Abstract
目的
表没食子儿茶素没食子酸酯(epigallocatechin gallate，EGCG)是茶叶中茶多酚生物活性的主要成分，具有多种生物学效应。脑缺血再灌注(ischemia/reperfusion，I/R)导致的神经损伤与炎症反应密切相关，促炎细胞因子白细胞介素-1β(interleukin-1β，IL-1β)与抗炎细胞因子白细胞介素-10(interleukin-10，IL-10)在炎症反应过程中起重要的调节作用。本研究旨在探讨EGCG对脑I/R损伤大鼠脑组织含水量、髓过氧化物酶(myeloperoxidase，MPO)活性以及IL-1β、IL-10的影响。
方法
采用线栓法建立SD大鼠大脑中动脉栓塞模型，大鼠缺血1.5 h后，将尼龙线拔出，行再灌注24.0 h。将SD大鼠随机分成假手术(Sham)组、I/R组、EGCG1组(I/R+12.5 mg/kg EGCG)、EGCG2组 (I/R+25.0 mg/kg EGCG)、EGCG3组(I/R+50.0 mg/kg EGCG)5组。Sham组大鼠除不予栓塞大脑中动脉外，其余与I/R组相同。采用干湿重法检测各组大鼠脑组织含水量，比色法检测MPO活性，RT-PCR测定IL-1β及IL-10 mRNA水平，ELISA测定IL-1β和IL-10含量。另取原代培养的SD大鼠大脑皮层神经元随机分成对照(Control)组、氧糖剥夺/再灌注(oxygen-glucose deprivation/reperfusion，OGD/R)组、OGD/R+EGCG组(OGD/R+50.0 μmol/L EGCG)3组，采用蛋白质印迹法检测神经元IL-1β和IL-10蛋白质的表达水平。
结果
与sham组比较，I/R组大鼠脑组织含水量、MPO活性、IL-1β含量、IL-10含量均显著增加(P<0.05或P<0.01)，IL-1β及IL-10 mRNA表达水平均明显上调(均P<0.01)。与I/R组比较，EGCG3组大鼠脑组织含水量、MPO活性均显著降低(均P<0.01)，IL-1β含量显著减少，IL-10含量显著增加(均P<0.01)。与I/R组比较，EGCG2组、EGCG3组大鼠缺血区脑组织IL-1β mRNA水平显著下调、IL-10 mRNA水平显著上调(P<0.05或P<0.01)。与Control组比较，OGD/R组神经元IL-1β及IL-10蛋白质表达水平均显著上调(均P<0.01)。与OGD/R组比较，OGD/R+EGCG组神经元IL-1β蛋白质表达明显下调，IL-10蛋白质表达明显上调(均P<0.01)。
结论
EGCG可抑制脑缺血损伤引发的炎症反应，这一效应可能与抑制促炎细胞因子IL-1β和促进抗炎细胞因子IL-10的表达有关。
Keywords: 表没食子儿茶素没食子酸酯, 缺血再灌注, 炎症反应, 白细胞介素-1β, 白细胞介素-10
Abstract
Objective
Epigallocatechin gallate (EGCG) is the main bioactive component of polyphenols in tea, which has a variety of biological effects. The neurologic injury caused by cerebral ischemia/reperfusion (I/R) is closely related to the inflammatory reaction. The pro-inflammatory factor interleukin-1β (IL-1β) and the anti-inflammatory factor interleukin-10 (IL-10) play important roles in the regulation of inflammatory reaction. This study aims to explore the effect of EGCG on water content, myeloperoxidase (MPO) activity, IL-1β and IL-10 in brain tissues of rats suffered cerebral I/R injury.
Methods
Middle cerebral artery occlusion model of SD rats was established by suture embolic method. After ischemia for 1.5 h, the nylon thread was pulled out and reperfusion was performed for 24.0 h. SD rats were randomly divided into 5 groups: a sham group, an I/R group, a EGCG1 group (I/R+12.5 mg/kg EGCG), a EGCG2 (I/R+25.0 mg/kg EGCG), and a EGCG3 group (I/R+50.0 mg/kg EGCG). The sham group was the same as the I/R group except that the middle cerebral artery was not blocked. The water content in brain tissues of rats was measured by dry and wet weight method, and MPO activity was detected by colorimetry. The levels of IL-1β and IL-10 mRNA were determined by RT-PCR, and the contents of IL-1β and IL-10 were determined by ELISA. In addition, primary cultured cortical neurons of SD rats were randomly divided into 3 groups: a control group, an oxygen-glucose deprivation/reperfusion (OGD/R) group, and an OGD/R+EGCG group (OGD/R+50.0 μmol/L EGCG). The effects of EGCG on the levels of IL-1β and IL-10 protein in neurons were assessed by Western blotting.
