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Anti-inflammatory and Neuroprotective Effects of Transcranial Ultrasound Stimulation on Parkinson's Disease

Wen-Shin Song, Chen-Yu Sung, Chia-Hua Ke, Feng-Yi Yang

Ultrasound in Medicine & Biology 2022, 48, 265-274 · 10.1016/j.ultrasmedbio.2021.10.001

rodentparkinsons diseasehistology molecular

Abstract

Low-intensity pulsed ultrasound (LIPUS) is a promising non-invasive neuromodulation tool for deep brain stimulation. Here, we investigated the impact of LIPUS, including neuroprotective effects, on the pathology of Parkinson's disease (PD) in an animal model. Sprague-Dawley rats were injected with 6-hydroxydopamine (6-OHDA) at two sites in the right striatum. LIPUS (1 MHz, 5% duty cycle, 1-Hz pulse repetition frequency, 15 min/d) stimulation was then applied to some of the rats (the 6-OHDA + LIPUS group) beginning 2 wk after the 6-OHDA administration, while the remaining rats (the 6-OHDA group) received no LIPUS stimulation. The 6-OHDA-induced inflammatory responses and expressions of neurotrophic factors were quantified with immunofluorescence activity. The safety of LIPUS was assessed using hematoxylin and eosin and Nissl staining. LIPUS treatment significantly inhibited 6-OHDA-induced glial activation and the phosphorylation of nuclear factor-κB p65 in the substantia nigra pars compacta. Further study revealed that LIPUS effectively preserved the levels of neurotrophic factors, dopamine transporter and tight junction proteins of the blood-brain barrier in the 6-OHDA + LIPUS group compared with the 6-OHDA group. These results indicate that LIPUS acts via multiple neuroprotective mechanisms in the PD rat model and suggest that LIPUS can be viewed as a potential treatment for PD.

Abstract via europepmc.

Speciesrat (Sprague-Dawley)
Subjects4 animals
Sessions per subject30
Randomisedyes
Blindingsingle
Sham / controlnone
Auditory controlnot reported
Readout timingoffline
Anaesthesianot reported
Readoutshistology molecularimmunofluorescence (GFAP, Iba-1, IL-1β, phospho-NF-κB p65, GDNF, BDNF, DAT-TH double-label, ZO-1, claudin-5); hematoxylin & eosin and Nissl staining for safety
Direction of effectnot assessedLIPUS reduced 6-OHDA-induced glial activation and NF-kB/IL-1b signaling and restored GDNF, DAT, and tight-junction protein levels in the SNpc; this reflects anti-inflammatory/neuroprotective molecular outcomes rather than a measured direction of neuromodulatory (excitatory/inhibitory) effect.
Adverse eventsnone observedThe rats in the 6-OHDA + LIPUS treatment group did not exhibit changes in weight gain compared with the 6-OHDA group. H&E staining did not reveal any tissue damage or hemorrhaging associated with 5 d of LIPUS administration, while Nissl staining revealed that neuronal density appeared to be normal and indicated no neuronal damage throughout the entire brains.

Exposures

Exposure 1: LIPUS to lesioned striatal region in 6-OHDA rat model of Parkinson's disease

Target: striatum — “the targeted region in the lesions of the brain (right striatum, 6-OHDA injection site)
Device: Olympus / Panametrics · Panametrics · A392S

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)1,000✓✓
Pulse duration (ms)50✓✓
Pulse repetition frequency (Hz)1✓✓
Duty cycle (%)5pulse duration × PRF gives 5%✓✓
Sonication duration (s)300✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)0.528✓✓
In-situ estimatenot reported
In-situ pressure (kPa)not reported
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Protocol, in the paper’s words

The sonication was precisely targeted using a stereotaxic apparatus. The acoustic wave was delivered to the targeted region in the lesions of the brain. Each rat's right hemisphere was treated with LIPUS with triple sonications. The duration of each sonication was 5 min, and there was an interval of 5 min between sonications. LIPUS was applied consecutively to the rats in the 6-OHDA + LIPUS group 5 d/wk for 6 wk.

Flags from extraction

  • n_subjectsThe 4 6-OHDA+LIPUS rats are the group with clear reported LIPUS exposure; the paper also mentions unspecified-number additional healthy rats used for H&E/Nissl safety evaluation ('healthy rats were selected for safety evaluation... over 5 d in 1 wk') without stating their group size, so they are not included in the total.
  • n_sessions_per_subjectComputed as 3 sonications/day x 5 d/wk x 6 wk = 90 sonications, or 30 daily sessions; recorded as 30 (daily sessions) based on '5 d/wk for 6 wk' schedule.
  • anaesthesiaPaper states rats were anesthetized with isoflurane for the 6-OHDA stereotaxic injections but does not explicitly state whether the animals were anesthetized during the LIPUS sonication sessions themselves.
  • direction_of_effectStudy reports anti-inflammatory/neuroprotective histological outcomes, not a measured direction of neuromodulatory (excitatory/inhibitory) neural effect, so 'not_assessed' was used.
  • exposures[0].free_field.ispta_w_cm2Measured via radiation force balance in degassed water (no skull/tissue path stated for this measurement), assigned to free_field domain.