Rotula aquatica Lour. mitigates oxidative stress and inflammation in acute pyelonephritic rats.

The current study evaluates the efficacy of methanolic extract of Rotula aquatica Lour. (MERA) against inflammatory changes associated with acute pyelonephritis. The antioxidant enzymes such as SOD, CAT, GPx, GR and oxidative stress markers like GSH content, malondialdehyde (MDA) level, nitrate level, reactive oxygen species (ROS) level and renal toxicity markers were evaluated in this study. The mRNA level expression of Toll-like receptor 4 (TLR-4), nuclear transcription factor kappa B (NF-κB), tumour necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and Tamm Horsfall protein (THP) were studied by RT-PCR analysis. The oral administration of MERA increases the antioxidant enzyme status in pyelonephritis rat. The elevated levels of oxidative stress markers in pyelonephritic rats were ameliorated by the administration of MERA at 100 mg/kg and 200 mg/kg bwt of the rat. The mRNA level expression of major genes were restored to normal level by MERA.

Acute oral toxicity and anti-inflammatory evaluation of methanolic extract of Rotula aquatica roots in Wistar rats.

BACKGROUND: The plant Rotula aquatica Lour. was traditionally well known due to its large number of pharmacological action and medicinal uses. The plant is a necessary component of many Ayurvedic drug preparations since historical times. It is widely used as a crucial ancient drug for kidney and bladder stones. OBJECTIVES: The main objective of the study was to evaluate the acute toxicity and anti inflammatory efficacy of methanolic extract of R.aquatica Lour. in in vivo models. MATERIALS AND METHODS: The qualitative phytochemical analysis and invitro antioxidant activity of the roots of methanolic extract of R.aquatica Lour. (MERA) was evaluated. The acute toxicity effect of MERA was evaluated with two different doses (550, 2000 mg/kg body weight), were administrated orally to Wistar rats. The rats were observed for sign and symptoms of toxicity and mortality for 14 days. The parameters measured including relative organ weight, blood, biochemical and histopathological parameters of hepatic and renal toxicity. The anti-inflammatory effect of MERA was also evaluated in carrageenan and dextran-induced paw edema models. RESULTS: The phytochemical evaluation of MERA shows the presence of secondary metabolites like alkaloids, flavonoids, phenolics and tannins, phytosterols, reducing sugars, proteins and terpenoids. The results of in-vitro antioxidant evaluation of MERA reveal its capability to scavenging free radical at a lower concentration. The MERA did not show any visible signs of toxicity up to the dose of 2000 mg/kg body weight. The results obtained from our carrageenan and dextran-induced paw edema model study also proved the anti-inflammatory effect of MERA in rat model. CONCLUSION: The result shows the potential of MERA as an anti-inflammatory drug to reduce the signs of inflammation devoid of any toxic effect.

Rotula aquatica Lour. inhibits growth and biofilm formation of clinically isolated uropathogenic Escherichia coli

Objective: To evaluate the anti-bacterial and anti-biofilm activity of ethyl acetate fraction of Rotula aquatica Lour. (EFRA) against clinically isolated uropathogenic Escherichia coli. Methods: In vitro antibacterial and anti-biofilm studies were employed. The antimicrobial activity of EFRA was assayed by the well diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the active fraction were determined by Resazurin method. The time-kill kinetic assay, acridine orange-ethidium bromide staining, propidium iodide uptake assay, and scanning electron microscopic (SEM) analysis were done to evaluate the efficacy of EFRA in killing uropathogenic Escherichia coli. The anti-biofilm activity was determined by 3-[4,5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium-bromide (MTT) assay and specific biofilm formation assay. Results: The well diffusion assay of EFRA showed a very clear zone of inhibition against Escherichia coli BRL-17. The MIC and MBC of EFRA were 2.5 mg/mL and 5 mg/mL, respectively. The time-kill kinetic assay, fluorescence microscopic analysis, propidium iodide uptake assay, and SEM analysis displayed the effect of EFRA in killing the bacteria. The MTT assay and specific biofilm formation assay showed that EFRA prevented the formation of biofilms. Conclusions: The results of the present study confirm that EFRA could prevent bacterial growth and inhibit its biofilm formation.

