Identification of bioactive candidate compounds responsible for oxidative challenge from hydro-ethanolic extract of Moringa oleifera leaves.

Free radicals trigger chain reaction and inflict damage to the cells and its components, which in turn ultimately interrupts their biological activities. To prevent free radical damage, together with an endogenous antioxidant system, an exogenous supply of antioxidant components to the body in the form of functional food or nutritional diet helps undeniably. Research conducted by the Natl. Inst. of Health claimed that Moringa oleifera Lam possess the highest antioxidant content among various natural food sources based on an oxygen radical absorbent capacity assay. In this study, a 90% (ethanol:distilled water—90:10) gradient solvent was identified as one of the best gradient solvents for the effectual extraction of bioactive components from M. oleifera leaves. This finding was confirmed by various antioxidant assays, including radical scavenging activity (that is, 1, 1-diphenyl-2-picrylhydrazyl, H2O2, and NO radical scavenging assay) and total antioxidant capacity (that is, ferric reducing antioxidant power and molybdenum assay). High-performance liquid chromatography (HPLC) fingerprints of the 90% gradient extract visually showed few specific peaks, which on further analysis, using HPLC–DAD–ESI–MS, were identified as flavonoids and their derivatives. Despite commonly reported flavonoids, that is, kaempferol and quercetin, we report here for the 1st time the presence of multiflorin-B and apigenin in M. oleifera leaves. These findings might help researchers to further scrutinize this high activity exhibiting gradient extract and its bio-active candidates for fruitful clinical/translational investigations

Hepatoprotective nature of phytoextracts against hepatotoxin induced animal models: a review

Liver is the largest vital organ in human body and plays a major role in various metabolism and excretion of xenobiotics within the body. Liver dysfunction is a foremost health problem that challenges not only health care professionals but also the pharmaceutical industry as more than 900 drugs are implicated in case of liver injury. Hepatotoxicity is caused by various toxic substances and certain pharmaceutical drugs which produce liver injury such as-carbon tetrachloride, thioacetamide, high doses of acetaminophen, anti-tubercular drugs, few chemotherapeutic agents etc. The existing modern synthetic drugs to treat liver disorders in this condition also cause furthermore liver damage/complications. Therefore, many herbal drugs from natural origin have become increasingly popular and their use is wide spread. These herbal medicines have been used in the treatment of liver diseases for a long time because of their antioxidants properties and tissue protective nature. Extensive researches have been carried out on medicinal plants; however, only few herbal plants have attracted the interest of researchers, to evaluate them for better protective/therapeutic agents for hepatoprotective against chemical induced liver toxicity. Various preclinical investigations have proved that the efficacy of medicinally important plants in the treatment of liver diseases/disorders. Hence, this article provided valuable evidence to the knowledge of investigated medicinal plants; especially those medicinal plants are suitable for hepatoprotective and therapeutic nature against liver toxicity

Data S1: Raw data compilation

N-Acetyl-p-Aminophenol (APAP), also known as acetaminophen, is the most commonly used over-the counter analgesic and antipyretic medication. However, its overdose leads to both liver and kidney damage. APAP-induced toxicity is considered as one of the primary causes of acute liver failure; numerous scientific reports have focused majorly on APAP hepatotoxicity. Alternatively, not many works approach APAP nephrotoxicity focusing on both its mechanisms of action and therapeutic exploration. Moringa oleifera (MO) is pervasive in nature, is reported to possess a surplus amount of nutrients, and is enriched with several bioactive candidates including trace elements that act as curatives for various clinical conditions. In this study, we evaluated the nephro-protective potential of MO leaf extract against APAP nephrotoxicity in male Balb/c mice. A single-dose acute oral toxicity design was implemented in this study. Group 2, 3, 4 and 5 received a toxic dose of APAP (400 mg/kg of bw, i.p) and after an hour, these groups were administered with saline (10 mL/kg), silymarin—positive control (100 mg/kg of bw, i.p), MO leaf extract (100 mg/kg of bw, i.p), and MO leaf extract (200 mg/kg bw, i.p) respectively. Group 1 was administered saline (10 mL/kg) during both the sessions. APAP-treated mice exhibited a significant elevation of serum creatinine, blood urea nitrogen, sodium, potassium and chloride levels. A remarkable depletion of antioxidant enzymes such as SOD, CAT and GSH-Px with elevated MDA levels has been observed in APAP treated kidney tissues. They also exhibited a significant rise in pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) and decreased anti-inflammatory (IL-10) cytokine level in the kidney tissues. Disorganized glomerulus and dilated tubules with inflammatory cell infiltration were clearly observed in the histology of APAP treated mice kidneys. All these pathological changes were reversed in a dose-dependent manner after MO leaf extract treatment. Therefore, MO leaf extract has demonstrated some therapeutic effectiveness against APAP-induced nephrotoxicity through enhancement of the endogenous antioxidant system and a modulatory effect on specific inflammatory cytokines in kidney tissues

