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,et al. Repairing Effects of Ginsenoside Rg1 on Traumatic Brain Injury in Mice[J]. Laboratory Animal and Comparative Medicine, 2023, 43(3): 243-252. DOI: 10.12300/j.issn.1674-5817.2022.187.

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.

References 26

1	LANGLOIS J A, RUTLAND-BROWN W, WALD M M. The epidemiology and impact of traumatic brain injury: a brief overview[J]. J Head Trauma Rehabil, 2006, 21(5):375-378. DOI: 10.1097/00001199-200609000-00001 .
2	KATZ D I, BERNICK C, DODICK D W, et al. National institute of neurological disorders and stroke consensus diagnostic criteria for traumatic encephalopathy syndrome[J]. Neurology, 2021, 96(18):848-863. DOI: 10.1212/WNL. 0000000000011850 .
3	SILVERBERG N D, IACCARINO M A, PANENKA W J, et al. Management of concussion and mild traumatic brain injury: a synthesis of practice guidelines[J]. Arch Phys Med Rehabil, 2020, 101(2):382-393. DOI: 10.1016/j.apmr.2019.10.179 .
4	GALGANO M, TOSHKEZI G, QIU X C, et al. Traumatic brain injury: current treatment strategies and future endeavors[J]. Cell Transplant, 2017, 26(7):1118-1130. DOI: 10.1177/0963689717714102 .
5	AHMED T, RAZA S H, MARYAM A, et al. Ginsenoside Rb1 as a neuroprotective agent: a review[J]. Brain Res Bull, 2016, 125:30-43. DOI: 10.1016/j.brainresbull.2016.04.002 .
6	GAO J, BAI H J, LI Q, et al. In vitro investigation of the mechanism underlying the effect of ginsenoside on the proliferation and differentiation of neural stem cells subjected to oxygen-glucose deprivation/reperfusion[J]. Int J Mol Med, 2018, 41(1):353-363. DOI: 10.3892/ijmm.2017.3253 .
7	WU Y X, WU H J, ZENG J X, et al. Mild traumatic brain injury induces microvascular injury and accelerates Alzheimer-like pathogenesis in mice[J]. Acta Neuropathol Commun, 2021, 9(1):74. DOI: 10.1186/s40478-021-01178-7 .
8	CUI W X, WU X, FENG D Y, et al. Acrolein induces systemic coagulopathy via autophagy-dependent secretion of von willebrand factor in mice after traumatic brain injury[J]. Neurosci Bull, 2021, 37(8):1160-1175. DOI: 10.1007/s12264-021-00681-0 .
9	CHEN Y F, LI J, MA B T, et al. MSC-derived exosomes promote recovery from traumatic brain injury via microglia/macrophages in rat[J]. Aging (Albany NY), 2020, 12(18):18274-18296. DOI: 10.18632/aging.103692 .
10	LONG X B, YAO X L, JIANG Q, et al. Astrocyte-derived exosomes enriched with miR-873a-5p inhibit neuroinflammation via microglia phenotype modulation after traumatic brain injury[J]. J Neuroinflammation, 2020, 17(1):89. DOI: 10.1186/s12974-020-01761-0 .
11	ZHANG Y L, ZHANG Y, CHOPP M, et al. Mesenchymal stem cell-derived exosomes improve functional recovery in rats after traumatic brain injury: a dose-response and therapeutic window study[J]. Neurorehabil Neural Repair, 2020, 34(7):616-626. DOI: 10.1177/1545968320926164 .
12	ARAKI T, YOKOTA H, MORITA A. Pediatric traumatic brain injury: characteristic features, diagnosis, and management[J]. Neurol Med Chir (Tokyo), 2017, 57(2):82-93. DOI: 10.2176/nmc.ra.2016-0191 .
13	雷勋明. 人参皂苷Rg1对新生鼠缺氧缺血性脑损伤海马神经元凋亡及学习记忆能力的影响[J]. 中国中西医结合儿科学, 2018, 10(4): 277-279. DOI: 10.3969/j.issn.1674-3865.2018.04.001 .
	LEI X M. Effects of ginseng Rg1 on hippocampal neuronal apoptosis and learning ability of neonatal rats with hypoxic-ischemic brain damage[J]. Chin Pediatr Integr Tradit West Med, 2018, 10(4): 277-279. DOI: 10.3969/j.issn.1674-3865.2018.04.001 .
