No abstract

This work aimed to evaluate the activity of a lipid transfer protein isolated from Morinda citrifolia L. seeds, McLTP 1 , on the development of intestinal mucositis following irinotecan administration. McLTP 1 (0.5, 2 and 8 mg/kg, i.v.) was injected into mice 1h before irinotecan administration (75 mg/kg, i.p.; 4 days), and then for additional 6 days. Seven days after the rst dose of irinotecan, diarrhea was assessed and the intestine was removed for histological evaluation, assessment of intestinal over-contractility, measurement of myeloperoxidase (MPO), glutathione (GSH), malondialdehyde (MDA), proin ammatory cytokines and chemokine (IL-1, IL-6, and KC levels -a murine homolog of human IL-8 chemokine), analysis of cyclooxygenase 2 (COX-2), nuclear factor kappa B (NF-κB) and nitric oxide synthase (iNOS) expression. At the two highest doses, McLTP 1 administration decreased mortality and diarrhea. McLTP 1 (8 mg/kg, i.v.) signi cantly prevented irinotecan-induced intestinal damage and led to a reduction in overcontractility of the intestinal muscle (p < 0.05). Moreover, McLTP 1 decreased the MPO, IL-1β, IL-6, and KC levels signi cantly by 74.7%, 42%, 92.9%, and 95.9%, respectively. Also, the expression of COX-2, NF-κB, and iNOS was reduced. Pretreatment with McLTP 1 (8mg/kg; i.v.) signi cantly reduced the MDA level and increased the duodenum homogenates' GSH level. Our study provides a potential new therapeutic for preventing irinotecan-induced mucositis, improved clinical parameters, reduced in ammation, and oxidative damage.

Himatanthus drasticus (Apocynaceae), a laticifer plant, is popularly known as janaguba. This latex has been widely used in traditional folk medicine to treat diseases such cancer and ulcer. This study was performed to investigate the gastroprotective activity of proteins isolated from latex Himatanthus drasticus (HdPL) in gastric ulcer model. Experimental protocols were registered on the Institutional Ethics Committee under number 43/2011. Swiss male mice were treated with PrLP at doses 0.5; 5 and 50 mg/kg (i.v.; n=8). After 30 min they received 0.2 ml of absolute ethanol per oral, after 60 min, the animals were sacrificed, stomachs removed and analyzed the lesion index and dosage of GSH (reduced glutathione) and nitrite. To study the gastroprotective mechanism, its relations to capsaicin‐sensitive fibers, endogenous prostaglandins, nitric oxide, ATP‐sensitive potassium channels and cGMP were analyzed. HdPL gastroprotective effect was observed only at dose of 5mg/kg. HdPL also restored the GSH levels and nitrite in mucosa and only L‐NAME, Glibenclamide and ODQ were able to reverse the protective effect of PrLP. We can conclude that the HdLP has pharmacological activity with gastroprotetor effect in the gastric mucosa. This protection appears to be mediated in part by modulation of NO/cGMP/KATP pathway, which has been related to mucosal defense and maintenance of stomach's blood flow. Support: CNPQ ans CAPES.

The larger number of plants, with therapeutic potential, popularly used in Northeastern Brazil is due to their easy access and the great Brazilian biodiversity. Previously, was demonstrated that the methanol fraction from Sideroxylon obtusifolium (MFSOL) promoted an anti-inflammatory and healing activity in excisional wounds. Thus, this work aimed to investigate the healing effects of MFSOL on human keratinocytes cells (HaCaT) and experimental burn model injuries. HaCaT cells were used to investigate migration and proliferation of cell rates. Female Swiss mice were subjected to seconddegree superficial burn protocol and divided into four treatment groups: Vehicle (cream-base), 1.0% Silver Sulfadiazine (Sulfa), and 0.5% or 1.0% MFSOL cream (CrMFSOL). Samples were collected for quantification of the inflammatory mediators and histological analyses after 3, 7 and 14 days on evaluation. As result, MFSOL (50 μg/ml) stimulated HaCaT cells by increasing proliferation and migration rates.Moreover, CrMFSOL 0.5% attenuated myeloperoxidase (MPO) activity and also stimulated the release of IL-1β and IL-10, after 3 days with treatment. CrMFSOL 0.5% enhanced wound contraction, promoted tissue remodeling improvement and highest collagen production after 7 days, and VEGF release after 14 days. Therefore, MFSOL evidenced the stimulation of human keratinocyte (HaCaT) cells and improvements on wound healing via inflammatory modulation on burn injuries.

