Gene expression profiling reveals underlying molecular mechanism of hepatoprotective effect of Phyllanthus niruri on thioacetamide-induced hepatotoxicity in Sprague Dawley rats.

BACKGROUND: The liver plays an essential role in the body by regulating several important metabolic functions. Liver injury is associated with the distortion of these functions causing many health problems. Pharmaceutical drugs treat liver disorders but cause further damage to it. Hence, herbal drugs are used worldwide and are becoming increasingly popular. METHODS: The hepatoprotective activity of Phyllanthus niruri (PN) was evaluated against liver cirrhosis induced by thioacetamide (TAA) in male Sprague Dawley rats. Rats received intraperitoneal injections of thioacetamide (TAA, 200 mg/kg, b.w. three times weekly) for eight weeks. Daily treatments with plant extract (200 mg/kg) were administered orally for eight weeks. At the end of the study, hepatic damage was evaluated by monitoring transforming growth factor (TGFß), collagen α1 (Collα1), matrix metalloproteinase-2 (MMP2) and tissue inhibitor of matrix metalloproteinase-1 (TIMP1) gene expression by real-time PCR. Moreover, different chromatographic techniques including column chromatography, thin layer chromatography, and Ultra Performance Liquid Chromatography (UPLC) with Liquid Chromatography/Mass Spectrometry (LC/MS) were used to isolate the active constituents of the plant. RESULTS: The results revealed that treatment with PN significantly reduced the effect of thioacetamide toxicity and exhibited effective hepatoprotective activity. The mechanism of the hepatoprotective effect of PN is proposed to be by normalizing ROSs. Additionally, PN treatment regulated the expression of TGFß, Collα1, MMP2, and TIMP1 genes. In the active fraction of P. niruri, the isolated chemical constituents were 4-O-caffeoylquinic acid and quercetin 3-O-rhamnoside. CONCLUSIONS: The results of the present study indicate that PN ethanol extracts possess hepatoprotective activity that is most likely because of the isolated chemical constituents.

Protective Role of Phyllanthus niruri Extract against Thioacetamide-Induced Liver Cirrhosis in Rat Model.

A preclinical study was performed to determine if the extract from Phyllanthus niruri (PN) plays a protective role against liver cirrhosis induced by thioacetamide (TAA) in rats. Initially, acute toxicity was tested and the results showed that the extract was benign when applied to healthy rats. Next, the therapeutic effect of the extract was investigated using five groups of rats: control, TAA, silymarin, and PN high dose and low dose groups. Significant differences were observed between the TAA group and the other groups regarding body and liver weights, liver biochemical parameters, total antioxidant capacity, lipid peroxidation, and oxidative stress enzyme levels. Gross visualization indicated coarse granules on the surface of the hepatotoxic rats' livers, in contrast to the smoother surface in the livers of the silymarin and PN-treated rats. Histopathological analysis revealed necrosis, lymphocytes infiltration in the centrilobular region, and fibrous connective tissue proliferation in the livers of the hepatotoxic rats. But, the livers of the treated rats had comparatively minimal inflammation and normal lobular architecture. Silymarin and PN treatments effectively restored these measurements closer to their normal levels. Progression of liver cirrhosis induced by TAA in rats can be intervened using the PN extract and these effects are comparable to those of silymarin.

Cytoprotective effect of benzyl N'-(5-chloro-indol-3-yl-methylidene)-hydrazinecarbodithioate against ethanol-induced gastric mucosal injury in rats.

