Cholinergic agonists inhibit proliferation of human fibroblast-like synoviocytes and monocytic cell lines and reduce VEGF and MMPs expression by these cells.

Background and purpose: Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by inflammation and joint destruction. Excessive proliferation of fibroblast-like synoviocytes (FLS) and over-expression of angiogenic factors play a crucial role in pannus formation and joint destruction in RA. Clarification of the role of cholinergic agonists in modulation of inflammation and immune system reactions is progressively ongoing. In this study, the anti-angiogenic effect of two cholinergic agonists, nicotine and ARR17779, on human FLS, and monocytic cell lines (U937) was evaluated.Experimental approach: The cells were cultured in DMEM supplemented with 10% FBS and treated with different doses of nicotine and ARR17779 in the presence of TNF-α, LPS, and IFN-γ. After 48 h, cell number was counted in different groups. After RNA extraction, cDNA was synthesized and the expression of VEGF and MMPs has been evaluated by real-time PCR using specific primers and probes. VEGF was assayed in U937 cell line supernatant using ELISA method.Key results: Both nicotine and ARR17779 inhibited FLS and U937 cell proliferation. Cholinergic agonists reduced the expression of MMPs and VEGF. VEGF level in supernatant of U937 cells treated with cholinergic agonists was also reduced.Conclusion and implications: Our results suggest that cholinergic agonists can modulate pathological conditions related to pannus formation in in-vitro conditions. Based on these results, cholinergic agonists can be considered as novel therapeutic options in RA. Further animal studies are needed before introducing these agents into clinical uses.

Plasma level of neopterin as a marker of disease activity in treated rheumatoid arthritis patients: association with gender, disease activity and anti-CCP antibody.

Immune system activation is known to be involved in the progression of rheumatoid arthritis (RA). The pro-inflammatory cytokine interferon-γ in various cells, including monocytes, induces neopterin production. Plasma level of neopterin has been measured in many autoimmune diseases and can be used as a marker of cellular immunity activation. In this study we measured the plasma level of neopterin in 418 treated RA patients and 398 age and sex matched healthy people by high pressure liquid chromatography (HPLC) method. Disease activity score was calculated in all patients by DAS-CRP method. Plasma level of neopterin was compared between RA and control groups. We also determined the association between neopterin level with gender and disease activity score in RA patients. Significantly higher level of neopterin was observed in RA patients compared to healthy controls. Moreover, there was higher neopterin level in male RA patients versus female patients. Plasma neopterin level was increased in patients with active disease and also was correlated with disease activity parameters. There was a significant correlation of plasma level of neopterin with age in both RA and control group and also age of onset and disease duration in RA patients. Anti-CCP positive patients had higher level of neopterin in comparison to anti-CCP negative patients and there was a significant correlation between neopterin level and anti-CCP titer. Our results indicated that neopterin is a sensitive marker for assaying background inflammation and disease activity score in RA patients and may be used as a marker for evaluation of therapy efficacy.

Protective effect of pomegranate seed oil against acute toxicity of diazinon in rat kidney.

Diazinon is an organophosphate which is extensively used in trade and agriculture. Due to its widespread application, its toxicity is common. Several studies have shown that organophosphates are able to induce oxidative stress by generating free radicals and depletion of endogenous antioxidants. Pomegranate seed oil (PSO) possesses anti-inflammatory and antioxidant effects. In this study, the effect of PSO was evaluated on diazinon-induced nephrotoxicity in rat. Wistar male rats were randomly divided into four groups, 6 each. Group one received saline, 1 mL/kg, group 2 received diazinon 100 mg/kg. Groups 3 and 4 received PSO, 0.32 and 0.64 mg/kg, one hour before diazinon 100 mg/kg respectively. After 24 h, animals were anesthetized. Blood samples were taken out by cardiac puncture for measuring the level of serum urea and creatinine. 24 h urine samples were also collected for measuring glucose and protein concentration. The right kidney was removed and homogenized for measuring malondialdehyde and thiol content Compare to control group, DIZ increased urea and serum creatinine, urinary glucose, and malondialdehyde, but did not modify significantly urinary protein and thiol content. In groups received PSO+ DIZ, serum creatinine, urinary glucose and MDA were significantly decreased. DIZ induced acute nephrotoxicity and oxidative stress. Probably, increasing of serum creatinine and urinary glucose are appropriate markers for diagnosis of kidney damage. In addition increasing of MDA level emphasizes that DIZ plays role in pathogenesis of kidney via oxidative stress mechanism. PSO reduced DIZ toxicity by antioxidant activity.

In vitro and in vivo anti-angiogenesis effect of shallot (Allium ascalonicum): a heat-stable and flavonoid-rich fraction of shallot extract potently inhibits angiogenesis.

This study has been undertaken to elucidate the anti-angiogenic properties of shallot extract in vitro and in vivo and also to define the responsible fraction and its stability. After preparation of the extract of shallot bulbs with 50% ethanol, the extract was successively fractionated into n-hexane, ethyl acetate, n-butanol and aqueous fractions. The ethyl acetate fraction was further fractionated to three subfractions using thin layer chromatography. Anti-angiogenic activity of fractions and subfractions were examined on human umbilical vein endothelial cells (HUVECs) in collagen matrix and chicken chorioallantoic membrane (CAM) models. Among the fractions, ethyl acetate fraction and one of its subfractions potently inhibited angiogenesis in vitro and in vivo. Furthermore, ethyl acetate fraction sustained its inhibitory effect significantly even after treatment in high thermal and low pH conditions. These findings provided a useful basis for further investigations on shallot as a useful herb with therapeutic or preventive activity against angiogenesis related disorders.

Corneal neovascularization: molecular events and therapeutic options.

Corneal neovascularization (NV) is a significant complication of numerous infectious and non-infectious ocular surface disorders. Presence of newly formed blood vessels in the cornea can compromise clarity and therefore vision. Various growth factors and proteinases seem to be involved in the corneal NV. During corneal injury, angiogenic factors are released from corneal epithelial and stromal cells as well as infiltrating immune cells like macrophages. In fact, the balance between angiogenic and anti-angiogenic factors is shifted towards angiogenic molecules in the corneal NV. Numerous investigations support this idea that vascular endothelial growth factor (VEGF) plays a pivotal role in corneal NV by inducing endothelial cells proliferation, migration, and tubulogenesis. There is a growing body of evidence that corneal NV can be reduced by using anti-VEGF agents. This article reviews the most known molecular events in corneal NV and also some of the recent patents relevant to the field. Understanding the role of growth factors, proteinases and inflammatory cytokines in corneal NV can help the investigators to design therapeutic options for controlling this debilitating condition.