Burns are health problems that overwhelm the Unified Health System (SUS) in Brazil. Despite the new therapeutic strategies, the costs of treating burns ate still quite high, and there are no effective alternatives for healing the skin. The use of plants with therapeutic potential is popularly used, due to its low cost, easy access and great Brazilian biodiversity. McLTP1, a lipid transfer protein isolated from Morinda citrifollia (noni) seeds, has shown antinociceptive, anti-inflammatory, antibacterial and antioxidative effects. Therefore, the aim of this study was to investigate the effect of McLTP1 on the healing of superficial burns in mice. The study was approved by CEUA NPDM UFC (protocol: 021706190). The burn was induced by direct contact with a square stainless-steel plate (1.5 cm2). The animals were divided into five experimental groups (n=6-7/grupo) and treated daily with 0.9% NaCl saline solution (Sham), or with topical treatment performed with dermatological creams: Silver sulfadiazine 1% (Sulfa 1%), lanette cream (Vehicle), cream lanette containing 0.25% and 0.5% of McLTP1. The animals were euthanized after 14 days. McLTP1 promoted total wound closure after 2 weeks of treatment, reduced histopathological scores at 3rd day, as well as induced the formation of a thicker epithelium and collagens synthesis on 14th day, modulated inflammation by reducing MPO activity, TNF-a, IL-1B and IL-6 levels and increasing IL-10 after 3 days of burn, modulated VEGF production at three times analyzed in this study, increased TGF-B and immunostaining for FGF after 7 days, reduced immunostaining for TNF-a on the 3rd day and exerted an antioxidant function by reducing MDA and nitrite and increasing GSH at day 3. In short, McLTP1 showed an important healing action in this burn model, showing additional anti-inflammatory and antioxidant effects.

This work aimed to evaluate the activity of a lipid transfer protein isolated from Morinda citrifolia L. seeds, McLTP1, on the development of intestinal mucositis following irinotecan administration. McLTP1 (0.5, 2 and 8 mg/kg, i.v.) was injected into mice 1h before irinotecan administration (75 mg/kg, i.p.; 4 days), and then for additional 6 days. Seven days after the first dose of irinotecan, diarrhea was assessed and the intestine was removed for histological evaluation, assessment of intestinal over-contractility, measurement of myeloperoxidase (MPO), glutathione (GSH), malondialdehyde (MDA), proinflammatory cytokines and chemokine (IL-1, IL-6, and KC levels - a murine homolog of human IL-8 chemokine), analysis of cyclooxygenase 2 (COX-2), nuclear factor kappa B (NF-κB) and nitric oxide synthase (iNOS) expression. At the two highest doses, McLTP1 administration decreased mortality and diarrhea. McLTP1 (8 mg/kg, i.v.) significantly prevented irinotecan-induced intestinal damage and led to a reduction in over-contractility of the intestinal muscle (p < 0.05). Moreover, McLTP1 decreased the MPO, IL-1β, IL-6, and KC levels significantly by 74.7%, 42%, 92.9%, and 95.9%, respectively. Also, the expression of COX-2, NF-κB, and iNOS was reduced. Pretreatment with McLTP1 (8mg/kg; i.v.) significantly reduced the MDA level and increased the duodenum homogenates' GSH level. Our study provides a potential new therapeutic for preventing irinotecan-induced mucositis, improved clinical parameters, reduced inflammation, and oxidative damage.

Hemorrhagic cystitis is a side effect of chemotherapy induced by an antineoplastic agent from the oxazaphosphorine group (ifosfamide and cyclophosphamide), resulting from the formation of the urotoxic metabolite acrolein. Morinda citrifolia Linn., popularly known as noni, is a species of Rubiaceae, where it is used from the root to the fruit for therapeutic purposes. From the seeds, a thermostable protein called McLTP1 (9.4 kDa) was extracted, among its therapeutic effects, it showed anti-inflammatory, gastroprotective, antibacterial and antinociceptive activity. Thus, the objective of this study is to evaluate the protective effect and the possible mechanism of action of a protein isolated from the seed of Morinda citrifolia (McLTP1) in hemorrhagic cystitis induced by ifosfamide in mice. Hemorrhagic cystitis was induced by intraperitoneal (i.p) administration of ifosfamide (IFO) in a single dose of 400mg/kg, according to a standardized protocol, in male balb/c mice. The experimental group treated with the uroprotective drug, mesna (80 mg/kg; i.p), received a pretreatment 30 minutes before, 4 and 8 hours after IFO. Treatment with McLTP1 was divided into two protocols, the first to define the best dose through a dose-response curve, where a pre-treatment was performed three days before cystitis induction, with McLTP1 administered at doses of 10, 20 or 40mg/kg (i.p), and two treatments 2 and 4 hours after IFO administration, evaluating its effect on bladder wet weight, edema and hemorrhage scores, and neutrophilic infiltrate. In the second protocol, only the best dose was used for the analysis of its effect on the hemorrhagic cystitis model. After 12 hours of hemorrhagic cystitis induction, the animals were euthanized by a high anesthetic dose. Subsequently, the bladders were removed, weighed and kept in 10 per cent buffered formalin for histological, immunohistochemical (COX-2 and TNF-a), immunofluorescence (NF-kB and F4-80) analyses, or stored at -80C for of MPO, vascular permeability, hemoblobin, cytokines (TNF-a;, IL-1B;, IL-6, IL-10, IL-4, IL-33), enzymes (iNOS and COX-2) and markers of oxidative stress (MDA, NO, GSH, SOD and CAT). The adopted experimental procedures were approved by the Animal Research Ethics Committee through protocol number 23170920-0. Treatment with McLTP1 reduced bladder wet weight at the three respective doses mentioned above, however, it was observed the reduction of toxicity parameters (macroscopic edema and hemorrhage scores) only at the lowest dose (10 mg/kg), as well as MPO activity at doses of 10 and 20 mg/kg (p<0.05). results, the lowest dose was chosen for subsequent results. McLTP1 (10 mg/kg) was able to promote permeability reduction and vascular and hemoglobin in the bladder through quantification by the evans blue method and cyanmethemoglobin, respectively (p<0.05). In addition, it had a protective effect by attenuating inflammatory scores and preserving the structure of the urothelium. The anti-inflammatory activity was demonstrated through the significant decrease of the cytokines TNF-a; IL-B;, IL-6 and increase of IL-10; reduced expression of COX-2, NF-kB and F4/80, and gene expression of IL-33, IL-4 and iNOS (p<0.05). McLTP1 also showed antioxidant activity, being able to reduce MDA and NO and increase levels of GSH, SOD and CAT (p<0.05). From the presented data, we can infer that McLTP1 is a potential uroprotector in the prevention of ifosfamide-induced hemorrhagic cystitis in mice by reducing inflammatory parameters and antioxidant activity.