Results
Compared with the sham group, water content, MPO activity, the contents of IL-1β and IL-10 were significantly increased (P<0.05 or P<0.01), the mRNA expression of IL-1β and IL-10 were obviously up-regulated (both P<0.01) in cerebral tissues of rats in the I/R group. Compared with the I/R group, water content and MPO activity in cerebral tissues of rats were significantly decreased (both P<0.01), the content of IL-1β (P<0.01) were significantly decreased and IL-10 (P<0.01) were significantly increased in the EGCG3 group. Compared with the I/R group, the mRNA expression of IL-1β was obviously down-regulated and the mRNA expression of IL-10 was obviously up-regulated in the EGCG2 and the EGCG3 group (P<0.05 or P<0.01). Compared with the control group, the protein levels of IL-1β and IL-10 in neurons were significantly increased in the OGD/R group (both P<0.01). Compared with the OGD/R group, the protein expression of IL-1β was obviously down-regulated and the protein expression of IL-10 was obviously up-regulated in neurons in the OGD/R+EGCG group (both P<0.01).
Conclusion
EGCG can inhibit the inflammatory reaction induced by cerebral I/R, which may be related to down-regulating the expression of pro-inflammatory factor IL-1β and up-regulating anti-inflammatory factor IL-10.
Keywords: epigallocatechin gallate, ischemia reperfusion, inflammatory reaction, interleukin-1β, interleukin-10
作为茶叶中茶多酚生物活性的主要成分，表没食子儿茶素没食子酸酯(epigallocatechin gallate，EGCG)具有抗氧化、抗辐射、抗肿瘤等作用[1-2]。EGCG可减轻脑缺血导致的大鼠神经功能障碍和组织损伤，缩小脑梗死体积，抑制细胞凋亡[3]；对脑缺血引起的记忆损伤有显著改善作用[4]。笔者前期的研究[5]显示：EGCG能减轻脑缺血再灌注(ischemia/reperfusion，I/R)诱导的鼠脑皮层神经元氧化应激损伤，增强神经元的抗氧化能力。缺血性脑卒中后的炎症反应是神经损伤的重要原因，炎症反应信号通路的激活在I/R的病理生理过程中起关键作用[6]。在脑缺血炎症反应这一复杂的多细胞级联反应中，促炎细胞因子IL-1β与抗炎细胞因子IL-10发挥着重要的调节作用[7-8]。
本研究通过脑I/R模型和氧糖剥夺/再灌注(oxygen-glucose deprivation/reperfusion，OGD/R)细胞模型，观察EGCG对大鼠脑缺血炎症反应和IL-1β、IL-10表达的影响，以期为探索EGCG减轻脑缺血损伤的分子机制提供有价值的线索。
1. 材料与方法
1.1. 材料
EGCG购自美国Sigma公司；髓过氧化物酶(myeloperoxidase，MPO)测试盒购自南京建成生物工程研究所；大鼠IL-1β和IL-10 ELISA试剂盒、鼠抗β-actin单克隆抗体及HRP标记羊抗小鼠IgG二抗为英国Abcam公司产品；引物由生工生物工程上海有限公司合成；反转录试剂盒为美国Thermo Fisher Scientific公司产品；鼠抗IL-1β、IL-10单克隆抗体购自美国Santa Cruz公司。
1.2. 方法
1.2.1. 局灶性脑I/R模型
