Fig 2: Effects of different intensity exercise training on related fibrotic molecules. (A,B) Immunohistochemical staining result and responding quantification of protein products of transient receptor potential vanilloid 4 (TRPV4) and transforming growth factor-beta (TGFβ-1). (C) Bars represent the corresponding protein quantification of TRPV4 and TGFβ-1. Values are mean ± S. D (n = 3 in each group). * p < 0.05 and ** p < 0.01 versus the WKY group. △ p < 0.05 and △△ p < 0.01 versus the SHR group. # p < 0.05 and ## p < 0.01 versus the SHR-H group.

Fig 3: Effects of different intensity exercise training on related fibrotic molecules. (A) Western blotting result and responding quantification of protein products of transient receptor potential vanilloid 4 (TRPV4), transforming growth factor-beta (TGFß-1), anti-Smad2 (phosphoT8) + Smad3 (phosphoT8) (p-Smad2/3), and connective tissue growth factor (CTGF) extracted from the renal tissue. (B) Bars represent the corresponding protein quantification of TRPV4, TGFß-1, p-Smad2/3, and CTGF on the basis of GAPDH. Values are mean ± S.D. (n = 5 in each group). * p < 0.05 and ** p < 0.01 versus the WKY group. ? p < 0.05 and ?? p < 0.01 versus the SHR group. # p < 0.05 and ## p < 0.01 versus the SHR-H group.

Fig 4: Effects of lactic acid on related fibrotic molecules in vitro. (A)Western blotting result and responding quantification of protein products of transient receptor potential vanilloid 4 (TRPV4), transforming growth factor-beta (TGFβ-1), anti-Smad2 (phosphoT8) + Smad3 (phosphoT8) (p-Smad2/3), and connective tissue growth factor (CTGF) extracted from the rat kidney interstitial fibroblast (NRK-49F) cells. (B) Bars represent the corresponding protein quantification of TRPV4, TGFβ-1, p-Smad2/3, and CTGF on the basis of GAPDH. Values are mean ± S.D. (n = 5 in each group). $ p < 0.05 and $$ p < 0.01 versus the lactate group. (C) Immunofluorescence staining of responding protein products of transient receptor potential vanilloid 4 (TRPV4) and transforming growth factor-beta (TGFβ-1).

Fig 5: Novel formulation treatment attenuated inflammatory markers. a Sensitization of skin promoted upregulation of PAR-2, while novel formulation attenuated PAR-2 production. Ears treated with vehicle alone did not develop ACD, while DNFB sensitization (D) triggered upregulation of the itch marker PAR-2. Test substances attenuated the PAR-2 response by 22% and 69%. Typical Western blots are shown. GAPDH, house-keeping protein. D, ACD; MAM + CGDS 2% (formulation A); MAM + CGDS 5% (formulation B). PAR-2 means + SEM, n = 7. *p = 0.002, #p = 0.016. b Skin sensitization promoted upregulation of TRPV4, while novel formulation prevented TRPV4 production. Similarly, ears treated with vehicle alone did not develop ACD; however, DNFB sensitization (D) triggered upregulation of the itch marker TRPV4. Test substance formulation B reduced TRPV4 formation to naïve control levels. Two typical Western blots are shown. GAPDH, house-keeping protein. D, ACD; MAM + CGDS 2% (formulation A); MAM + CGDS 5% (formulation B). TRPV4 means + SEM, n = 7. *p = 0.007, #p = 0.026. c Skin sensitization induced an inflammatory response, while novel formulation blocked inflammation onset. DNFB sensitization triggered upregulation of the inflammatory marker TNFa, but formulation B prevented inflammation onset. Two typical Western blots are shown. GAPDH, house-keeping protein. D, ACD; MAM + CGDS 2% (formulation A); MAM + CGDS 5% (formulation B). TNFa means + SEM, n = 7. *p = 0.249, #p = 0.210