14	SONG X Y, HU J F, CHU S F, et al. Ginsenoside Rg1 attenuates okadaic acid induced spatial memory impairment by the GSK3β/tau signaling pathway and the Aβ formation prevention in rats[J]. Eur J Pharmacol, 2013, 710(1-3):29-38. DOI: 10.1016/j.ejphar.2013.03.051 .
15	ZOU S F, CHEN W, DING H, et al. Involvement of autophagy in the protective effects of ginsenoside Rb1 in a rat model of traumatic brain injury[J]. Eur J Drug Metab Pharmacokinet, 2022, 47(6):869-877. DOI: 10.1007/s13318-022-00799-0 .
16	ZHANG Z R, SONG Z J, SHEN F M, et al. Ginsenoside Rg1 prevents PTSD-like behaviors in mice through promoting synaptic proteins, reducing Kir4.1 and TNF-α in the Hippocampus[J]. Mol Neurobiol, 2021, 58(4):1550-1563. DOI: 10.1007/s12035-020-02213-9 .
17	HU B Y, LIU X J, QIANG R, et al. Treatment with ginseng total saponins improves the neurorestoration of rat after traumatic brain injury[J]. J Ethnopharmacol, 2014, 155(2):1243-1255. DOI: 10.1016/j.jep.2014.07.009 .
18	ZHAI K F, DUAN H, WANG W, et al. Ginsenoside Rg1 ameliorates blood-brain barrier disruption and traumatic brain injury via attenuating macrophages derived exosomes miR-21 release[J]. Acta Pharm Sin B, 2021, 11(11):3493-3507. DOI: 10.1016/j.apsb.2021.03.032 .
19	DHANDA S, SANDHIR R. Blood-brain barrier permeability is exacerbated in experimental model of hepatic encephalopathy via MMP-9 activation and downregulation of tight junction proteins[J]. Mol Neurobiol, 2018, 55(5):3642-3659. DOI: 10.1007/s12035-017-0521-7 .
20	REMPE R G, HARTZ A M S, BAUER B. Matrix metallo-proteinases in the brain and blood-brain barrier: versatile breakers and makers[J]. J Cereb Blood Flow Metab, 2016, 36(9):1481-1507. DOI: 10.1177/0271678X16655551 .
21	LI Y H, MENG Q, YANG M B, et al. Current trends in drug metabolism and pharmacokinetics[J]. Acta Pharm Sin B, 2019, 9(6):1113-1144. DOI: 10.1016/j.apsb.2019.10.001 .
22	XIONG Y, MAHMOOD A, CHOPP M. Animal models of traumatic brain injury[J]. Nat Rev Neurosci, 2013, 14(2):128-142. DOI: 10.1038/nrn3407 .
23	KARVE I P, TAYLOR J M, CRACK P J. The contribution of astrocytes and microglia to traumatic brain injury[J]. Br J Pharmacol, 2016, 173(4):692-702. DOI: 10.1111/bph.13125 .
24	SOFRONIEW M V, VINTERS H V. Astrocytes: biology and pathology[J]. Acta Neuropathol, 2010, 119(1):7-35. DOI: 10.1007/s00401-009-0619-8 .
25	VILLAPOL S, BYRNES K R, SYMES A J. Temporal dynamics of cerebral blood flow, cortical damage, apoptosis, astrocyte-vasculature interaction and astrogliosis in the pericontusional region after traumatic brain injury[J]. Front Neurol, 2014, 5:82. DOI: 10.3389/fneur.2014.00082 .
26	KOLIATSOS V E, RAO V. The behavioral neuroscience of traumatic brain injury[J]. Psychiatr Clin North Am, 2020, 43(2):305-330. DOI: 10.1016/j.psc.2020.02.009 .

[1]	LIU Yueqin, XUE Weiguo, WANG Shuyou, SHEN Yaohua, JIA Shuyong, WANG Guangjun, SONG Xiaojing. Observation of Digestive Tract Tissue Morphology in Mice Using Probe-Based Confocal Laser Endomicroscopy [J]. Laboratory Animal and Comparative Medicine, 2025, 45(4): 457-465.
[2]	KONG Zhihao, WEI Xiaofeng, YU Lingzhi, FENG Liping, ZHU Qi, SHI Guojun, WANG Chen. Isolation and Identification of Staphylococcus xylosus in Nude Mice with Squamous Skin Scurfs [J]. Laboratory Animal and Comparative Medicine, 2025, 45(3): 368-375.
