompanied by increased myocardial oxidative stress, and whether naringenin, a natural antioxidant, is effective in the therapy of cardiac hypertrophy remains unknown. In the present study, different dosage regimens (25, 50, and 100 mg/kg/d for three weeks) of naringenin (NAR) were orally gavaged in an isoprenaline (ISO) (7.5mg/kg)-induced cardiac hypertrophic C57BL/6J mouse model. The administration of ISO led to significant cardiac hypertrophy, which was alleviated by pretreatment with naringenin in both in vivo and in vitro experiments. Naringenin inhibited ISO-induced oxidative stress, as demonstrated by the increased SOD activity, decreased MDA level and NOX2 expression, and inhibited MAPK signaling. Meanwhile, after the pretreatment with compound C (a selective AMPK inhibitor), the anti-hypertrophic and anti-oxidative stress effects of naringenin were blocked, suggesting the protective effect of naringenin on cardiac hypertrophy. Our present study indicated that naringenin attenuated ISO-induced cardiac hypertrophy by regulating the AMPK/NOX2/MAPK signaling pathway.

New Insights into Traditional Chinese Medicine in Treatment of Diabetic Nephropathy

High-calorie diet and sedentary lifestyles are strongly associated with the development and onset of type 2 diabetes (T2D). According to the International Diabetes Federation, the global incidence of diabetes among individuals aged 20–79 years was 9.3% in 2019. The current estimated total number of patients with diabetes is 463 million, with an expected increase to over 578 million by 2030.[1] Systemic insulin resistance and insufficient insulin secretion in T2D result in several complications that severely impair a patients quality of life, especially the development of T2D nephropathy (T2DN).[2] Increasing evidence has shown that the dysregulation of TGF-β signaling contributes to the development of T2DN. For example, the TGF-β/Smad3 signaling pathway is dysregulated in the kidneys of T2D patients, where activation of Smad3 and loss of its inhibitor Smad7 lead to severe glomerular and tubulointerstitial fibrosis.[3] T2D associated with high levels of blood glucose and advanced glycation end-products can induce Smad3 phosphorylation in mesangial cells, tubular epithelial cells, and vascular smooth muscle cells in vitro.[3–4] Smad3 also reportedly downregulated the expression of the transcription factors Pdx-1, Nkx6.1, and NeuroD1 in the islet beta cells of db/db mice,[6,7] which transcriptionally suppresses the proliferation and function of islet beta cells and therefore further promotes the progression of T2D. In contrast, Smad7, a natural negative feedback regulator of Smad3, can reduce renal inflammation in diabetic nephropathy by promoting islet beta cell proliferation by targeting an NF-κB-dependent mechanism.[8] Thus, rebalancing the equilibrium between Smad3 and Smad7 in TGF-β downstream signaling may represent a therapeutic strategy for T2DN.[9] Traditional Chinese medicine (TCM) has been reported to have therapeutic effects in T2D. For example, a clinical study showed that Tianqi can significantly slow down diabetes progression and minimize diabetes risk by enhancing glucose tolerance.[10] Berberine, the main compound of Coptis, reduced insulin resistance in rats with streptozotocin-induced diabetes by suppressing oxidative stress and aldose reductase.[11] Combination therapy is a well-documented therapeutic protocol for TCM, which involves the simultaneous or sequential use of more than one herbal component to maximize the therapeutic outcome by parallel regulation of numerous signaling pathways and transcriptional events. In fact, our group has invented a novel TCM-based combination therapy named AANG, containing a natural Smad7 agonist, asiatic acid (AA) from Centella asiatica,[12] and the Smad3 inhibitor naringenin (NG) from Citrus or Millettiareticulata Benth,[13] which synergistically inhibit multidrug resistance in hepatocellular carcinoma by targeting Smad3-dependent p-glycoprotein expression at the transcriptional level.[14] These findings revealed the therapeutic potential of AANG in other human diseases associated with the aberration of TGF-β/Smad3 signaling. Encouragingly, our recently published article in the International Journal of Biological Sciences[15] provides novel insights into Smad3-dependent T2DN. In this study, we used AANG to rebalance the Smad3/Smad7 equilibrium in the diabetic injured kidneys of db/db mice, which not only reduced NF-κB-driven renal inflammation but also prevented Smad3-mediated renal fibrosis. Moreover, the collective results of the study found that AANG combination had a better effect than monotherapy with either AA or NG on the regulation of Smad3/Smad7 signaling in diabetic kidneys. Our data suggested that AANG effectively protected islet beta cells against diabetes-induced regression in a Smad3-dependent manner. Furthermore, we observed the synergistic effect of AANG on the induction of Smad7 for protecting the diabetic kidney against NF-κB-mediated renal inflammation.[16,17] In addition, we detected that AANG can inhibit the expression of the Smad3-dependent fibrogenic lncRNA Erbb4-IR[18] and inflammatory lncRNA LRNA9884[19] in db/db mice, which may also represent a protective mechanism against T2DN. Importantly, in the same study,[15] we unexpectedly discovered the preventive effect of AANG therapy on the primary disease T2D. We found that AANG synergistically prevented the onset of T2D in db/db mice, resulting in improved insulin sensitivity and glucose tolerance. The preventive effect of AANG on T2D was achieved by starting treatment in 4-week-old db/db mice at the pre-diabetic stage but not at the post-diabetic stage (12-week-old db/db mice). We further identified that AANG exerts a preventive effect on pancreatic islets, the niche of T2D onset. These interesting findings suggested that AANG can prevent T2D if applied to individuals at the pre-diabetic stage. In summary, our new study[15] demonstrated the efficiency of AANG in suppressing the development of T2DN in db/db mice by systematically rebalancing Smad3/Smad7 signaling in the pancreas and kidneys, which consequently suppressed Smad3-mediated renal fibrosis and NF-κB-driven renal inflammation. In addition, we unexpectedly discovered a novel therapeutic function of AANG in preventing T2D onset by reserving beta cell proliferation and insulin production capacity in islets under diabetic conditions. This research provides new insights and important clinical rationale for translating this AANG combined therapy as a unique and an effective treatment for T2DN, as well as a TCM-based formula for preventing T2D onset in patients with pre-diabetic conditions.