Background Interstitial lung disease (ILD) and pulmonary arterial hypertension increase morbidity and mortality of systemic sclerosis (SSc) patients [1]. Cyclophosphamide (CYC) has been demonstrated to be efficacious in forced expiratory volume (FEV1), forced vital capacity (FVC) and diffusing capacity of the lung for carbon monoxide (DLCO) improvement, however few real-life long-term observations have been reported [2.3]. Objectives Our goal is to report CYC effectiveness and safety for ILD treatment in SSc. Methods All patients from a University-based SSc clinic that used intravenous (IV) CYC for ILD treatment between January 2003 and December 2013 were included. Data were extracted from medical records and standardized forms that were completed during every clinical visit. Outcome measures were FEV1, FVC and DLCO variations before and up to 6 months after CYC treatment, and severe adverse events during CYC use. Results Twenty-eight patients were included. A total of 36 monthly IV CYC treatments were observed. Patients were mostly middle-age (mean ± SD) (49.7±14.2 years), white (86%), women (86%), with limited cutaneous SSc (64%) and disease duration of 3.4±3.9 years. Before treatment lung tests values were FEV1 70.4±19.5%, FVC 68.7±19.4%, DLCO 58.3±17.0ml/min/mmHg. Values for IV CYC were dose 924.5±210.1mg, treatment duration 16.1±18.3 months, infusions (number per treatment) 8.4±3.6. Lung tests variations after CYC treatments were ΔFEV1 2.46±9.54% (P=0.15), ΔFVC 2.78±9.62% (P=0.11), ΔDLCO -2,39±8.65ml/min/mmHg (P=0.09). Fourteen out of 36 CYC treatments (39%) improved above the minimal clinically important difference of 6% in FVC, whereas 7 (19%) worsened. Seventeen severe adverse events were reported (0.47 severe adverse event per CYC treatment), mostly infections. The cumulative CYC dose for each severe adverse event was 2,687±1,779mg. Seven hospital admissions (5.0±4.6months after first CYC infusion) and 3 deaths (8.3±8.7months after last CYC infusion) were observed. Conclusions In our study, CYC treatment did not change FEV1, FVC and DLCO. In almost half of CYC treatments a severe adverse event was reported. Prolonging CYC treatment may increase severe adverse events risk at no additional effectiveness. Other long-term studies are necessary for finding the optimum CYC treatment dose and duration in SSc lung disease. References Steen VD, Conte C, Owens GR, Medsger TA Jr. Severe restrictive lung disease in systemic sclerosis. Arthritis Rheum1994; 37:1283-9. Tashkin DP, Elashoff R, Clements PJ, et al. Effects of 1-year treatment with cyclophosphamide on outcomes at 2 years in scleroderma lung disease. Am J Respir Crit Care Med. 2007; 176:1026– 1034. Apras S, Ertenli I, Ozbalkan Z, et al. Effects of oral cyclophosphamide and prednisolone therapyon the endothelial functions and clinical findings in patients with early diffuse systemic sclerosis. Arthritis Rheum. 2003;48:2256-2261. Disclosure of Interest None declared DOI 10.1136/annrheumdis-2014-eular.3044