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INNOSC Theranostics and
Pharmacological Sciences Antioxidant effects of curcumin in SCI

injection consisting of 80 mg/kg of ketamine and 15 mg/kg contain biochemical markers, were carefully gathered for
of xylazine. This combination was selected for its efficacy subsequent assays. To preserve the integrity of the samples,
in providing both sedation and analgesia, ensuring both tissue and blood supernatants were stored at –80°C.
the animals remained unconscious and free from pain
throughout the surgical procedure. Following anesthesia, 2.6. Measurement of OS
the fur over the surgical area was shaved to ensure a The present study focused on several key parameters,
sterile environment, minimizing the risk of infection. including malondialdehyde (MDA), GSH, SOD, and
A laminectomy was then performed at the T8 – T9 total antioxidant capacity (TAC). These parameters were
vertebral levels using a precision tool known as a micro measured using specific kits from ZellBio (Germany), and
rongeur (Fine Science Tools, USA). This step involved quantitation of OS and antioxidant levels in biological
removing a section of the vertebral bone to access the samples was conducted according to the manufacturer’s
spinal cord directly. To induce SCI, an extradural clip was protocols. The MDA assay (CAT No. ZB-MDA-96A) was
employed for compression, requiring the placement of an used to assess lipid peroxidation, indicating oxidative
aneurysm clip on the right side of the spinal cord. This clip damage. The GSH assay (CAT No. ZB-GSH-96A) measures
exerted a controlled closing force of 0.90 N for a duration the levels of this critical antioxidant, whereas the SOD assay
of 1 min, simulating the impact of a traumatic injury. After (CAT No. ZB-SOD-96A) evaluates the activity of this enzyme
the surgical procedure was completed, the incision site was that protects against OS. Finally, the TAC assay (CAT No.
meticulously suture-closed to promote healing and reduce ZB-TAC-96A) provides a comprehensive measure of the
the risk of complications during the recovery phase. This overall antioxidant capacity of the samples. Following the
careful approach ensures both the integrity of the surgical manufacturer’s instructions for each assay ensured accurate
site and the welfare of the animals involved in the study. and reproducible results, contributing to the understanding
of the biochemical changes associated with SCI and the
2.4. Motor behavior analysis potential therapeutic effects of the treatments administered.
The progression of functional recovery in each animal 2.7. Enzyme-linked immunosorbent assays (ELISA)
was meticulously evaluated using the Basso, Beattie, and
Bresnahan (BBB) test, a widely recognized and validated To measure the amounts of NLRP3, apoptosis-associated
behavioral assessment tool for measuring locomotor speck-like protein containing a CARD (ASC), and
function following SCI in rodent models. The BBB test caspase 1 (Casp-1) proteins, ELISA assays were employed
scores range from 0 to 21, with higher scores indicating to analyze freshly extracted spinal cord tissue samples.
better locomotor function. Each rat was observed for Upon collection, the spinal cord tissues were carefully
specific behaviors such as movement, coordination, and homogenized in a suitable lysis buffer to extract the
weight-bearing ability during a series of standardized tasks, proteins. Following homogenization, the samples were
which included walking on a flat surface and navigating centrifuged to remove cellular debris, leaving a clear
obstacles. Assessments began 24 h post-SCI and continued supernatant that contained the soluble proteins for
at 48 h and 72 h, with additional evaluations conducted analysis. ELISA kits for NLRP3 (MyBiosource, CA, United
weekly throughout the duration of the study (4 weeks). States), ASC (MyBiosource, CA, United States), and Casp-1
(Cusabio, Wuhan, China) that are specific for rats were
2.5. Sample preparation used to measure the protein levels of these three proteins
according to the manufacturers’ protocol.
After the completion of the experimental procedures
(after 4 weeks), the animals were humanely sacrificed in 2.8. Statistical analysis
accordance with ethical guidelines to minimize suffering. We used IBM Statistical Package for the Social Sciences
®
This process typically involved administering an overdose of Statistics software (ver. 20) to analyze data, and the results
anesthetic, ensuring that the animals were unconscious and are expressed as mean ± standard deviation. A one-way
pain-free before euthanasia. Following sacrifice, spinal cord analysis of variance was conducted followed by Tukey’s
tissue and blood samples were collected for further analysis. post hoc test to calculate the differences between groups.
The spinal cord tissues were carefully excised and stored in A P < 0.05 was considered to indicate statistical significance.
sterile containers, whereas blood samples were obtained
through cardiac puncture, a method that allows for the 3. Results
collection of blood directly from the heart, ensuring high-
quality samples. Once collected, the blood samples were 3.1. Functional recovery
centrifuged at 6000 rpm for 20 min to separate the plasma Our results showed that animals with SCI had significantly
from the cellular components. The supernatants, which lower BBB scores relative to the Control group animals

Volume 8 Issue 2 (2025) 79 doi: 10.36922/itps.4795