Investig Clin Urol.  2019 Sep;60(5):359-366. 10.4111/icu.2019.60.5.359.

Treatment with low-energy shock wave alleviates pain in an animal model of uroplakin 3A-induced autoimmune interstitial cystitis/painful bladder syndrome

• Li H
• Zhang Z
• Peng J
• Xin Z
• Li M
• Yang B
• Fang D
• Tang Y
• Guo Y

• KMID: 2455963
• DOI: http://doi.org/10.4111/icu.2019.60.5.359

Abstract

PURPOSE
To investigate whether treatment with low-energy shock wave (LESW) alleviates pain and bladder dysfunction in a mouse model of uroplakin 3A (UPK3A)-induced interstitial cystitis/painful bladder syndrome (IC/PBS).
MATERIALS AND METHODS
Forty female BALB/c mice were divided into four groups (n=10/group): Sham, Sham+LESW, UPK3A, and UPK3A+LESW. At 6 weeks of age, mice were injected with an emulsion containing water and complete Freund's adjuvant with (UPK3A and UPK3A+LESW groups) or without (Sham and Sham+LESW groups) 200 µg of UPK3A. At 10 weeks, mice received a second dose of Freund's adjuvant to booster immunization. At 12 weeks, mice underwent pain assessment and a frequency volume chart (FVC) test as the pretreatment assessment. LESW treatment and pain assessment were conducted from 13 to 15 weeks. One week after the final treatment, pain assessment and the FVC were conducted again as the post-treatment assessment. Mice were euthanized and sacrificed at 17 weeks.
RESULTS
The presence of tactile allodynia and bladder dysfunction was significant in the UPK3A-injected mice. LESW raised the pain threshold and improved bladder function with decreased urinary frequency and increased mean urine output. Expression and secretion of local and systemic inflammatory markers, including tumor necrosis factor-α (TNF-α) and nerve growth factor (NGF), increased after UPK3A immunization. These markers were significantly decreased after LESW treatment (p<0.05).
CONCLUSIONS
LESW treatment attenuated pain and bladder dysfunction in a UPK3A-induced model of IC/PBS. Local and systemic inflammation was partially controlled, with a reduced number of infiltrated inflammatory cells and reduced levels of TNF-α and NGF.

Keyword

Cystitis, interstitial; Inflammation; Pain; Shockwave, ultrasonic

MeSH Terms

Animals*
Cystitis, Interstitial
Female
Freund's Adjuvant
High-Energy Shock Waves
Humans
Hyperalgesia
Immunization
Immunization, Secondary
Inflammation
Mice
Models, Animal*
Necrosis
Nerve Growth Factor
Pain Measurement
Pain Threshold
Shock*
Urinary Bladder*
Uroplakins*
Water
Freund's Adjuvant
Nerve Growth Factor
Uroplakins
Water

Figure

• 

  Fig. 1 Presence of tactile allodynia in experimental autoimmune cystitis (EAC) mice and pain-alleviating effect of low-energy shock wave (LESW) treatment. Mechanical stimulation with von Frey filaments of increasing size on the suprapubic region resulted in significantly greater tactile sensitivity in the EAC mouse model than in uninjected mice (A–E). Fifty percent threshold forces were significantly decreased after uroplakin 3A (UPK3A) immunization (F). LESW treatment of UPK3A-immunized mice significantly alleviated the local pain and increased 50% threshold forces (A–F). *p<0.05 compared to Sham. #p<0.05 compared to UPK3A.

• 

  Fig. 2 Low-energy shock wave (LESW) treatment decreases urinary frequency and increases mean urine output in an experimental autoimmune cystitis (EAC) mouse model. Mice immunized with uroplakin 3A (UPK3A) showed significantly increased micturition frequency and significantly decreased mean urine outputs (A, B). LESW treatment decreased the urinary frequency and increased the mean urine output in the UPK3A+LESW group compared with the UPK3A group (C, D). *p<0.05 compared to Sham. #p<0.05 compared to UPK3A.

• 

  Fig. 3 Hematoxylin & eosin staining (10× and 200×) to show bladder wall thickness and inflammatory cell infiltration influenced by uroplakin 3A (UPK3A) injection and low-energy shock wave (LESW) treatment.

• 

  Fig. 4 Systemic and local inflammation in bladder tissue was induced by uroplakin 3A (UPK3A) immunization, and low-energy shock wave (LESW) treatment decreased local inflammatory reactions. (A) Serum levels of tumor necrosis factor-α (TNF-α), nerve growth factor (NGF), interleukin (IL)-2, and IL-6, respectively, influenced by UPK3A immunization and LESW treatment. (B) Reverse transcription polymerase chain reaction examination of expression levels of NGF, interferon-γ (IFN-γ), TNF-α, and IL-2, -4, -6, and -8. (C) Western blot examination of protein levels of TNF-α, NGF, and downstream inflammatory marker nuclear factor-κB (NF-κB). (D) Calculation of Western blot results based on bands' grey scale. *p<0.05 compared to Sham. #p<0.05 compared to UPK3A.

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