取体重为280~320 g的雄性SD大鼠，建立大脑中动脉栓塞模型。麻醉大鼠后，作颈部正中切口，钝性分离右颈总动脉、颈内动脉及颈外动脉。从颈外动脉插入尼龙线，沿颈内动脉入颅方向继续前行，造成大脑中动脉栓塞。大鼠缺血1.5 h后将尼龙线拔出，行再灌注24.0 h。
1.2.2. 大脑皮层神经元的分离和培养
按笔者前期的方法[9]进行大脑皮层神经元的分离和培养。取孕18 d左右的SD大鼠胎鼠，在无菌条件下分离大脑皮层，用胰酶在37 ℃下消化，分离所得细胞重悬于含B-27的neurobasal培养基，种植在用多聚-D-赖氨酸预先包被的培养板或培养瓶中。置37 ℃、5% CO2、饱和湿度的细胞培养箱内，体外培养12 d的细胞用于实验。
1.2.3. OGD/R细胞模型
参照笔者前期的方法制备神经元OGD/R模型[9]。
OGD：将细胞培养液换成预先通以95% N2、5% CO2的无糖培养液，而后将神经元置于含95% N2、5% CO2的缺氧密闭环境中，在37 ℃、饱和湿度条件下培养2 h。
R：细胞经OGD损伤 2 h后，将无糖培养液换成葡萄糖浓度为25.0 mmol/L的培养液，重置细胞于37 ℃、5% CO2、饱和湿度培养箱内培养24 h。
1.2.4. 实验分组
实验方案经苏州卫生职业技术学院实验动物伦理委员会批准。SD大鼠随机分成5组：假手术(Sham)组、I/R组、EGCG1组、EGCG2组、EGCG3组。Sham组大鼠除大脑中动脉不予栓塞外，其余处理与I/R组相同。EGCG1组、EGCG2组、EGCG3组大鼠在栓塞前，分别腹腔注射12.5、25.0、50.0 mg/kg的EGCG (EGCG溶于生理盐水)。Sham组大鼠和I/R组大鼠腹腔注射等体积生理盐水。
体外培养的神经元细胞在37 ℃、5% CO2、饱和湿度的培养箱内培养，并随机分成3组：对照(control)组、OGD/R组、OGD/R+EGCG组(OGD/R+50.0 μmol/L EGCG)。Control组除不予OGD处理外，其余同OGD/R组。OGD/R+EGCG组神经元在OGD前用终浓度为50.0 μmol/L的EGCG预处理。
1.2.5. 脑组织含水量的测定
实验结束后，将大鼠断头取脑。取缺血侧大脑称重(湿质量)，然后将其置于110 ℃电干燥箱24 h，再次称重(干质量)。脑组织含水量=(脑湿质量-脑干质量)/脑湿质量×100%。
1.2.6. 脑组织MPO活性的检测
按MPO测试盒说明，用比色法检测缺血区脑组织MPO活性。实验结果以每克脑组织所含酶活力单位表示(U/g)，一个酶活力单位(U)指每分钟分解 1 μmol H2O2。
1.2.7. 脑组织IL-1β和IL-10含量测定
用4 ℃生理盐水在冰浴条件下，制作10%的脑组织匀浆，在4 ℃、4 000 r /min条件下离心10 min，取上清液。用IL-1β、IL-10 ELISA试剂盒测定大鼠脑组织IL-1β、IL-10含量，实验结果以每克脑组织蛋白所含IL-1β、IL-10量表示(ng/g Pro)。
1.2.8. RT-PCR
采用RT-PCR检测IL-1β、IL-10 mRNA表达。提取各组大鼠脑组织总RNA。按试剂盒说明进行反转录：42 ℃ 60 min，70 ℃ 10 min。β-actin正向引物为5'-GTAAAGACCTCTATGCCAACA-3'，反向引物为5'-GGACTCATCGTACTCCTGCT-3'，产物大小为 227 bp。IL-1β正向引物为5'-TGTGATGTTCCCATT-AGAC-3'，反向引物为5'-AATACCACTTGTTGGCT-TA-3'，产物大小为131 bp。IL-10正向引物为5'-CAGTCAGCCAGACCCACAT-3'，反向引物为5'-GGCAACCCAAGTAACCCT-3'，产物大小为141 bp。荧光定量PCR反应：94 ℃ 3 min，94 ℃ 15 s，58 ℃ 30 s，共40个循环。目的基因表达量采用2-ΔΔCt法，根据目的基因和内参基因Ct值计算，结果以与Sham组目的基因表达量的百分比表示。
1.2.9. 蛋白质印迹
采用Western印迹检测IL-1β和IL-10蛋白质的表达水平。提取蛋白质，进行SDS-聚丙烯酰胺凝胶电泳后转移到PVDF膜。于室温下，在含3%牛血清白蛋白的TBST(含0.05% Tween-20 的 TBS）中封闭1 h，分别加入鼠抗IL-1β、IL-10、β-actin单克隆抗体，于4 ℃孵育过夜。用TBST漂洗后，用HRP标记羊抗小鼠IgG二抗在室温下孵育1 h。以β-actin为内参。
1.3. 统计学处理
用SPSS统计软件进行数据分析，实验结果以均数±标准差( ±s)表示，组间比较采用单因素方差分析，以P<0.05为差异有统计学意义。
2. 结 果
2.1. EGCG对I/R大鼠脑组织含水量的影响
与Sham组比较，I/R组、EGCG1组、EGCG2组和EGCG3组大鼠脑组织含水量均明显增加(均P<0.01)。与I/R组比较，EGCG1组、EGCG2组大鼠脑组织含水量差异均无统计学意义(均P>0.05)，EGCG3组大鼠脑含水量明显降低(P<0.01，图1)。
图1.