Indolic compounds have attracted a lot of attention due to their interesting biological properties. The present study was performed to evaluate the subacute toxicity and anti-ulcer activity of BClHC against ethanol-induced gastric ulcers. Experimental animal groups were orally pre-treated with different doses of BClHC (50, 100, 200 and 400 mg/kg) in 10% Tween 20 solution (vehicle). Blank and ulcer control groups were pre-treated with vehicle. The positive group was orally pretreated with 20 mg/kg omeprazole. After one hour, all groups received absolute ethanol (5 mL/kg) to generate gastric mucosal injury except the blank control group which was administered the vehicle solution. After an additional hour, all rats were sacrificed, and the ulcer areas of the gastric walls determined. Grossly, the ulcer control group exhibited severe mucosal injury, whereas pre-treatment with either derivative or omeprazole resulted in significant protection of gastric mucosal injury. Flattening of gastric mucosal folds was also observed in rats pretreated with BClHC. Histological studies of the gastric wall of ulcer control group revealed severe damage of gastric mucosa, along with edema and leucocytes infiltration of the submucosal layer compared to rats pre-treated with either BClHC or omeprazole where there were marked gastric protection along with reduction or absence of edema and leucocytes infiltration of the submucosal layer. Subacute toxicity study with a higher dose of derivative (5 g/kg) did not manifest any toxicological signs in rats. In conclusions, the present finding suggests that benzyl N'-(5-chloroindol-3-ylmethylidene)hydrazinecarbodithioate promotes ulcer protection as ascertained by the comparative decreases in ulcer areas, reduction of edema and leucocytes infiltration of the submucosal layer.

Acute toxicity evaluation, antibacterial, antioxidant and immunomodulatory effects of Melastoma malabathricum.

Melastoma malabathricum (MM) is a well-known plant in Malaysian traditional medicine, locally known as senduduk. Its ethanol and aqueous extracts have been used in the present investigation to study the immunomodulatory role on human peripheral blood mononuclear cell (PBMC), and the DPPH, ABTS and FRAP free radical scavenging activities were also measured. Total flavonoids and total phenolic contents were assayed and the antibacterial effect was tested against four species of bacteria; two Gram-positive (Staphylococcus aureus and Streptococcus agalactiae) and two Gram-negative (Escherichia coli and Klebsilla pneumonia). The tests were carried out using the disc diffusion, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) methods. Moreover, the acute toxicity was evaluated in vivo on the ethanol extract of MM to establish its safety when administered orally. In our results, both extracts of MM showed abilities to scavenge DPPH and ABTS free radicals, IC(50) values: (11.599 ± 0.84, 10.573 ± 0.58 µmol/L) and (62.657 ± 0.78, 63.939 ± 0.48 µmol/L) for ethanol and aqueous extracts respectively. Indeed the ethanol extract evidenced high phenolic content (384.33 ± 0.005 mg/g), flavonoids contents (85.8 ± 0.009 mg/g) and ferric reducing antioxidant power (33,590 ± 0.038 mmol/g), with high activity against S. aureus and S. agalactiae (11 ± 0.3 and 12 ± 0.6 mm inhibition zones). Likewise, the percentage of peripheral blood mononuclear cells (PBMC) viability was increased in response to MM, IC(50) values (1.781 ± 1.2 and 6.545 ± 0.93 µg/mL) for ethanol and aqueous extracts, respectively. In addition, our results showed that the MM extract is safe even at a high dose of 5,000 mg/kg and has no oral toxicity. These findings suggest the excellent medicinal bioactivity of MM and explain the popularity of this plant in the folk medicine as a remedy for different illnesses.

Assessment of in vitro antioxidant, antibacterial and immune activation potentials of aqueous and ethanol extracts of Phyllanthus niruri.

BACKGROUND: Recently much attention has been paid to biologically active plants because of their low production cost and fewer adverse effects compared with chemical drugs. In the present investigation the bioactivity of Phyllanthus niruri ethanol and aqueous extracts was evaluated in vitro. RESULTS: The ethanol extract of P. niruri showed a high level of flavonoid content (123.9 ± 0.002 mg g⁻¹), while the aqueous extract showed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH; IC50 6.85 ± 1.80 µmol L⁻¹) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS; 46.44 ± 0.53 µmol L⁻¹) free radical scavenging activities with high phenol content (376 ± 0.02 mg g⁻¹) and elevated levels of ferric reducing antioxidant power (FRAP; 23 883 ± 0.019 mmol g⁻¹) with excellent antibacterial activity against Staphylococcus aureus (20 mm inhibition zone) and Streptococcus agalactiae (12 mm inhibition zone), respectively, in addition to the best immune activation potential of human peripheral blood mononuclear cells (450.5%). CONCLUSIONS: It is clear from our results that both extracts of P. niruri has excellent bioactivity roles via elevated levels of antibacterial, antioxidant and percentage of peripheral blood mononuclear cell proliferation, which could lead to the development of medications for clinical use.

