Establishment of spastic brain injury model in rats
Zhou Tong1,2, Bu Wei3, Zhang Xu1, Yu Xiaofei1, Bai Yanbin1, Li Hongjie1, Cao Ran1,4, Yu Yadong1
1Department of Hand Surgery, The Third Hospital of Hebei Medical University, Shijiazhuang 050051, China; 2Department of Hand Surgery, The Second Hospital of Tangshan, Tangshan 063000, China; 3Department of Neurosurgery, The Third Hospital of Hebei Medical University, Shijiazhuang 050051, China; 4Department of Orthopaedics, Shijiazhuang People's Hospital, Shijiazhuang 050036, China
Abstract：Objective To explore a method of establishing a rat spastic brain injury model. Methods Sixty specified pathogen-free grade male SD rats aged 2–3 months, with a body mass of 290–320 (304.2±11.6) g, were divided into the ethanol group (ETH group, n=20), normal saline group (NS group, n=20), and blank control group (CK group, n=20) by random number method. According to the stereotaxic map of the rat brain, the location of the right internal capsule of the rat was determined by the brain stereotaxic instrument. Then, we injected 80 μL of ethanol into the right internal capsule in the ETH group. Analogously, 80 μL of normal saline and no reagent were injected into the same target in the NS group and CK group, respectively. The general conditions of the rats in the three groups were observed after operation. The onset time and duration of symptoms and the mortality were recorded. On the 3rd, 7th, 14th, and 21st day after operation, the degree of brain injury was evaluated by neurological severity scores (NSS). The state of motor injury was evaluated by Faden scores, and muscle spasm was evaluated by modified Ashworth scores. On the 21st day after operation, the brain tissues of rats in each group were taken to prepare sections, and HE staining was used to observe the injury of the internal capsule. The total flexor muscle of the left forelimb and the gastrocnemius muscle of the left hindlimb were taken to prepare sections, and the percentage of type Ⅰ muscle fibers was detected by immunofluorescence. Results Within 3 days after surgery, 5 rats in ETH group, 2 in NS group and 1 in CK group died. The rats in the ETH group showed spasm in the left upper and lower limbs one day after operation, and the symptoms lasted for 21 days. However, no spasm of the left upper and lower limbs was observed in the NS and CK group. The NSS and modified Ashworth scores were all 0 in the NS and CK groups on the 3rd, 7th, 14th, and 21st day after operation, while their Faden scores were 22 points. In the ETH group, their NSS scores were (15.5±1.9), (15.8±1.8), (15.4±1.7), and (15.1±1.8) points on the 3rd, 7th, 14th, and 21st day after operation, respectively. Their Faden scores were (3.5±1.8), (3.2±1.7), (3.7±1.9), and (3.9±1.8) points, while their modified Ashworth scores were (3.5±0.5), (3.5±0.5), (2.9±0.7), and (2.9±0.6) points, respectively. The brain morphology HE stain showed hollow necrosis in the right internal capsule, a significant decrease in nerve cells, loose and disordered structure of nerve cells, shrunk or absent nucleus, and no involvement of other brain regions. In the NS and CK groups, the structure of brain nerve cells was intact and orderly, with no distortion or deformation of the nucleus and nucleoli. Immunofluorescence detection showed that the percentage of type Ⅰ muscle fibers of the total flexor muscle of the forearm of the left limbs was 10.2%±6.3%, 6.5%±2.9%, and 6.7%±2.9% in the ETH, NS, and CK groups, respectively. The percentage of type Ⅰ muscle fibers of the total flexor muscle of the gastrocnemius muscle of the contralateral limbs was 13.8%±5.1%, 7.7%±3.3%, and 7.6%±4.8% in the ETH, NS, and CK groups, respectively. The ratio of type Ⅰ muscle fiber of the total flexor muscle of the upper limb and gastrocnemius muscle of the lower limb in the ETH group was significantly higher than that in the NS and CK groups. The differences were all statistically significant (all P values <0.05). Conclusion Targeted injection into the internal capsule with ethanol can establish a typical and stable model of spastic brain injury in adult SD rat.
周彤, 步玮, 张旭, 于晓飞, 白延彬, 李洪杰, 曹冉, 于亚东. 大鼠痉挛性脑损伤模型的建立[J]. 中华解剖与临床杂志, 2022, 27(3): 195-200.
Zhou Tong, Bu Wei, Zhang Xu, Yu Xiaofei, Bai Yanbin, Li Hongjie, Cao Ran, Yu Yadong. Establishment of spastic brain injury model in rats. Chinese Journal of Anatomy and Clinics, 2022, 27(3): 195-200.
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