Nitrodi thermal water downregulates protein S‑nitrosylation in RKO cells

Abstract

Balneotherapy and spa therapy have been used in the treatment of ailments since time immemorial. Moreover, there is evidence to suggest that the beneficial effects of thermal water continue for months following the completion of treatment. The mechanisms through which thermal water exerts its healing effects remain unknown. Both balneological and hydroponic therapy at 'the oldest spa in the world', namely, the Nitrodi spring on the Island of Ischia (Southern Italy) are effective in a number of diseases and conditions. The aim of the present study was to investigate the molecular basis underlying the therapeutic effects of Nitrodi spring water in low-grade inflammation and stress-related conditions. For this purpose, an in vitro model was devised in which RKO colorectal adenocarcinoma cells were treated with phosphate-buffered saline or phosphate-buffered saline prepared with Nitrodi water for 4 h daily, 5 days a week for 6 weeks. The RKO cells were then subjected to the following assays: 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay, Transwell migration assay, western blot analysis, the fluorimetric detection of protein S-nitrosothiols and S-nitrosylation western blot analysis. The results revealed that Nitrodi spring water promoted cell migration and cell viability, and downregulated protein S-nitrosylation, probably also the nitrosylated active form of the cyclooxygenase (cOX)-2 protein. These results concur with all the previously reported therapeutic properties of Nitrodi spring water, and thus reinforce the concept that this natural resource is an important complementary therapy to traditional medicine.

Water from Nitrodi’s Spring Induces Dermal Fibroblast and Keratinocyte Activation, Thus Promoting Wound Repair in the Skin: An In Vitro Study

“…The main components found were chloride (Cl − ) with a mean value of 93 mg/L, sulfates (SO 4 2− ) with a mean value of 204 mg/L, sodium (Na + ) with a mean value of 174 mg/L, and calcium (Ca 2+ ) with a mean value of 137 mg/L. These results obtained were comparable with those reported by Aversano, [ 27 ], which indicated an amount of 0.2200 g/L (220 mg/L), 0.1676 g/L (167.6 mg/L), and 0.1182 g/L (118.2 mg/L), for SO 4 2− , Na + , and Ca 2+ , respectively. However, the HCO 3 − concentration obtained in this study (561 mg/L) was higher than that reported by Aversano (475.8 mg/L).…”

The Benefits of Water from Nitrodi’s Spring: The In Vitro Studies Leading the Potential Clinical Applications

“…Firstly, the phenomenon of S-nitrosylation, understood as the addition of nitric oxide (NO) to the thiol side chain of cysteine residues within proteins, has been the subject of a lot of academic interest. The aberrant S-nitrosylation of proteins is associated with many diseases [ 61 ]. NO is also a key messenger in the pathogenesis of inflammation and exerts this effect by activating innate and adaptive immunity.…”

“…The results obtained from these experiments showed that MMP-2 protein expression and protein S-nitrosylation were markedly downregulated in RKO cells treated with Nitrodi’s water. Overall, the results revealed that the molecular mechanisms through which Nitrodi’s spring water exerts its anti-inflammatory effects might involve the downregulation of protein S-nitrosylation [ 61 ].…”

Modulation of wound healing regulators by thermal spring water from La Bourboule

“…These results represent early evidence that sulphur-based compounds may be useful in treating chronic in ammatory ailments, despite the mechanism remaining unknown. More recently, Aversano et al reported that a thermal mineral water showed an in-vitro anti-in ammatory activity by down-modulating cyclooxygenase (COX)-2 and matrix metallopeptidase (MMP) 2 proteins (Aversano et al 2020). In ammation is an important step of wound healing and begins after injury.…”