Synthesis, characterization, acetylcholinesterase inhibition, molecular modeling and antioxidant activities of some novel Schiff bases derived from 1-(2-ketoiminoethyl)piperazines.

Some novel Schiff bases derived from 1-(2-ketoiminoethyl)piperazines were synthesized and characterized by mass spectroscopy, FTIR, UV-Visible, 1H and 13C-NMR. The compounds were tested for inhibitory activities on human acetylcholinesterase (hAChE), antioxidant activities, acute oral toxicity and further studied by molecular modeling techniques. The study identified the compound (DHP) to have the highest activity among the series in hAChE inhibition and DPPH assay while the compound LP revealed the highest activity in the FRAP assay. The hAChE inhibitory activity of DHP is comparable with that of propidium, a known AChE inhibitor. This high activity of DHP was checked by molecular modeling which showed that DHP could not be considered as a bivalent ligand due to its incapability to occupy the esteratic site (ES) region of the 3D crystal structure of hAChE. The antioxidant study unveiled varying results in 1,1-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays. This indicates mechanistic variations of the compounds in the two assays. The potential therapeutic applications and safety of these compounds were suggested for use as human acetylcholinesterase inhibitors and antioxidants.

1H-Indole-3-carbaldehyde azine.

The molecule of the title compound, C(18)H(14)N(4), lies on a center of inversion such that there is one half-mol-ecule in the asymmetric unit. The N-N single bond adopts a trans configuration and the indole fused-ring system is nearly coplanar with the -CH=N-N=CH- fragment [dihedral angle = 9.8 (2)°]. Adjacent mol-ecules are linked by indole-azine N-H⋯N hydrogen bonds into a layer motif.

1H-Indole-3-carbaldehyde thio-semi-carbazone.

The mol-ecules of the title compound, C(10)H(10)N(4)S, are linked by N-H(indole)⋯S hydrogen bonds to form a linear hydrogen-bonded chain. There are two independent mol-ecules in the asymmetric unit.

Bis[1,5-bis-(1H-indol-3-ylmethyl-ene)thio-carbazonato-κN,S]nickel(II) dimethyl sulfoxide disolvate.

The Ni atom in the crystal structure of the centrosymmetric title compound, [Ni(C(19)H(15)N(6)S)(2)]·2C(2)H(6)OS, is N,S-chelated by the deprotonated Schiff bases in a square-planar geometry. The -CH=N-N=C(S)-NH-N=CH- frament is planar. The two indolyl -NH (donor) sites inter-act with dimethyl sulfoxide mol-ecules to furnish a layer motif.

trans-Bis(1H-indole-3-carbaldehyde thio-semicarbazonato-κN,S)nickel(II).

The Ni atom in the centrosymmetric title compound, [Ni(C(10)H(9)N(4)S)(2)], is N,S-chelated by the deprotonated Schiff bases in a square-planar geometry. The -CH=N-N=C(S)-NH(2) frament is planar. Adjacent mol-ecules are linked by hydrogen bonds between the indolyl -NH (donor) site and the double-bond =N- (acceptor) site of an adjacent mol-ecule, forming a layer motif.

Bis{µ-N'-[1-(5-bromo-2-oxidophen-yl)ethyl-idene]benzene-sulfono-hydrazidato}-κO,N':N;κN:O,N'-bis-[(dimethyl sulfoxide-κO)copper(II)].

In the title centrosymmetric dinuclear complex, [Cu(2)(C(15)H(11)BrN(2)O(3)S)(2)(C(2)H(6)OS)(2)], the Cu(II) ion is N,O-chelated by a dianionic ligand, monocoordinated by the sulfonamide N atom of a symmetry-related ligand and coordinated by an O atom from a dimethyl sulfoxide ligand, forming a distorted square-planar coordination geometry.

Bis{4-chloro-2-[2-(1H-indol-3-yl)ethyl-imino-meth-yl]phenolato-κN,O}zinc(II).

The Zn atom in the title compound, [Zn(C(17)H(14)ClN(2)O)(2)], is N,O-chelated by two deprotonated Schiff base monoanionic ligands in a tetra-hedral coordination geometry. The Zn atom lies on a special position of site symmetry 2.

