Biology subjects

Rabelo, L. M. A.

Publications and source records attributed to Rabelo, L. M. A..

3 recordsLinked to original sources

bioRxiv2024

Therapeutic effects of a purified chitin-binding protein from Moringa oleifera seeds on irinotecan-induced intestinal mucositis in mice

Intestinal mucositis is a common side effect of irinotecan-based anticancer regimens, affecting approximately 85% of colorectal cancer (CRC) patients undergoing chemotherapy. Current treatment options are mainly palliative. Moringa oleifera Lamarck, native to Northeast India, is known for its nutritional and therapeutic properties. Our research group has demonstrated that MoCBP4 (11.78 kDa), a thermostable chitin-binding protein isolated from Moringa oleifera seeds, possesses potent antinociceptive, antifungal, wound-healing, and anti-inflammatory activity, both orally and intraperitoneally. Therefore, this study aimed to evaluate the protective effect of Mo-CBP4 in a model of irinotecan (CPT-11)-induced intestinal mucositis. This study was approved by the Animal Research Ethics Committee of UFC - CEPA (9892300120) and (7796300120). Male Swiss mice (25-30 g) were divided into 3 groups: Group 1 received saline solution (0.9%, i.p.) once a day for seven days; Group 2 received irinotecan (75 mg/kg, i.p.) once a day for four days; Group 3 was treated for 7 days with Mo-CBP4 at a dose of 10 mg/kg e.v., respectively, 30 minutes before CPT-11, which was administered for 4 days. During the seven days, weight loss, diarrhea scores, and survival were evaluated. On the seventh day, blood was collected for leukocyte count, followed by euthanasia for duodenum collection and evaluation of the following parameters: small intestine length, intestinal contractility, histopathological and morphometric alterations, MPO, GSH, MDA, NO, cytokines (IL-1{beta}, IL-6, KC, TGF-{beta}, and IL-10), and PCR (IL-33, IL-17, Claudin-2, Occludin, and ZO-1). We evaluated the cytotoxicity of MoCBP4 in normal and tumor cells for 24h and 72h using the Sulforhodamine B (SRB) assay and its interference with the effect of SN38 [2.5M] (active metabolite of CPT-11) for 48h in murine colon cancer (MC38). We injected MC38 cells into male C57BL/6 mice (n=10; 25{+/-}2g) to induce a tumor amenable to treatment with CPT-11. We analyzed daily weight and palpable tumor growth with a digital caliper, and after sacrifice, we measured final tumor growth and MPO. Irinotecan (CPT-11) induced intestinal mucositis in mice, characterized by weight loss, diarrhea, increased mortality, leukopenia, decreased intestinal length, increased intestinal contractility, histopathological alterations (villous blunting, loss of crypt architecture, vacuolation, inflammatory cell infiltrate), and increased levels of MPO, IL-1{beta}, IL-6, KC, TGF-{beta}, IL-33, and Claudin 2. Increased MDA and decreased GSH levels were also observed. Treatment with Mo-CBP4 (10 mg/kg) significantly attenuated CPT-11-induced intestinal mucositis, improving diarrhea, increasing survival, reducing intestinal damage, and attenuating histopathological alterations. MoCBP4 was also able to decrease levels of MPO (35%), NO (48%), IL-1{beta} (52%), IL-6 (98%), KC (88%), TGF-{beta} (62%), IL-33, decrease MDA levels, and increase GSH. MoCBP4 did not exhibit cytotoxic activity in MC38, L929, SK-MEL, and B16F10 cell lines, and did not interfere with the cytotoxic effect of SN38. It also did not interfere with the antitumor effect of CPT-11 in a tumor transplant model. Therefore, Mo-CBP4 demonstrates important antidiarrheal, anti-inflammatory, and antioxidant activities that make it a promising therapeutic option for preventing and attenuating the severity of intestinal mucositis during CPT-11 chemotherapy treatment without interfering with the antitumor effect of irinotecan.

pharmacology and toxicology↗

bioRxiv2023

McLTP1, a lipid transfer protein isolated from noni seeds induces effective healing of superficial burns

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: 02170619-0). 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-, IL-1{beta} 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-{beta} and immunostaining for FGF after 7 days, reduced immunostaining for TNF- 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.

immunology↗

bioRxiv2023

Uroprotective effect of a protein isolated from seed of Morinda citrifolia (McLTP1) on hemorrhagic cystitis induced by ifosfamide in mice

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% buffered formalin for histological, immunohistochemical (COX-2 and TNF-), immunofluorescence (NF-kB and F4-80) analyses, or stored at -80{degrees}C for of MPO, vascular permeability, hemoblobin, cytokines (TNF-, IL-1{beta}, 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-, IL-1{beta}, 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.

pharmacology and toxicology↗