Improvement effect of Xuebijing on brain tissue injury and Th17/Treg immune imbalance in cerebrospinal fluid in NMDA receptor encephalitis model mice

Lin CHEN,Limin YAN,Huaijie XING,Min CHEN,Xiaoyan LI,Chaosheng ZENG

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J Jilin Univ Med Ed ›› 2024, Vol. 50 ›› Issue (3) : 697-707. DOI: 10.13481/j.1671-587X.20240314
Research in basic medicine

Improvement effect of Xuebijing on brain tissue injury and Th17/Treg immune imbalance in cerebrospinal fluid in NMDA receptor encephalitis model mice

  • Lin CHEN,Limin YAN,Huaijie XING,Min CHEN,Xiaoyan LI,Chaosheng ZENG()
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Abstract

Objective To discuss the effect of Xuebijing on brain tissue damage and immune imbalance of helper T lymphocyte 17 (Th17)/regulatory T lymphocyte (Treg) in cerebrospinal fluid (CSF) of the N-methyl-D-aspartate (NMDA) receptor encephalitis model mice, and to clarify its therapeutic effect. Methods Sixty healthy male C57BL/6J mice were randomly divided into control group, model group, low dose of Xuebijing group, and high dose of Xuebijing group, and there were 15 mice in each group. Except for control group, the mice in the other three groups were injected with the antigen combined with immunostimulation to establish the NMDA receptor encephalitis models. The mice in low and high doses of Xuebijing groups were injected intraperitoneally with 5 and 10 mL·kg-1 of Xuebijing injection, respectively. HE staining was used to observe the pathomorphology of brain tissue of the mice in various groups; TUNEL assay was used to detect the apoptotic rates of the neurons in hippocampus CA1 region of brain tissue of the mice in various groups;enzyme-linked immunosorbent assay (ELISA) method was used to detect the levels of interleukin (IL)-6, IL-10, IL-17, and transforming growth factor β (TGF-β) in serum of the mice in various groups; flow cytometry was used to detect the percentages of Th17 and Treg cells in CSF of the mice in various groups; Western blotting method was used to detect the expression levels of retinoic acid-related orphan receptor γt (RORγt), forkhead box protein 3 (Foxp3), IL-10, and IL-17 proteins in brain tissue of the mice in various groups;immunohistochemistry method was used to detect the rates of IL-17 and Foxp3 positive cells in brain tissue of the mice in various groups. Results The HE staining results showed that the hippocampus CA1 region of brain tissue of the mice in control group had a clear structure without obvious lesions; compared with control group, the mice in model group showed partial pyramidal cell shrinkage, elongation of apical dendrites, loss of a few neurons, and sparse tissue in the hippocampus CA1 region of brain tissue; compared with model group, the mice in low and high doses of Xuebijing groups showed that the damage of the cells in the hippocampus CA1 region of brain tissue was decreased, and the morphological recovery, more orderly arrangement, and more significant improvement could be seen in hippocampus CA1 region of the mice in high dose of Xuebijing group. The TUNEL assay results showed that compared with control group, the apoptotic rate of the neurons in hippocampus CA1 region of brain tissue of the mice in model group was significantly increased (P<0.05); compared with model group, the apoptotic rate of the neurons in hippocampus CA1 region of brain tissue of the mice in low and high doses of Xuebijing groups were significantly decreased (P<0.05); compared with low dose of Xuebijing group, the apoptotic rate of the neurons in hippocampus CA1 region of brain tissue of the mice in high dose of Xuebijing group was significantly decreased (P<0.05). The ELISA results showed that compared with control group, the levels of IL-6 and IL-17 in serum of the mice in model group were significantly increased (P<0.05), while the levels of IL-10 and TGF-β were significantly decreased (P<0.05); compared with model group, the levels of IL-6 and IL-17 in serum of the mice in low and high doses of Xuebijing groups were significantly decreased (P<0.05), while the levels of IL-10 and TGF-β were significantly increased (P<0.05); compared with low dose of Xuebijing group, the levels of IL-6 and IL-17 in serum of the mice in high dose of Xuebijing group were significantly decreased (P<0.05), while the levels of IL-10 and TGF-β were significantly increased (P<0.05). The flow cytometry results showed that compared with control group, the percentage of CD4+IL-17A+ Th17 cells in CSF of the mice in model group was significantly increased (P<0.05), while the percentage of CD25+Foxp3+ Treg cells was significantly decreased (P<0.05); compared with model group, the percentages of CD4+IL-17A+ Th17 cells in CSF of the mice in low and high doses of Xuebijing groups were significantly decreased (P<0.05), while the percentage of CD25+Foxp3+ Treg cells was significantly increased (P<0.05); compared with low dose of Xuebijing group, the percentage of CD4+IL-17A+ Th17 cells in CSF of the mice in high dose of Xuebijing group was significantly decreased (P<0.05), while the percentage of CD25+Foxp3+ Treg cells was significantly increased (P<0.05). The Western blotting results showed that compared with control group, the expression levels of RORγt and IL-17 proteins in brain tissue of the mice in model group were significantly increased (P<0.05), while the expression levels of Foxp3 and IL-10 proteins were significantly decreased (P<0.05); compared with model group, the expression levels of RORγt and IL-17 proteins in brain tissue of the mice in low and high doses of Xuebijing groups were significantly decreased (P<0.05), while the expression levels of Foxp3 and IL-10 proteins were significantly increased (P<0.05); compared with low dose of Xuebijing group, the expression levels of RORγt and IL-17 proteins in brain tissue of the mice in high dose of Xuebijing group were significantly decreased (P<0.05), while the expression levels of Foxp3 and IL-10 proteins were significantly increased (P<0.05). The immunohistochemistry results showed that compared with control group, the rate of IL-17 positive cells in brain tissue of the mice in model group was significantly increased (P<0.05), while the rate of Foxp3 positive cells was significantly decreased (P<0.05); compared with model group, the rates of IL-17 positive cells in brain tissue of the mice in low and high doses of Xuebijing groups were significantly decreased (P<0.05), while the rates of Foxp3 positive cells were significantly increased (P<0.05); compared with low dose of Xuebijing group, the rate of IL-17 positive cells in brain tissue of the mice in high dose of Xuebijing group was significantly decreased (P<0.05), while the rate of Foxp3 positive cells was significantly increased (P<0.05). Conclusion Xuebijing can effectively ameliorate the brain tissue injury, regulate the cytokine levels, and intervene in immune imbalance of Th17/Treg in the mice with anti-NMDA receptor encephalitis.

