Investigation of Antidiabetic, Antihyperlipidemic, and In Vivo Antioxidant Properties of Sphaeranthus indicus Linn. in Type 1 Diabetic Rats: An Identification of Possible Biomarkers

The present investigation was aimed to study the antidiabetic, antihyperlipidemic, and in vivo antioxidant properties of the root of Sphaeranthus indicus Linn. in streptozotocin- (STZ-) induced type 1 diabetic rats. Administration of ethanolic extract of Sphaeranthus indicus root (EESIR) 100 and 200 mg/kg to the STZ-induced diabetic rats showed significant (P < .01) reduction in blood glucose and increase in body weight compared to diabetic control rats. Both the doses of EESIR-treated diabetic rats showed significant (P < .01) alteration in elevated lipid profile levels than diabetic control rats. The EESIR treatment in diabetic rats produced significant increase in superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and decrease in thiobarbituric acid reactive substances (TBARS) levels than diabetic control rats. Administration of EESIR 200 mg/kg produced significant (P < .01) higher antioxidant activity than EESIR 100 mg/kg. The high performance liquid chromatography (HPLC) analysis of EESIR revealed the presence of biomarkers gallic acid and quercetin. In conclusion, EESIR possess antidiabetic, antihyperlipidemic, and in vivo antioxidant activity in type 1 diabetic rats. Its antioxidant and lipid lowering effect will help to prevent diabetic complications, and these actions are possibly due to presence of above biomarkers

Evaluation of synergistic antimicrobial activity of Cinnamonum zeylanicum, Trachyspermum amni and Syzygium aromaticum

Abstract Antimicrobial assay of acetone and ethanol extract of Cinnamomum zeylancium, Trachyspermum ammi and Syzygium aromaticum was performed using agar well diffusion method against bacterial culture. (E.coli, P.mirabilis and K.pneumoniae) the acetone extract of Cinnamomum zeylancium, ethanol extract of Trachyspermum ammi and acetone extract of Syzygium aromaticum were selected to evaluate the synergistic activity. The activities were combined in the ratio of 1:1:1, 1:2:1 and 1:1:2 (Trachyspermum ammi: Cinnamonum zeylancum: Syzygium aromaticum). Phytochemical analysis was carried out for the ethanol and acetone extract of Cinnamonum zeylancium, Trachyspermum ammi and Syzygium aromaticum, to check the present of carbohydrate, proteins, sterioids, resins, tannins, glycosides, flavonoids, saponins and quinines

Non-Selective Cannabinoid Receptor Antagonists, Hinokiresinols Reduce Infiltration of Microglia/Macrophages into Ischemic Brain Lesions in Rat via Modulating 2-Arachidonolyglycerol-Induced Migration and Mitochondrial Activity

<div><p>Growing evidence suggests that therapeutic strategies to modulate the post-ischemic inflammatory responses are promising approaches to improve stroke outcome. Although the endocannabinoid system has been emerged as an endogenous therapeutic target to regulate inflammation after stroke insult, the downstream mechanisms and their potentials for therapeutic intervention remain controversial. Here we identified <i>trans</i>- and <i>cis</i>-hinokiresinols as novel non-selective antagonists for two G-protein-coupled cannabinoid receptors, cannabinoid receptor type 1 and type 2. The Electric Cell-substrate Impedance Sensing and Boyden chamber migration assays using primary microglial cultures revealed that both hinokiresinols significantly inhibited an endocannabinoid, 2-arachidonoylglycerol-induced migration. Hinokiresinols modulated 2-arachidonoylglycerol-induced mitochondrial bioenergetics in microglia as evidenced by inhibition of ATP turnover and reduction in respiratory capacity, thereby resulting in impaired migration activity. In rats subjected to transient middle cerebral artery occlusion (1.5-h) followed by 24-h reperfusion, post-ischemic treatment with hinokiresinols (2 and 7-h after the onset of ischemia, 10 mg/kg) significantly reduced cerebral infarct and infiltration of ED1-positive microglial/macrophage cells into cerebral ischemic lesions <i>in vivo</i>. Co-administration of exogenous 2-AG (1 mg/kg, i.v., single dose at 2 h after starting MCAO) abolished the protective effect of <i>trans</i>-hinokiresionol. These results suggest that hinokiresinols may serve as stroke treatment by targeting the endocannabinoid system. Alteration of mitochondrial bioenergetics and consequent inhibition of inflammatory cells migration may be a novel mechanism underlying anti-ischemic effects conferred by cannabinoid receptor antagonists.</p></div

Post-treatment of hinokiresinols significantly reduced ED-1-postive microglia/macrophages in ischemic lesions.

<p>Rats were subject to MCAO for 90 min and then reperfused. After 24 h, coronal brain sections were stained with triphenyltetrazolium chloride (TTC) for brain infarct, anti-ED-1 antibody to detect macrophages/monocytes/activated microglia (red), or Hoechst for nuclei staining (blue). Data are presented as mean ± SEM. Scale bar = 50 μm. ^*<i>P</i> < 0.05: vs. indicated group. (A) <i>Trans-</i> or <i>cis</i>-hinokiresinol (10 mg/kg, i.p.) was administrated at 2 h and 7 h after the onset of MCAO. Representative images (left panel) and quantification of cell counts of ED-1-positive cells (right panel) are provided. N = 3–5. (B) Co-administration of 2-AG abolished the protective effect of <i>trans</i>-hinokiresionol. <i>Trans-</i> hinokiresinol was administrated in the presence or absence of 10 mg/kg 2-AG (once at 2 h after starting MCAO, i.v.). Representative images (left panel) and quantification (right) of either cell counts of ED-1-positive cells (upper) or infarct volume (lower) are provided. N = 4–7.</p

Hinokiresinols reduces 2-AG-induced microglial migration.

<p>(A) Schematic configuration of ECIS/taxis electrode design to measure an increase of resistance proportional to cell migration (left) and representative trace of ECIS change in the presence of hinokiresinols (right). The chemotactic response of pure primary microglia (1 X 10⁴ cells/well) was assessed using the linear target electrode, placed between cell and chemoattractant wells. (B) Principles of Boyden chemotaxis chamber (left). Microglial migration toward 2-AG-containing lower wells of the Boyden chamber in the absence or presence of hinokiresinols [10 μM each (middle) or indicated concentration of <i>trans</i>-hinokiresinols (right)] was quantified, and the results are expressed as fold change in numbers of migrated cells (left) or an inhibition percentage compared to basal migration and maximum migration by 2-AG (right), respectively. Values are mean ± SEM of 4–8 independent quantification of migration. ^*<i>P</i> < 0.05: vs. indicated group.</p

Hinokiresinols modulates 2-AG-induced mitochondrial bioenergetics in microglia.

<p>Pure microglia (5 X 10⁵ cells/ml, 150 μl/well) were treated with vehicle or hinokiresinols (HRs; line a), oligomycin (line b), FCCP (line c), and rotenone (line d) at indicated times and the rate of oxygen consumption (OCR, left) and extracellular acidification (ECAR, right) were recorded. (A) Real-time mitochondrial respiration was measured in microglia using the Seahorse analyzer. (B, C) ATP turnover and respiration capacity were calculated based on OCR (A, left panel) and expressed as % of control. The data were normalized in vehicle controls and expressed as mean ± SEM (N = 4–8). ^*<i>P</i> <0.05: vs. indicated group.</p