Ozone therapy ameliorates tubulointerstitial inflammation by regulating TLR4 in adenine-induced CKD rats

Ren Fail. 2016 Jun;38(5):822-30. doi: 10.3109/0886022X.2016.1143757. Epub 2016 Feb 24.


Tubulointerstitium inflammation is a common pathway aggravating chronic kidney disease (CKD) progression and the mechanism is partly associated with excessive activation of toll-like receptor 4 (TLR4) in tubulointerstitium. Ozone therapy is demonstrated to alleviate inflammation in some experiments. The aim of this study is to examine whether ozone therapy could ameliorate chronic tubulointerstitium inflammation by suppressing TLR4 in adenine-induced CKD rats. Sprague-Dawley rats were fed with 0.75% adenine-containing diet to induce CKD and tubulointerstitium inflammation injury. Ozone therapy (1.1 mg/kg) was simultaneously administrated by rectal insufflations (i.r.). After 4 weeks, serum and kidney samples were collected for detection. Renal function and systemic electrolyte were detected. Renal pathological changes were assessed by hematoxylin-eosin (H&E) staining and Masson trichrome (MT) staining. Immunohistochemistry, Western blot and Real-time PCR were applied to evaluate tubulointerstitium inflammation as well as the expression of TLR4 and phosphorylated nuclear factor kappa B P65 (p-NF-κB P65) in rats. The results showed ozone therapy improved serious renal insufficiency, systemic electrolyte disorder and tubulointerstitium morphology damages in adenine-induced CKD rats. In addition, ozone therapy suppressed excessive activation of TLR4 and p-NF-κB P65 in the tubulointerstitium of adenine-induced CKD rats, accompanied by the reduction of inflammation-related cytokines including monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6). The protein expression of TLR4 was positively correlated with the protein expression levels of MCP-1 (r = 0.7863, p < 0.01) and TNF-α (r = 0.7547, p < 0.01) in CKD rats. These findings indicated ozone therapy could attenuate tubulointerstitium inflammation injury in adenine-induced CKD rats and the mechanism might associate with the mediation of TLR4.

Keywords: Adenine; ozone therapy; rat; toll-like receptor 4; tubule-interstitial inflammation.

MeSH terms

  • Animals
  • Disease Models, Animal
  • Disease Progression
  • Immunohistochemistry
  • Inflammation / metabolism
  • Kidney / metabolism
  • Kidney / pathology
  • Kidney Function Tests / methods
  • NF-kappa B / metabolism
  • Oxidants, Photochemical / pharmacology
  • Ozone / pharmacology*
  • Rats
  • Rats, Sprague-Dawley
  • Renal Insufficiency, Chronic* / diagnosis
  • Renal Insufficiency, Chronic* / metabolism
  • Renal Insufficiency, Chronic* / therapy
  • Toll-Like Receptor 4 / metabolism*
  • Treatment Outcome


  • NF-kappa B
  • Oxidants, Photochemical
  • Tlr4 protein, rat
  • Toll-Like Receptor 4
  • Ozone