The modulatory effect of Moringa oleifera leaf extract on endogenous antioxidant systems and inflammatory markers in acetaminophen induced nephrotoxic mice model

N-Acetyl-p-Aminophenol (APAP), also known as acetaminophen, is the most commonly used over-the counter analgesic and antipyretic medication. However, its overdose leads to both liver and kidney damage. APAP-induced toxicity is considered as one of the primary causes of acute liver failure; numerous scientific reports have focused majorly on APAP hepatotoxicity. Alternatively, not many works approach APAP nephrotoxicity focusing on both its mechanisms of action and therapeutic exploration. Moringa oleifera (MO) is pervasive in nature, is reported to possess a surplus amount of nutrients, and is enriched with several bioactive candidates including trace elements that act as curatives for various clinical conditions. In this study, we evaluated the nephro-protective potential of MO leaf extract against APAP nephrotoxicity in male Balb/c mice. A single-dose acute oral toxicity design was implemented in this study. Group 2, 3, 4 and 5 received a toxic dose of APAP (400 mg/kg of bw, i.p) and after an hour, these groups were administered with saline (10 mL/kg), silymarin-positive control (100 mg/kg of bw, i.p), MO leaf extract (100 mg/kg of bw, i.p), and MO leaf extract (200 mg/kg bw, i.p) respectively. Group 1 was administered saline (10 mL/kg) during both the sessions. APAP-treated mice exhibited a significant elevation of serum creatinine, blood urea nitrogen, sodium, potassium and chloride levels. A remarkable depletion of antioxidant enzymes such as SOD, CAT and GSH-Px with elevated MDA levels has been observed in APAP treated kidney tissues. They also exhibited a significant rise in pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) and decreased anti-inflammatory (IL-10) cytokine level in the kidney tissues. Disorganized glomerulus and dilated tubules with inflammatory cell infiltration were clearly observed in the histology of APAP treated mice kidneys. All these pathological changes were reversed in a dose-dependent manner after MO leaf extract treatment. Therefore, MO leaf extract has demonstrated some therapeutic effectiveness against APAP-induced nephrotoxicity through enhancement of the endogenous antioxidant system and a modulatory effect on specific inflammatory cytokines in kidney tissues

Biocompatible polymers coated on carboxylated nanotubes functionalized with betulinic acid for effective drug delivery

Chemically functionalized carbon nanotubes are highly suitable and promising materials for potential biomedical applications like drug delivery due to their distinct physico-chemical characteristics and unique architecture. However, they are often associated with problems like insoluble in physiological environment and cytotoxicity issue due to impurities and catalyst residues contained in the nanotubes. On the other hand, surface coating agents play an essential role in preventing the nanoparticles from excessive agglomeration as well as providing good water dispersibility by replacing the hydrophobic surfaces of nanoparticles with hydrophilic moieties. Therefore, we have prepared four types of biopolymer-coated single walled carbon nanotubes systems functionalized with anticancer drug, betulinic acid in the presence of Tween 20, Tween 80, polyethylene glycol and chitosan as a comparative study. The Fourier transform infrared spectroscopy studies confirm the bonding of the coating molecules with the SWBA and these results were further supported by Raman spectroscopy. All chemically coated samples were found to release the drug in a slow, sustained and prolonged fashion compared to the uncoated ones, with the best fit to pseudo-second order kinetic model. The cytotoxic effects of the synthesized samples were evaluated in mouse embryonic fibroblast cells (3T3) at 24, 48 and 72 h. The in vitro results reveal that the cytotoxicity of the samples were dependent upon the drug release profiles as well as the chemical components of the surface coating agents. In general, the initial burst, drug release pattern and cytotoxicity could be well-controlled by carefully selecting the desired materials to suit different therapeutic applications

Graphene oxide as a nanocarrier for controlled release and targeted delivery of an anticancer active agent, chlorogenic acid