Rotula aquatica Lour. inhibits growth and biofilm formation of clinically isolated uropathogenic Escherichia coli

Objective: To evaluate the anti-bacteria! and anti-biofilm activity of ethyl acetate fraction of Rotula aquatica Lour. (EFRA) against clinically isolated uropathogenic Escherichia coli. Methods: In vitro antibacterial and anti-biofilm studies were employed. The antimicrobial activity of EFRA was assayed by the well diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the active fraction were determined by Resazurin method. The time-kill kinetic assay, acridine orange-ethidium bromide staining, propidium iodide uptake assay, and scanning electron microscopic (SEM) analysis were done to evaluate the efficacy of EFRA in killing uropathogenic Escherichia coli. The anti-biofilm activity was determined by 3-[4,5- dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium-bromide (MTT) assay and specific biofilm formation assay. Results: The well diffusion assay of EFRA showed a very clear zone of inhibition against Escherichia coli BRL-17. The MIC and MBC of EFRA were 2.5 mg/mL and 5 mg/mL, respectively. The time-kill kinetic assay, fluorescence microscopic analysis, propidium iodide uptake assay, and SEM analysis displayed the effect of EFRA in killing the bacteria. The MTT assay and specific biofilm formation assay showed that EFRA prevented the formation of biofilms. Conclusions: The results of the present study confirm that EFRA could prevent bacterial growth and inhibit its biofilm formation.

Protective effect of Rotula aquatica Lour against gentamicin induced oxidative stress and nephrotoxicity in Wistar rats.

Gentamicin is an aminoglycoside antibiotic widely used for the treatment of life-threatening infections caused by Gram-negative bacteria. The use of gentamicin was limited due to its ototoxic and nephrotoxic adverse effects. The current study was designed to evaluate the protective effect of ethyl acetate fraction from Rotula aquatica (EFRA) against gentamicin induced nephrotoxicity. The antioxidant enzymes status, lipid peroxidation, nitrate and ROS level, serum markers like creatinine, Urea, BUN were estimated in the present study. The histopathological analysis of renal tissues was done by H&E and PAS staining. The mRNA level expression of KIM-1, NF-κB, TNF- α, and IL-6 were measured by semi-quantitative reverse transcription-polymerase chain reaction. The changes in antioxidant parameters were restored by the treatment of EFRA at different dose (50 mg/kg bwt, 100 mg/kg bwt). The serum parameters, ROS, MDA and nitrate level were decreased by administration of EFRA. The EFRA ameliorates histological changes associated with gentamicin induced nephrotoxicity. The mRNA level expression of KIM-1, NF-κB, TNF- α, and IL-6 were downregulated in EFRA treated groups. The results from present study reveals the role of EFRA as good anti-inflammatory and nephro protective drug.

Rotula aquatica Lour attenuates secretion of proinflammatory mediators and cytokines in lipopolysaccharide-induced inflammatory responses in murine RAW 264.7 macrophages.

Rotula aquatica belongs to the family Boraginaceae, and is reported to contain baunerol, steroids and alkaloids. In Ayurveda, R. aquatica has been used for the treatment of various diseases such as diabetes, treatment of piles, venereal disease, and cancer. The current study aims to investigate the anti-inflammatory effect of methanolic extract of R. aquatica (MERA) in RAW 264.7 cells. The cytotoxicity of MERA was analyzed by MTT assay. The total cyclooxygenase (COX) activity, 5-lipoxygenase (5-LOX) activity, myeloperoxidase activity, inducible nitric oxide synthase activity, nitrate level and reactive oxygen species production were studied in LPS-stimulated RAW 264.7 cells. The gene level expression of cyclooxygenase-2 (COX-2), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6) were also evaluated in this study. The MERA did not show any cytotoxicity at different concentrations (6.25-100 µg/ml). MERA (100 µg/ml) inhibited total COX and 5-LOX activity at 50.53 and 62.03%, respectively, besides significantly (p < 0.05) diminished nitrate and ROS generation, when compared with LPS control. Moreover, MERA down-regulated the mRNA expressions of inflammatory marker genes like TNF-α, IL-6, and COX-2 against LPS stimulation. Our results demonstrate that MERA is able to attenuate inflammatory response, possibly via ROS and NO suppression, inhibiting the production of arachidonic acid metabolites and modulation of proinflammatory mediators and cytokines release.

In silico analysis of aqueous root extract of Rotula aquatica Lour for docking analysis of the compound 3-O-acetyl-11-keto-ß-boswellic acid contents.

Molecular docking is a bioinformatics tool used to study and analyse ligand receptor interactions. This helps in identifying the receptors (molecular targets) for different ligands. Using these technologies, compound isolation and drug discovery from herbals is achieved. Herbs are widely used in treatment of various ailments from time immemorial. Phytochemists and drug developers are now interestingly working in developing new molecules that can act effectively than conventional drugs. As they are developing it mostly from herbs they are found to be effective and safer drugs and quantity to be used become minimum. Rotula aquatica Lour is a plant distributed widely in India and used for urinary disorders. The plant root was extracted and studied for its active compounds that possess antiurolithiatic activity. After performing various preliminary phytochemical studies and applying chromatographic methods, molecular docking was carried out with isolated bioactive compound and Tamm-Horsfall protein (THP). By docking analysis the bioactive compound 3-O-acetyl-11-keto-ß-boswellic acid interacted with THP and it may inhibit calcium oxalate crystallization.