[3]	XU Qiuyu, YAN Guofeng, FU Li, FAN Wenhua, ZHOU Jing, ZHU Lian, QIU Shuwen, ZHANG Jie, WU Ling. A Mouse Model of Polycystic Ovary Syndrome Established Through Subcutaneous Administration of Letrozole Sustained-Release Pellets and Hepatic Transcriptome Analysis [J]. Laboratory Animal and Comparative Medicine, 2025, 45(2): 119-129.
[4]	LIU Rongle, CHENG Hao, SHANG Fusheng, CHANG Shufu, XU Ping. Study on Cardiac Aging Phenotypes of SHJH ^hr Mice [J]. Laboratory Animal and Comparative Medicine, 2025, 45(1): 13-20.
[5]	WU Zhihao, CAO Shuyang, ZHOU Zhengyu. Establishment of an Intestinal Fibrosis Model Associated with Inflammatory Bowel Disease in VDR^-/- Mice Induced by Helicobacter hepaticus Infection and Mechanism Exploration [J]. Laboratory Animal and Comparative Medicine, 2025, 45(1): 37-46.
[6]	ZHANG Nan, LI Huaiyin, LIAN Xiaodi, WEI Juanpeng, GAO Ming. Effects of Different Durations of Light Exposure on Body Weight and Learning and Memory Abilities of NIH Mice [J]. Laboratory Animal and Comparative Medicine, 2025, 45(1): 73-78.
[7]	ZHAO Xiaona, WANG Peng, YE Maoqing, QU Xinkai. Establishment of a New Hyperglycemic Obesity Cardiac Dysfunction Mouse Model with Triacsin C [J]. Laboratory Animal and Comparative Medicine, 2024, 44(6): 605-612.
[8]	TAN He, YANG Xiaohui, ZHANG Daxiu, WANG Guicheng. Optimal Adaptation Period for Metabolic Cage Experiments in Mice at Different Developmental Stages [J]. Laboratory Animal and Comparative Medicine, 2024, 44(5): 502-510.
[9]	MENG Yu, LIANG Dongli, ZHENG Linlin, ZHOU Yuanyuan, WANG Zhaoxia. Optimization and Evaluation of Conditions for Orthotopic Nude Mouse Models of Human Liver Tumor Cells [J]. Laboratory Animal and Comparative Medicine, 2024, 44(5): 511-522.
[10]	Jing QIN, Yong ZHAO, Caiqin ZHANG, Bing BAI, Changhong SHI. Construction and Evaluation of Theranostic Near-infrared Fluorescent Probe for Targeting Inflammatory Brain Edema [J]. Laboratory Animal and Comparative Medicine, 2024, 44(3): 243-250.
[11]	Yisu ZHANG, Xinru LIU, Ruojie WU, Rui LIU, Hong OUYANG, Xiaohong LI. Establishment and Evaluation of Mouse Model of Pregnancy Pain-depression Comorbidity Induced by Chronic Unpredictable Stress, Complete Freund's Adjuvant and Formalin [J]. Laboratory Animal and Comparative Medicine, 2024, 44(3): 259-269.
[12]	Dong WU, Rui SHI, Peishan LUO, Ling'en LI, Xijing SHENG, Mengyang WANG, Lu NI, Sujuan WANG, Huixin YANG, Jing ZHAO. Effects of Different Pellet Feed Hardness on Growth and Reproduction, Feed Utilization Rate, and Environmental Dust in Laboratory Mice [J]. Laboratory Animal and Comparative Medicine, 2024, 44(3): 313-320.
[13]	Yun LIU, Tingting FENG, Wei TONG, Zhi GUO, Xia LI, Qi KONG, Zhiguang XIANG. Glycyrrhizic Acid Showed Therapeutic Effects on Severe Pulmonary Damages in Mice Induced by Pneumonia Virus of Mice Infection [J]. Laboratory Animal and Comparative Medicine, 2024, 44(3): 251-258.
[14]	Jinhua HU, Jingjie HAN, Min JIN, Bin HU, Yuefen LOU. Effects of Puerarin on Bone Density in Rats and Mice: A Meta-analysis [J]. Laboratory Animal and Comparative Medicine, 2024, 44(2): 149-161.
[15]	Min LIANG, Yang GUO, Jinjin WANG, Mengyan ZHU, Jun CHI, Yanjuan CHEN, Chengji WANG, Zhilan YU, Ruling SHEN. Construction of Dmd Gene Mutant Mice and Phenotype Verification in Muscle and Immune Systems [J]. Laboratory Animal and Comparative Medicine, 2024, 44(1): 42-51.