We have synthesized graphene oxide using improved Hummer's method in order to explore the potential use of the resulting graphene oxide as a nanocarrier for an active anticancer agent, chlorogenic acid (CA). The synthesized graphene oxide and chlorogenic acid-graphene oxide nanocomposite (CAGO) were characterized using Fourier transform infrared (FTIR) spectroscopy, thermogravimetry and differential thermogravimetry analysis, Raman spectroscopy, powder X-ray diffraction (PXRD), UV–vis spectroscopy and high resolution transmission electron microscopy (HRTEM) techniques. The successful conjugation of chlorogenic acid onto graphene oxide through hydrogen bonding and π–π interaction was confirmed by Raman spectroscopy, FTIR analysis and X-ray diffraction patterns. The loading of CA in the nanohybrid was estimated to be around 13.1% by UV–vis spectroscopy. The release profiles showed favourable, sustained and pH-dependent release of CA from CAGO nanocomposite and conformed well to the pseudo-second order kinetic model. Furthermore, the designed anticancer nanohybrid was thermally more stable than its counterpart. The in vitro cytotoxicity results revealed insignificant toxicity effect towards normal cell line, with a viability of > 80% even at higher concentration of 50 μg/mL. Contrarily, CAGO nanocomposite revealed enhanced toxic effect towards evaluated cancer cell lines (HepG2 human liver hepatocellular carcinoma cell line, A549 human lung adenocarcinoma epithelial cell line, and HeLa human cervical cancer cell line) compared to its free form

Characterization and formulation of bio-active fraction of Moringa oleifera Lam. leaves extract and its protective potential against acetaminophen toxicity

Moringa oleifera (MO) is a well-known and widely distributed tropical species of Moringaceae family. Its leaves possess an excellent nutritional profile and an impressive range of therapeutic properties. Recently, the investigations on pharmaceutical properties of MO leaves get expanded due to its enriched antioxidant potential. Though numerous study reports focused on its therapeutic efficacy, the responsible active compounds and its underlying molecular mechanism of action has not been determined yet which hold a setback for researchers to explore its exact therapeutic potential. Acetaminophen (APAP) overdose is a worldwide leading cause of acute liver failure and drug-induced hepatotoxicity. During APAP overdose, majority of the drug is converted by the cytochrome P450 (CYP 450 - 2E1/1A2) enzymes to the reactive toxic metabolite, N-acetyl-pbenzoquinoneimine (NAPQI) that depletes GSH level and covalently binds to the other cellular proteins and induce hepatocyte death/acute liver failure. Currently, the most effective therapy for APAP overdose is Nacetylcysteine (NAC), which replenishes glutathione level and enhances hepatic recovery. However, NAC has few significant limitations such as time constraints and reversal of GSH level alone may not be sufficient to arrest progress of APAP hepatotoxicity. This drives scientists/researchers in exploring for an alternative safe and effective therapy. In this study, the optimal MO gradient leaf extract has been obtained as 90% hydro-ethanolic solution based upon in vitro antioxidant assays and the active compounds responsible for its elite activity has been determined as quercetin, kaempferol, apigenin and multiflorin-B through chromatographic analysis. The underlying mechanism of action of 90% hydro-ethanolic MO leaf extract has been evaluated in Balb/c mice inflicted with lethal dose of APAP for hepato- and nephro-toxicity. The MO leaf extract effectively protects the liver through suppression of CYP 450 isoenzymes and in both liver and kidney through regulation of antioxidant enzymes level and modulation of inflammatory cytokines thereby hindering the further exacerbation of necrotic and renal tubular damage respectively. Further, 90% MO leaf crude extract was fractionated through liquid-liquid partition technique. Among the obtained solvent fractions, ethyl acetate (EA) fraction revealed the highest antioxidant activity evidently due to the presence of quercetin, kaempferol and apigenin which has been identified and quantified with commercial standards using HPLC analysis. Wherein, kaempferol was expressed in higher concentration with 263.86 μg, followed by apigenin and quercetin with 82.64 and 66.89 μg respectively, per mg of MO leaves EA fraction. Soy phosphatidylcholine (PC) is a bifunctional complex comprises of lipophilic phosphatidyl moiety and hydrophilic choline moiety. Naturally, flavonoids and phenolic compounds got affinity to bind with PC molecule resulting in a cell like lipid compatible molecular complex. In accordance, the EA fraction and its three major flavonoids quercetin, kaempferol and apigenin has been successfully loaded in PC molecule to retain its synergism and enhance bioavailability. Further its physico-chemical parameters, invitro drug release and hepato-protective potential against APAP inflicted hepatotoxicity in HepaRG cell line has been evaluated. The findings of this study has evidently suggested that MO leaves extract and its EA fraction loaded phospholipid complex can be implied as an effective antidote against APAP intoxication as it hinders/suppresses/modulates various key biomarkers involved in APAP hepatotoxicity pathway