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This study was designed to characterize muscarinic acetylcholine receptors (mAChRs) in primary cultured Sertoli cells from 30-d-old rats. RT-PCR was performed, and five PCR products corresponding to m1-m5 mAChR mRNA subtypes were detected in these cells. Ribonuclease protection assay further confirmed the presence of protected products for m1, m2, m3, and m4 mAChR transcripts. Radioligand binding studies and the analysis of changes in intracellular signaling pathways after cell exposure to carbachol were performed to study mAChRs at the protein level. Scatchard analysis revealed one single class of [(3)H]quinuclidinyl benzilate binding sites. Carbachol produced a reduction on forskolin-induced intracellular cAMP accumulation in Sertoli cells. This effect was reversed by atropine, methoctramine, and tropicamide but not by p-fluoro-hexahydro-sila-difenidol or pirenzepine. Carbachol also induced an increase on total [(3)H]-inositol phosphates content, an effect antagonized by atropine, p-fluoro-hexahydro-sila-difenidol, or pirenzepine but not by methoctramine. Thus, mAChR activation in Sertoli cell is linked to both adenylyl cyclase inhibition and to phosphoinositide hydrolysis. Furthermore, gel shift assays indicated that carbachol also induced a time-dependent stimulation of the activator protein-1 DNA-binding activity, suggesting that activation of mAChRs may play a role in the modulation of gene expression in Sertoli cells. Taken together, these results indicate that mAChRs are present at mRNA and protein level in rat Sertoli cells.

Abstract:The gastroprotective action of the aqueous extract (AE) and the hydroalcoholic extract obtained from the leaves of Struthanthus marginatus (Desr.) Blume, Loranthaceae, were performed with in vivo models in rodents using: ethanol, indomethacin or stress-induced ulcers, determination of gastric secretion and the mucus production. The scavenger activity of AE in vitro was tested by the DPPH method. The treatment with the extracts (125-1000 mg/kg) significantly inhibited ulcerative lesions in comparison with the negative control groups in all the models evaluated and demonstrated greater effectiveness of the aqueous extract. Regarding the model of gastric secretion, a reduction in volume of gastric juice and total acidity was observed, as well as an increase in the gastric pH. The treatment of rats raised the gastric mucus production. Significant DPPH scavenging activity was evident in the AE. No sign of toxicity was observed. These results show that S. marginatus possesses gastroprotective activity. There are indications that the mechanisms involved in anti-ulcer activity are related to a decrease in acid secretion and an increase in gastric mucus content. Also, there is evidence for the involvement of antioxidant activity in the gastroprotective mechanism.

BackgroundBone defects caused by trauma, infection or tumor resection are common in orthopedic clinic and depending on the extent of the fracture; the vast majority require treatment with bone substitutes. Among the bone grafts employees, the autograft is defined as the gold standard, but with some limitations, principally related to morbidity at the site of its removal, with the need to search for other biomaterials as adjuvant in bone regeneration. Therefore, the objective of the present study was to evaluate the use of Chenopodium ambrosioides as a bone graft substitute for the osseointegration of fractures in rabbits, compared to other bone grafts already employed in the surgical routine as Ricinus communis (castor oil) polyurethane and autogenous bone marrow.MethodsForty-eight rabbits were divided into four groups. After anesthesia, a radius fracture was created and the animals were treated with the following grafts: C. ambrosioides graft, autogenous bone marrow, castor oil and saline (control). After 30, 60 and 90 days, the animals were submitted to radiographical and histological analyses and bone alkaline phosphatase, osteocalcin, biomechanical tension, and collagen were measured. We also realized a phytochemical screening and in vitro antioxidant activity including 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical.ResultsThe data showed that the growth of the bone callus was more expressive and biomechanical assessment showed better tensile strength in C. ambrosioides graft. The experimental results also revealed that there was significantly greater activity of the bone alkaline phosphatase and osteocalcin during early fracture healing, similarly to the group receiving autogenous bone marrow. Histologically, immature bone was observed in C. ambrosioides graft at 30 days, whereas the formation of cartilaginous tissue predominated in the other groups. A higher amount of type I collagen was observed in C. ambrosioides graft throughout treatment. It was detected strong presence of flavonoids and appreciable antioxidant activity.ConclusionsThe results indicate that C. ambrosioides graft and autogenous bone marrow have similar ability to enhance bone regeneration, higher than the castor oil graft, suggesting that the medicinal plant can provide therapeutic benefits for bone tissue.

Chenopodium ambrosioides L. (Amaranthaceae), popularly known as "mastruz" or "erva-de-santa-maria", is a perennial plant found in Brazil and used in folk medicine for the treatment of contusions and fractures. The objective of this study was to evaluate the topical effect of a cataplasm prepared from fresh C. ambrosioides leaves on the treatment of fractures experimentally induced in rabbits. Thirty rabbits were divided into three groups (n = 10). After anesthesia, a radius fracture was created and the animals received topical applications of C. ambrosioides cataplasm (MZG), diclofenac sodium (DG) and isotonic solution 0.9% NaCl (Control). At 30 days, we evaluated the animal bone regeneration through both qualitative macroscopic analysis of the fracture focus as by determination of serum alkaline phosphatase (ALP). The evolution of bone repair has been verified at 30 and 45 days through the histological analysis. No significant difference in ALP levels was observed between groups. In addition, no allergic reactions or impairment of tissue adjacent to the fracture focus were seen in any of the groups. In contrast, although all groups exhibited similar tissue architecture, the histological analysis revealed greater formation of mature bone tissue in MZG at 30 days, when compared to DG and control group. In addition, higher collagen fiber density was observed in MZG at 45 days. These results indicate Chenopodium ambrosioides as a promising therapeutic agent for bone regeneration. The plant may be useful as a raw material for the production of biomaterials for fracture healing, contributing to its validation in ethnomedicine.

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Background. Jacaranda decurrens Cham., known as carobinha, is prevalent in the Cerrado biome and presents popular use in treatment of dermatological diseases. The present study aimed to investigate the healing action of topical formulation of Jacaranda decurrens Cham. (FtEHJ) in mice cutaneous lesions. Methods. Phytochemical analysis of J. decurrens hydroalcoholic extract was carried out by using HPLC-PDA-ESI-MS and FIA-ESI-IT-MSn. Swiss mice were treated topically with formulation base (FtB) or Fibrinase® or ointment FtEHJ (15 mg/g; 50 mg/Kg). At the end of treatment periods, the inflammatory cytokines (TNF-α, IL-1β, and IL-6) in the lesions were measured by using ELISA and gene expression of TGF-β, Collagen I, and Collagen III was demonstrated by RTqPCR method and histological evaluation. Results. Ten compounds were identified in the extract, distributed among the classes of flavonoids and triterpenes. Treatment with FtEHJ increased the wound contraction in 24 hours, such as reduction of TNF-α, IL-1β, and IL-6 (pg/mL) cytokines in the lesion. The TGF-β and collagen gene expression was increased and the wound closure accelerated to nine days, with discrete inflammation, collagenization, and accented reepithelialization. Conclusions. The results obtained suggest chemical compounds present in the FtEHJ accelerates wound healing by being a gene expression modulator, and protein content of different molecules are involved in tissue repair.