N'-(2,5-Dihydroxy-benzyl-idene)benzene-sulfonohydrazide.

In the title compound, C(13)H(12)N(2)O(4)S, the dihedral angle between the two aromatic rings is 89.5 (1)°. In the crystal structure, mol-ecules are linked by O-H⋯O(hydr-oxy) and N-H⋯O(sulfon-yl) hydrogen bonds, forming a ribbon that propagates along the b axis; there is also an intra-molecular O-H⋯N hydrogen bond.

N'-[1-(2-Hydr-oxy-5-methyl-phen-yl)ethyl-idene]benzene-sulfonohydrazide.

The two independent mol-ecules in the asymmetric unit of the title compound, C(15)H(16)N(2)O(3)S, are each linked by an N-H⋯O(sulfon-yl) hydrogen bond into a linear chain that runs along the shortest axis of the triclinic unit cell. The hydr-oxy groups are engaged in intra-molecular hydrogen bonding and the amino N atom shows pyramidal coordination.

N'-(2-Hydr-oxy-5-nitro-benzyl-idene)-2-(1H-indol-3-yl)acetohydrazide.

The mol-ecule of the title compound, C(17)H(14)N(4)O(4), uses its amide -NH- group to form a hydrogen bond to the amido -C(=O)- group of an adjacent mol-ecule to furnish a linear chain structure. The hydr-oxy group forms an intra-molecular hydrogen bond; the indolyl -NH- unit does not engage in any strong hydrogen-bonding inter-actions.

Dichloridobis(2-{1-[2-(1H-indol-3-yl)ethyl-iminio]eth-yl}phenolate-κO)zinc(II)-2-{1-[2-(1H-indol-3-yl)ethyl-iminio]eth-yl}phenolate (1/2).

In the mononuclear complex mol-ecule of the title compond, [ZnCl(2)(C(18)H(18)N(2)O)(2)]·2C(18)H(18)N(2)O, the Zn atom, which lies on a twofold rotation axis, is coordinated by phenolate O atoms in a tetra-hedral coordination geometry. The coordinated Schiff base uses its indole NH donor site to form a hydrogen bond to the negatively charged phenolate O atom of the uncoordinated zwitterionic Schiff base. There is an intra-molecular N-H⋯O hydrogen bond in the coordinated and uncoordinated Schiff bases. The indole NH site of the uncoordinated Schiff base does not engage in a hydrogen-bond inter-action. The CH(2)-CH(2) group in the uncoordinated Schiff base is disordered equally over two positions.

2-[2-(1H-indol-3-yl)ethyl-iminiomethyl]-4-nitro-phenolate.

The title Schiff base, C(17)H(15)N(3)O(3), exists in the zwitterionic form with the phenol H atom transferred to the imine group. Adjacent zwitterions are linked into a linear chain running along the a axis by an indole-hydr-oxy N-H⋯O hydrogen bond [3.100 (2) Å].

N'-(5-Fluoro-2-oxo-2,3-dihydro-1H-indol-3-yl-idene)benzene-sulfono-hydrazide.

The mol-ecule of the title compound, C(14)H(10)FN(3)O(3)S, consists of an indole unit and a phenylsulfonyl unit that are disposed in an approximately trans orientation relative to the N-N single bond. Two mol-ecules are arranged about a center of inversion, forming a hydrazide-carbonyl N-H⋯O hydrogen-bonded dimer; the dimers are linked by an indole-sulfonyl N-H⋯O hydrogen bond into a ribbon.

Bis{2-[2-(1H-indol-3-yl)ethyl-imino-meth-yl]phenolato-κN,O}nickel(II) N,N-dimethyl-formamide disolvate.

The Ni atom in the title compound, [Ni(C(17)H(15)N(2)O)(2)]·2C(3)H(7)NO, lies on a twofold rotation axis. It is N,O-chelated by the deprotonated Schiff base 2-[2-(1H-indol-3-yl)ethyl-imino-meth-yl]phenolate ligand in a square-planar coordination environment. The mol-ecule is linked to a solvent mol-ecule by an indole-dimethyl-formamide N-H⋯O hydrogen bond.