ANTI-FIBROTIC AND ANTIOXIDANT AMELIORATIVE EFFECTS OF NARINGENIN AGAINST THIOACETAMIDE INDUCED LIVER FIBROSIS

Liver fibrosis still life-threatening problem and searching out ameliorative products motives many scientists, so this work evaluated the anti-fibrotic and antioxidant role of naringenin (NAR) against thioacetamide (TAA) induced liver fibrosis. Fifty adult male albino rats randomly divided into 5 groups (10 each); the first kept as control; the second treated I/P by 200 mg/kg TAA twice a week for 8 weeks; the third was gavaged daily with 50 mg /kg/ b.wt of NAR for 8 weeks; the fourth was co-treated by TAA and NAR while the fifth was treated with TAA for 8 w then gavaged daily by NAR for 1 month. TAA administration significantly increases the hepatic cell enzymes (ALT, AST, ALP and GGT) in the serum referring to hepatic cell destruction with an increase in hepatic MDA with a reduction in GSH concentrations, antioxidant enzyme activities as well as down regulation of their expression levels. NAR administration either with or after TAA ameliorates this effect suggesting its antioxidant ability. In the fibrotic pathway, TAA treatment up-regulates the expression levels of fibrogenic biomarkers (TGF-β, collagen 1α and fibronectin) genes while NAR down-regulates these genes suggesting its anti-fibrotic ability. Histopathological analysis confirms the biochemical results. In conclusion, NAR ameliorates the deleterious effect of TAA through its antioxidant and anti-fibrotic abilities.