EGCG对I/R大鼠脑组织含水量的影响
Figure 1 Effect of EGCG on the water content in brain tissues of rats subjected to I/R
**P<0.01 vs the sham group; ††P<0.01 vs the I/R group.
2.2. EGCG对I/R大鼠脑组织MPO活性的影响
与sham组比较，I/R组、EGCG1组、EGCG2组和EGCG3组大鼠脑组织MPO活性均显著增加(均P<0.01)。与I/R组比较，EGCG1组、EGCG2组大鼠脑组织MPO活性无显著变化(均P>0.05)，EGCG3组大鼠脑组织MPO活性显著降低(P<0.01，图2)。
图2.
EGCG对I/R大鼠脑组织MPO活性的影响
Figure 2 Effect of EGCG on the activity of MPO in brain tissues of rats subjected to I/R
**P<0.01 vs the sham group; ††P<0.01 vs the I/R group.
2.3. EGCG对I/R大鼠脑组织IL-1β和IL-10含量的影响
与Sham组比较，I/R组、EGCG1组、EGCG2组和EGCG3组大鼠脑组织IL-1β(均P<0.01)、IL-10含量均显著增加(P<0.05或P<0.01)。与I/R组比较，EGCG1组大鼠脑组织IL-1β、IL-10含量差异均无统计学意义(均P>0.05)；EGCG2组大鼠脑组织IL-1β含量差异无统计学意义(P>0.05)，IL-10含量明显增多 (P<0.01)；EGCG3组大鼠脑组织IL-1β含量显著减少，IL-10含量显著增加(均P<0.01；图3，4)。
图3.
EGCG对I/R大鼠脑组织IL-1β含量的影响
Figure 3 Effect of EGCG on IL-1β content in brain tissues of rats subjected to I/R
**P<0.01 vs the sham group; ††P<0.01 vs the I/R group.
图4.
EGCG对I/R大鼠脑组织IL-10含量的影响
Figure 4 Effect of EGCG on IL-10 content in brain tissues of rats subjected to I/R
*P<0.05,**P<0.01 vs the sham group; ††P<0.01 vs the I/R group.
2.4. EGCG对I/R大鼠脑组织IL-1β和IL-10 mRNA的影响
与Sham组比较，I/R组、EGCG1组、EGCG2组和EGCG3组大鼠缺血区脑组织IL-1β、IL-10 mRNA水平均显著上调(均P<0.01)。与I/R组比较，EGCG1组大鼠缺血区脑组织IL-1β、IL-10 mRNA水平无明显变化(均P>0.05)；EGCG2组、EGCG3组大鼠缺血区脑组织IL-1β mRNA水平显著下调、IL-10 mRNA水平显著上调(P<0.05或P<0.01；图5，6)。
图5.
EGCG对I/R大鼠脑组织IL-1β mRNA的影响
Figure 5 Effect of EGCG on the expression of IL-1β mRNA in cerebral tissues of rats subjected to I/R
**P<0.01 vs the sham group; †P<0.05 vs the I/R group, ††P<0.01 vs the I/R group.
图6.
