Anti-inflammatory effects of lavender and eucalyptus essential oils on the in vitro cell culture model of bladder pain syndrome using T24 cells

Abstract

Background Interstitial cystitis (IC) has a chronic chemical irritation and inflammation of non-bacterial origin in the bladder wall leading to various severe symptoms. There is evidence that chronic inflammation is significantly associated with abnormal urothelial barrier function, epithelial dysfunction. This is the underlying cause of urothelial apoptosis and sterile inflammation. Method The anti-inflammatory effects of lavender and eucalyptus essential oils (EOs) and their main components (linalool and eucalyptol) were investigated in the T24 human bladder epithelial cell line on TNFα stimulated inflammation, at 3 types of treatment schedule. The mRNA of pro-inflammatory cytokines (IL-1β, IL-6, IL-8) were measured by Real Time PCR. Human IL-8 ELISA measurement was performed as well at 3 types of treatment schedule. The effects of lavender and eucalyptus EOs and their main components were compared to the response to NFκB inhibitor ACHP (2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(4-piperidinyl)-3-pyridinecarbonitrile). Result There is no significant difference statistically, but measurements show that lavender EOs are more effective than eucalyptus EO. Long time treatment (24 h) of both lavender EO and linalool showed higher effect in decreasing pro-inflammatory cytokine mRNA expression than ACHP inhibitor following TNFα pre-treatment. Moreover, both lavender EOs were found to be significantly more effective in decreasing IL-8 secretion of T24 cells after TNFα pre-treatment compared to the ACHP NFκB-inhibitor. Conclusion The lavender EOs may be suitable for use as an adjunct to intravesical therapy of IC. Their anti-inflammatory effect could well complement glycosaminoglycan-regenerative therapy in the urinary bladder after appropriate pharmaceutical formulation.

Comprehensive transcriptome profiling of urothelial cells following TNFα stimulation in an in vitro interstitial cystitis/bladder pain syndrome model

“…As normal human bladder urothelial cells are difficult to obtain for ethical reasons, while cell lines are widely accessible, our in vitro model consisted of the human non-invasive bladder cancer urothelial cell line RT4. In contrast to the more commonly used muscle-invasive bladder cancer urothelial cell line T24 in previously published in vitro IC/BPS models ( 18 , 20 , 21 , 23 ), RT4 cells isolated from grade I transitional cell carcinoma better reflect normal urothelial cells, especially with respect to differentiation markers ( 24 , 25 ), as well as growth and migration characteristics ( 26 ). However, to increase the credibility of our results, we also confirmed our main findings from RT4 cells in normal porcine bladder urothelial (NPU) cells, which are more accessible and share most structural, molecular, and physiological features with normal human bladder urothelial cells ( 27 – 29 ).…”

“…Furthermore, TNFα signaling pathway has been determined as significantly enriched in the bladders of IC/BPS patients, specifically in those with Hunner’s lesions ( 17 ). It is therefore not surprising that published in vitro models of bladder urothelial cells frequently use TNFα to mimic an inflammatory milieu found in IC/BPS ( 18 – 23 ). However, the exact effects of TNFα on global gene expression and alterations of cellular processes in urothelial cells are still not yet entirely known.…”

“…Hence, we here determined the overall effect of TNFα on the transcriptomic profile of urothelial cells using RNA sequencing (RNA seq) and validated our result using qPCR, ELISA, Western blots, and immunofluorescence. The concentration of TNFα (20 ng/ml) that was used to stimulate urothelial cells in the current study was selected based on previously published in vitro studies ( 18 – 23 ), viability assays and the ability to elicit a maximum measurable inflammatory response with the methods used that reached statistical significance. The ultimate goal of the in vitro model was to recapitulate the most important pathological processes and pathways, found enriched in the bladders of IC/BPS patients, and identify new potential therapeutic targets.…”

Essential Oils and Neuropathic Pain

“…Eucalyptus essential oil has antiviral activity against herpes simplex virus {HSV)-1, HSV-2, influenza A, coxsackie, and rotavirus [ 19 ]. Lavender essential oil has anti-inflammatory properties and work at the cellular level by decreasing pro-inflammatory cytokine mRNA expression and IL-8 secretion [ 20 ]. In studies by Takikawa et al, nutmeg essential oil had antibacterial effects on pathogenic strains of Escherichia coli, and inhibited Yersinia entero-colitica and Listeria monocytogenes growth in broth culture [ 21 ].…”

Extraction of Camphor Tree Essential Oil by Steam Distillation and Supercritical CO2 Extraction

“…Eucalyptus has been used in the preparation of oral condiments as food additives and drug research preparations. Because of its anti-inflammatory effect, it can also improve the lung function of asthmatic rats by intramedullary injection [ 10 ]. Camphor contributes to analgesia and reduces inflammation [ 11 ], has a certain protective effect in mosquito repellents, and has a strong inhibitory effect on mold growth [ 12 ].…”