Key words

Anti-N-methyl-D-aspartate receptor encephalitis / Xuebijing / Neuronal apoptosis / Helper T lymphocyte 17 / Regulatory T lymphocyte / Immune imbalance

CLC number

R512.3

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Lin CHEN,Limin YAN,Huaijie XING,Min CHEN,Xiaoyan LI,Chaosheng ZENG. Improvement effect of Xuebijing on brain tissue injury and Th17/Treg immune imbalance in cerebrospinal fluid in NMDA receptor encephalitis model mice. Journal of Jilin University(Medicine Edition). 2024, 50(3): 697-707 https://doi.org/10.13481/j.1671-587X.20240314

References

1 BASSAL F C, HARWOOD M, OH A, et al. Anti-NMDA receptor encephalitis and brain atrophy in children and adults: a quantitative study[J]. Clin Imaging, 2021, 78: 296-300.
2 GU J X, CHEN Q, GU H D, et al. Research progress in teratoma-associated anti-N-methyl-D-aspartate receptor encephalitis: the gynecological perspective[J]. J Obstet Gynaecol Res, 2021, 47(11): 3749-3757.
3 HUANG Q Y, XIE Y, HU Z P, et al. Anti-N-methyl-D-aspartate receptor encephalitis: a review of pathogenic mechanisms, treatment, prognosis[J]. Brain Res, 2020, 1727: 146549.
4 WALKER C A, POULIK J, D’MELLO R J. Anti-NMDA receptor encephalitis in an adolescent with a cryptic ovarian teratoma[J]. BMJ Case Rep, 2021, 14(7): e236340.
5 RAPHAEL I, JOERN R R, FORSTHUBER T G. Memory CD4+ T cells in immunity and autoimmune diseases[J]. Cells, 2020, 9(3): 531.
6 李红岩, 侯振江, 刘建凤, 等. Th17/Treg细胞及其细胞因子在神经系统自身免疫性疾病中的研究进展[J]. 医学综述, 2021, 27(5): 889-895, 900.
7 LEE G R. The balance of Th17 versus Treg cells in autoimmunity[J]. Int J Mol Sci, 2018, 19(3): 730.
8 朱 莉, 姜远安, 肖冠华, 等. MiR-125b在病毒性脑炎患儿中的表达水平及其与Th17/Treg平衡及预后的关系[J]. 疑难病杂志, 2021, 20(2): 129-133.
9 李维燕, 张丽平, 董俊刚. 血必净的临床应用研究进展[J]. 实用中医内科杂志, 2022, 36(10): 66-70.
10 CHEN X, FENG Y X, SHEN X Y, et al. Anti-sepsis protection of Xuebijing injection is mediated by differential regulation of pro- and anti-inflammatory Th17 and T regulatory cells in a murine model of polymicrobial sepsis[J]. J Ethnopharmacol, 2018, 211: 358-365.
11 叶 霖, 黄虹蜜, 吴 莹, 等. 抗NMDA受体脑炎相关癫痫小鼠脑组织小胶质细胞及IL-1β、TNF-α表达的变化[J]. 癫痫与神经电生理学杂志, 2022, 31(3): 129-134.
12 CHEN Z Q, ZHOU J T, WU D C, et al. Altered executive control network connectivity in anti-NMDA receptor encephalitis[J]. Ann Clin Transl Neurol, 2022, 9(1): 30-40.
13 WANG B S, ZOU L, ZHOU L. Lipid bilayers regulate allosteric signal of NMDA receptor GluN1 C-terminal domain[J]. Biochem Biophys Res Commun, 2021, 585: 15-21.
14 LIN K L, LIN J J. Neurocritical care for Anti-NMDA receptor encephalitis[J]. Biomed J, 2020, 43(3): 251-258.
15 NOSADINI M, EYRE M, MOLTENI E, et al. Use and safety of immunotherapeutic management of N-methyl-d-aspartate receptor antibody encephalitis: a meta-analysis[J]. JAMA Neurol, 2021, 78(11): 1333-1344.
16 CHEN Y, TONG H S, PAN Z G, et al. Xuebijing injection attenuates pulmonary injury by reducing oxidative stress and proinflammatory damage in rats with heat stroke[J]. Exp Ther Med, 2017, 13(6): 3408-3416.
17 LIU J F, WANG Z Z, LIN J, et al. Xuebijing injection in septic rats mitigates kidney injury, reduces cortical microcirculatory disorders, and suppresses activation of local inflammation[J]. J Ethnopharmacol, 2021, 276: 114199.
18 韩 睿, 李清廉, 刘 锐, 等. 血必净注射液对大鼠缺血性脑损伤的神经保护作用部分通过PI3K/Akt信号通路介导[J]. 中国实验诊断学, 2019, 23(2): 293-297.
19 张美琦, 王浩嘉, 李艺颖, 等. 血必净注射液对多种病毒的抑制作用及机制研究[J]. 药物评价研究, 2022, 45(9): 1697-1705.
20 YANG P, QIAN F Y, ZHANG M F, et al. Th17 cell pathogenicity and plasticity in rheumatoid arthritis[J]. J Leukoc Biol, 2019, 106(6): 1233-1240.
21 YASUDA K, TAKEUCHI Y, HIROTA K. The pathogenicity of Th17 cells in autoimmune diseases[J]. Semin Immunopathol, 2019, 41(3): 283-297.
22 OHKURA N, SAKAGUCHI S. Transcriptional and epigenetic basis of Treg cell development and function: its genetic anomalies or variations in autoimmune diseases[J]. Cell Res, 2020, 30(6): 465-474.
23 KUMAR R, THEISS A L, VENUPRASAD K. RORγt protein modifications and IL-17-mediated inflammation[J]. Trends Immunol, 2021, 42(11): 1037-1050.
24 ONO M. Control of regulatory T-cell differentiation and function by T-cell receptor signalling and Foxp3 transcription factor complexes[J]. Immunology, 2020, 160(1): 24-37.

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