EGCG对I/R大鼠脑组织IL-10 mRNA的影响
Figure 6 Effect of EGCG on the expression of IL-10 mRNA in cerebral tissues of rats subjected to I/R
**P<0.01 vs the sham group; ††P<0.01 vs the I/R group.
2.5. EGCG对OGD/R大鼠神经元IL-1β和IL-10蛋白质表达的影响
与control组比较，OGD/R组神经元IL-1β及IL-10蛋白质表达水平均显著上调(均P<0.01)。与OGD/R组比较，OGD/R+EGCG组神经元IL-1β蛋白质表达水平明显下调(P<0.01)，IL-10蛋白质表达水平明显上调(P<0.01，图7)。
图7.
EGCG对OGD/R大鼠脑皮层神经元IL-1β和IL-10蛋白质表达的影响
Figure 7 Effect of EGCG on the expression of IL-1β and IL-10 protein in cerebral cortical neurons of rats exposed to OGD/R
**P<0.01 vs the control group; ††P<0.01 vs the OGD/R group.
3. 讨 论
缺血性脑卒中常伴有神经炎症发应，炎症反应可破坏脑内稳态，进一步加重病情[10]。调控神经炎症反应是减轻脑I/R损伤的关键环节，对患者的预后有重要价值。本研究结果显示：I/R引起大鼠脑组织含水量、MPO活性明显增加。50.0 mg/kg EGCG处理能显著降低I/R大鼠脑组织的含水量及MPO活性，提示EGCG可减轻脑I/R导致的神经炎症反应。
IL-1β为IL-1家族成员之一，在炎症反应和免疫应答等过程中扮演重要角色。作为一种多效性的细胞因子，IL-1β是脑缺血损伤介导炎症反应的关键分子之一[7]，能触发多种黏附分子、细胞因子的表达，促进炎症细胞侵入脑实质，进一步加重脑缺血的损害。Diaz-Cañestro等[11]的研究表明：JunD可通过抑制IL-1β减轻I/R导致的小鼠脑损伤。Bai等[12]的研究证实：上调IL-lβ受体拮抗剂的表达能减轻大鼠脑缺血损伤的炎症反应，改善认知功能，发挥神经保护效应。
本研究结果显示：脑缺血1.5 h再灌注24.0 h后，大鼠脑组织IL-1β含量和mRNA水平均升高，提示I/R大鼠神经炎症发应与IL-1β有关。EGCG能下调大鼠脑组织IL-1β mRNA水平，降低IL-1β含量，提示EGCG减轻脑I/R神经炎症反应可能与抑制IL-1β的表达有关。
IL-10是一种重要的抗炎细胞因子[8]，对脑缺血损伤发挥神经保护效应。Liesz等[13]的研究证实：给小鼠脑室注射IL-10能下调脑缺血导致的促炎性级联信号分子表达，缩小小脑梗死灶体积。Fouda等[14]的研究发现：血管紧张素2型受体激动剂C12能减轻脑缺血大鼠的炎症发应、缩小其梗死灶体积、改善其行为和功能，而IL-10抗体可阻断上述效应。
本研究结果显示：大鼠脑组织IL-10 mRNA水平上升，IL-10含量增加，提示脑缺血刺激内源性神经保护因子的产生；EGCG进一步上调了缺血区脑组织IL-10 mRNA水平，增加了IL-10含量，提示EGCG抑制大鼠脑IL-1β的表达可能与其上调IL-10的表达有关。EGCG减轻炎症反应可能与调控缺血性脑损伤内源性保护机制，促进内源性IL-10的表达及抑制IL-1β的表达有关。通过大鼠脑皮层神经元OGD/R模型进一步证实：EGCG能明显下调I/R神经元IL-1β蛋白质水平，上调IL-10蛋白质水平。
综上，IL-10、IL-1β可能是EGCG保护脑缺血损伤的作用靶点。EGCG降低I/R大鼠脑组织含水量及MPO活性，减轻脑缺血损伤炎症反应，可能与其促进抗炎细胞因子IL-10的表达及抑制促炎细胞因子IL-1β的表达有关。
基金资助
江苏省教育厅高校“青蓝工程”([2019]3)；苏州市科技局科技发展计划项目(SYSD2018073)；苏州卫生职业技术学院科技创新团队(SZWZYTD201806)。
This work was supported by the Qing Lan Project in Colleges and Universities of Jiangsu Provincial Education Department ([2019]3), the Science and Technology Development Program of Suzhou Science and Technology Bureau (SYSD2018073), and the Science and Technology Innovation Team of Suzhou Vocational Health College (SZWZYTD201806), China.
