Inhibition of Vascular Smooth Muscle Cell Proliferation by Gentiana lutea Root Extracts

Gentiana lutea belonging to the Gentianaceae family of flowering plants are routinely used in traditional Serbian medicine for their beneficial gastro-intestinal and anti-inflammatory properties. The aim of the study was to determine whether aqueous root extracts of Gentiana lutea consisting of gentiopicroside, gentisin, bellidifolin-8-O-glucoside, demethylbellidifolin-8-O-glucoside, isovitexin, swertiamarin and amarogentin prevents proliferation of aortic smooth muscle cells in response to PDGF-BB. Cell proliferation and cell cycle analysis were performed based on alamar blue assay and propidium iodide labeling respectively. In primary cultures of rat aortic smooth muscle cells (RASMCs), PDGF-BB (20 ng/ml) induced a two-fold increase in cell proliferation which was significantly blocked by the root extract (1 mg/ml). The root extract also prevented the S-phase entry of synchronized cells in response to PDGF. Furthermore, PDGF-BB induced ERK1/2 activation and consequent increase in cellular nitric oxide (NO) levels were also blocked by the extract. These effects of extract were due to blockade of PDGF-BB induced expression of iNOS, cyclin D1 and proliferating cell nuclear antigen (PCNA). Docking analysis of the extract components on MEK1, the upstream ERK1/2 activating kinase using AutoDock4, indicated a likely binding of isovitexin to the inhibitor binding site of MEK1. Experiments performed with purified isovitexin demonstrated that it successfully blocks PDGF-induced ERK1/2 activation and proliferation of RASMCs in cell culture. Thus, Gentiana lutea can provide novel candidates for prevention and treatment of atherosclerosis

­­­A Comparative Evaluation of Antimicrobial Effect of Snuhi-Apamarg-Ksharsutra and Udumbara-Ksheer Sutra in the Management of Bhagandara (Fistula-In-Ano)

Bhagandara one of the eight grave disorders, pertaining the perianal region is arduous to treat, has been described in detail by Acharaya Sushruta. The Ayurvedic description of Bhagandara can be identified as associated with an anal fistula. An anal fistula is defined as an internal opening in the altered skin or mucosa of the anal canal or rectum and an exterior opening in the perianal skin region. Among the various modalities available for the treatment of bhagandara, ksharasutra is described as gold standards for treatment and cure. The ksharasutra possess antimicrobial properties and can be established as first line of treatment for bhagandar(fistula-in-ano). The modern approach for fistula-in-ano is surgical procedure. Antibiotics are commonly used along with surgery for the treatment of fistula-in-ano.  The aim of this study was to comparatively evaluate the antimicrobial properties amongst Snuhi-apamarga kshar sutra and Udumbara ksheer sutra. A total no. of 60 participants were enrolled as per the criteria for the study and pus samples were collected as per the standard operating procedure. Samples of Snuhi-apamarga kshar sutra and Udumbar ksheer sutra were already given in laboratory for testing the sensitivity. Reports were collected and analyzed on different parameters and conclusion was drawn accordingly. It was concluded that gram positive monomicrobial infections were more common especially involving staphylococci. Also, in the culture and sensitivity test Snuhi- apamarga ksharsutra was found to be more effective in comparison to Udumbara ksheer sutra as the microbes were highly sensitive to the Snuhi Apamarga kshar sutra. This can be contributed to the multiple contents in Snuhi Apamarga kshar sutra

<span style="font-size:15.0pt;font-family: "Times New Roman";mso-fareast-font-family:"Times New Roman";mso-bidi-font-family: "Times New Roman";mso-ansi-language:EN-US;mso-fareast-language:EN-US; mso-bidi-language:AR-SA;mso-bidi-font-style:italic" lang="EN-US">Comparative study<span style="font-size:15.0pt;font-family:"Times New Roman";mso-fareast-font-family: "Times New Roman";mso-bidi-font-family:"Times New Roman";mso-ansi-language: EN-US;mso-fareast-language:EN-US;mso-bidi-language:AR-SA" lang="EN-US"> on chemical profiling and antimicrobial properties of essential oils from different parts of <i>Eucalyptus lanceolatus</i></span></span>

