Repairing Effects of Ginsenoside Rg1 on Traumatic Brain Injury in Mice

doi:10.12300/j.issn.1674-5817.2022.187

Abstract

Abstract:

Objective To explore the effects of ginsenoside Rg1 on blood-brain barrier, neuroinflammation and behavioral function of traumatic brain injury (TBI) mouse model. Methods The experiment was divided into two parts. In the first part, 27 SPF male BALB/c mice were randomly divided into blank group, sham operation group and TBI model group, with 9 mice in each group. TBI model group was made by controlled cortical impact (CCI) after craniotomy, while sham operation group was only performed craniotomy without any treatment, and the blank group was not treated at all. The effect of modeling was evaluated after operation. In the second part, 50 male BALB/c mice were randomly divided into sham operation group, three different drug dosage groups and solvent (DMSO) control group, with 8 mice in each group. The drug treatment groups were injected with ginsenoside Rg1 at the doses of 10, 20 and 40 mg/kg respectively 6 hours after TBI model had been successfully established, while the DMSO control group was given the same amount of 1% DMSO for one week, twice a day. Modified neurological severity scores (mNSS) were performed on the 1st, 3rd, 7th and 14th day after modeling, and the blood-brain barrier leakage was detected by Western blotting on the 3rd day after modeling. On the 14th and 16th day, the elevated cross maze test and water maze test were used to detect the neurobehavioral function. On the 28th day after anesthesia and perfusion, the brains were taken out, and the neuroinflammation such as activation of microglia and astrocytes was observed by immunofluorescence staining. Results The expression level of MMP-9, a marker of blood-brain barrier, decreased in ginsenoside Rg1 treatment group (P<0.01). The number of microglia （Iba-1 positive) and astrocyte (GFAP positive) cells decreased significantly (P<0.05), which indicated that neuroinflammation was inhibited, and the best effect was achieved at the dosage of 20 mg/kg (P<0.01). The mNSS of mice in ginsenoside Rg1 treatment group were significantly lower than those in DMSO control group (P < 0.01), and the proportion of times they entered the open arm was significantly higher than that in DMSO control group (P < 0.05). The time ratio in the quadrant where the water maze experimental platform was located and the times of crossing the platform were significantly higher than those in control group (P < 0.05), and the dosage of 20 mg/kg had the best effect. Conclusion The TBI mouse model was successfully constructed and applied to the study of ginsenoside Rg1 repair of mouse traumatic brain injury. Ginsenoside Rg1 can significantly improve blood-brain barrier, alleviate neuroinflammation and improve neurobehavioral function in TBI model mice, and the effect is the most significant at the dose of 20 mg/kg.

Key words: Traumatic brain injury, Ginsenoside Rg1, Mice, Nervoinflammation, Ethology

CLC Number:

R-332

Wenwen GUO, Ya ZHAO, Yinghua WANG, Ke LIU, Xu GE, Yanying ZHANG, Yongfeng WANG, Changhong SHI. Repairing Effects of Ginsenoside Rg1 on Traumatic Brain Injury in Mice[J]. Laboratory Animal and Comparative Medicine, 2023, 43(3): 243-252.

Figures/Tables 4

Figure 1 Establishment and evaluation of traumatic brain injury （TBI） model in miceNote： A, Observation of brain tissue injury (the white arrow refers to damaged brain region by TBI); B, Neurological function score (mNSS is the improved nerve injury severity score); C, Elevated cross maze experiment; D-E, Water maze experiment. Control is the blank group without any treatment; Sham is the sham operation group; TBI is TBI modeling group by controlled cortical impact method, with 9 mice in each group. Compared with the sham group, ?P<0.05, ??P＜0.01, ???P＜0.001; Compared with control group, #P<0.05, ##P<0.01.

Figure 2 Effect of ginsenoside Rg1 on blood-brain barrier after traumatic brain injury （TBI） in miceNote： MMP-9, matrix metalloprotein-9. Sham is the sham operation group; DMSO is the solvent control group after TBI modeling by controlled cortical impact method; 10 mg/kg, 20 mg/kg， and 40 mg/kg Rg1 are 10, 20, 40 mg/kg of ginsenoside Rg1 treatment groups after TBI modeling by controlled cortical impact method, with 8 mice in each group. Compared with DMSO group, ??P＜0.01.

Figure 3 Effect of ginsenoside Rg1 on microglia and astrocytes 28 d after traumatic brain injury （TBI） in miceNote： A-B show ionized calcium binding adapter molecule 1 (Iba-1） protein expression (reflecting microglia activity) and the number of Iba-1 positive cells in brain tissues of mice in each group detected by immunofluorescence staining. C-D show glial fibrillary acidic protein （GFAP） expression (reflecting the activity of astrocytes) and the number of GFAP positive cells in the brain tissue of mice in each group detected by immunofluorescence staining. In Figures A and C, the top, middle, and bottom three photos in each group show three slices of similar layers in the brain tissue of three mice. Sham is the sham operation group; DMSO is the solvent control group after TBI modeling by controlled cortical impact method; 10 mg/kg, 20 mg/kg, 40 mg/kg Rg1 are 10, 20, 40 mg/kg of ginsenoside Rg1 treatment groups after TBI modeling by controlled cortical impact method, with 8 mice in each group. Compared with DMSO group, ?P<0.05， ??P<0.01.

Figure 4 Effect of ginsenoside Rg1 on behavioral performance in mice after traumatic brain injury （TBI）Note： A， mNSS neurological function score; B, Elevated cross maze experiment; C-D, water maze experiment. Sham is the sham operation group; DMSO is the solvent control group after TBI modeling by controlled cortical impact method; 10 mg/kg, 20 mg/kg, 40 mg/kg Rg1 are 10, 20, 40 mg/kg of ginsenoside Rg1 treatment groups after TBI modeling by controlled cortical impact method, with 8 mice in each group. Compared with Sham group, #P<0.05, ##P<0.01; compared with DMSO group, ?P＜0.05， ??P＜0.01.

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