425-432<span style="font-size:9.0pt;mso-bidi-font-size: 10.0pt;font-family:" times="" new="" roman";mso-fareast-font-family:"times="" roman";="" mso-bidi-font-family:"times="" roman";mso-ansi-language:en-us;mso-fareast-language:="" en-us;mso-bidi-language:ar-sa"="" lang="EN-US">In this study, essential oils from stem, leaf and bud of Eucalyptus lanceolatus were analysed and chemical profiling was done by Gas Chromatography-Mass Spectrometry (GC-MS). The major components of the oils found to have eucalyptol, alpha pinene, beta pinene, alpha eudesmol and para cymene. Eucalyptol is present in large quantity in bud oil (24.96%) followed by alpha and beta pinene (21.71% and 14.20%, respectively). In stem oil, alpha pinene (32.25%) and beta pinene (19.20%) are present in good quantity. In leaf oil, along with alpha and beta pinene, alpha eudesmol is present in good amount. These essential oils also exhibited potent antifungal and antibacterial activities. Further, antimicrobial and antioxidant constituents of the oils were spotted by bio-autography technique. In addition to this, in vitro DPPH radical scavenging activity and metal ion chelation potential of these oils were also investigated and found out that the bud oil has remarkable scavenging potential with IC50 value of 175µl/ml, followed by leaf and stem oil.</span

Change in bone mineral density in premenopausal women with rheumatoid arthritis managed with or without prednisolone

Background:Osteoporosis (OP) is being increasingly recognized in inflammatory rheumatic diseases like rheumatoid arthritis (RA),&nbsp;characterized mainly by low bone mass, reduced bone strength, and an increased risk of fractures affecting bone metabolism influencing bone mineral density (BMD) and fracture risk.Results :Women in the RA with PRED group did not show lower BMD values than those in the RA without PRED group at baseline, in both lumbar spine L1-L4 (P=0.691), in femoral neck (P=0.332), in radius total (P=0.564) and radius UD (P=0.941). Women in the RA without PRED group had lower T score (Radius UD) (P=0.015) value than those in the RA with PRED group. However, during 12 months follow ups there was no statistically significant difference between the two groups in the change in BMD or projection area in the lumbar spine, femoral neck and radius UD.Conclusion: Premenopausal RA women with or without prednisolone treatment lost their bone mass statistically similar. Study assumes role of RA on axial bone mass development will be less important with better treatment of RA than our patients received

IC₅₀ values for the <i>G. lutea</i> extract (1 mg/ml) on smooth muscle cells.

<p>A) Primary cultures of rat aortic smooth muscle cells (RASMCs), B) Rat aortic smooth muscle cell line A7r5 and C) Human aortic smooth muscle cell line ATCC-CRL-1999.</p

Role of ERK1/2-NO axis on PDGF-induced proliferation of RASMCs and effect of <i>G lutea</i> on PDGF induced NO index.

<p>A) Effect of ERK inhibitor 328000 (1 µmole/L) on PDGF induced cell proliferation, B) Effect of NOS inhibitor L-NAME (10 µmole/L) on PDGF induced cell proliferation, C) Effect of <i>G lutea</i> extract on generation of cellular NO in response to PDGF. *P<0.05 versus control and †<0.05, ††<0.01 and †††<0.001 versus corresponding PDGF treatment.</p

Cell cycle analysis of RASMCs.

<p>A) Representative flow cytogram depicting cells in various stages of cell cycle upon treatment with PDGF-BB (20 ng/ml) in presence and absence of <i>G. lutea</i> extract, and B) Bar graph summarizing data for four independent experiments. ***P<0.001 versus control, ††<0.01 and †††<0.001 versus PDGF treatment.</p

Effect of PDGF-BB (20 ng/ml) on ERK1/2 and NO signaling.

<p>A) Time course of ERK1/2 activation in response to PDGF, B&C) Time course of generation of intracellular nitric oxide in response to PDGF, D) Activation of eNOS through phosphorylation of Ser¹¹⁷⁷ residue in response to PDGF and E) PDGF-induced expression of iNOS. Bar graphs summarize data for a minimum of four independent experiments. *P<0.05, **P<0.01 and ***P<0.001 versus control.</p

Predicted binding energies for <i>G lutea</i> constituents and known MEK1. inhibitors.

<p>Predicted binding energies for <i>G lutea</i> constituents and known MEK1. inhibitors.</p

Effect of isovitexin on PDGF- induced RASMC proliferation.

<p>A) Docking analysis revealing binding pocket of isovitexin on MEK1, B) Effect of isovitexin (0.5–10 µmol/L) on PDGF induced ERK1/2 activation and cell cycle progress and C) Effect of isovitexin on PDGF induced cell proliferation measured through alamar blue assay. Bar graphs summarize data for a minimum of three independent experiments. **P<0.01 versus control and ††<0.01 and †††<0.001 versus PDGF treatment.</p