利益冲突声明
作者声称无任何利益冲突。
原文网址
http://xbyxb.csu.edu.cn/xbwk/fileup/PDF/2021121325.pdf
参考文献
- 1. Sulistiyani E, Brimson JM, Chansaenroj A, et al. Epigallocatechin-3-gallate protects pro-acinar epithelia against salivary gland radiation injury[J]. Int J Mol Sci, 2021, 22(6): 3162. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 2. Huang YJ, Wang KL, Chen HY, et al. Protective effects of epigallocatechin gallate (EGCG) on endometrial, breast, and ovarian cancers[J]. Biomolecules, 2020, 10(11): 1481. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 3. Park DJ, Kang JB, Koh PO. Epigallocatechin gallate alleviates neuronal cell damage against focal cerebral ischemia in rats[J]. J Vet Med Sci, 2020, 82(5): 639-645. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4. Kaur H, Kumar B, Chakrabarti A, et al. A new therapeutic approach for brain delivery of epigallocatechin gallate: development and characterization studies[J]. Curr Drug Deliv, 2019, 16(1): 59-65. [DOI] [PubMed] [Google Scholar]
- 5. 贺芳, 张轶, 陈尚, 等. EGCG对氧糖剥夺/再灌注神经元氧化应激及Nrf2/HO-1通路的影响[J]. 中南大学学报(医学版), 2018, 43(10): 1041-1047. [DOI] [PubMed] [Google Scholar]; HE Fang, ZHANG Yi, CHEN Shang, et al. Effect of EGCG on oxidative stress and Nrf2/HO-1 pathway in neurons exposed to oxygen-glucose deprivation/reperfusion[J]. Journal of Central South University. Medical Science, 2018, 43(10): 1041-1047. [DOI] [PubMed] [Google Scholar]
- 6. Cao BQ, Tan F, Zhan J, et al. Mechanism underlying treatment of ischemic stroke using acupuncture: transmission and regulation[J]. Neural Regen Res, 2021, 16(5): 944-954. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7. Serhan A, Aerts JL, EWGM Boddeke, et al. Neuroprotection by insulin-like growth factor-1 in rats with ischemic stroke is associated with microglial changes and a reduction in neuroinflammation[J]. Neuroscience, 2020, 426: 101-114. [DOI] [PubMed] [Google Scholar]
- 8. Lyu Q, Pang X, Zhang Z, et al. Microglial V-set and immunoglobulin domain-containing 4 protects against ischemic stroke in mice by suppressing TLR4-regulated inflammatory response[J]. Biochem Biophys Res Commun, 2020, 522(3): 560-567. [DOI] [PubMed] [Google Scholar]
- 9. 贺芳, 叶蓓, 陈建珍, 等. HGF对脑缺血/再灌注大鼠脑iNOS, NO及IL-1β的影响[J]. 中南大学学报(医学版), 2014, 39(1): 23-29. [DOI] [PubMed] [Google Scholar]; HE Fang, YE Bei, CHEN Jianzhen, et al. Influence of hepatocyte growth factor on iNOS, NO and IL-1β in the cerebrum during cerebral ischemia/reperfusion in rats[J]. Journal of Central South University. Medical Science, 2014, 39(1): 23-29. [DOI] [PubMed] [Google Scholar]
- 10. Xu R, Bai Y, Min S, et al. In vivo Monitoring and assessment of exogenous mesenchymal stem cell-derived exosomes in mice with ischemic stroke by molecular imaging[J]. Int J Nanomedicine, 2020, 15: 9011-9023. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 11. Diaz-Cañestro C, Reiner MF, Bonetti NR, et al. AP-1 (activated protein-1) transcription factor JunD regulates ischemia/reperfusion brain damage via IL-1β (interleukin-1β)[J]. Stroke, 2019, 50(2): 469-477. [DOI] [PubMed] [Google Scholar]
- 12. Bai N, Zhang Q, Zhang W, et al. G-protein-coupled estrogen receptor activation upregulates interleukin-1 receptor antagonist in the hippocampus after global cerebral ischemia: implications for neuronal self-defense[J]. J Neuroinflammation, 2020, 17(1): 45. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 13. Liesz A, Bauer A, Hoheisel JD, et al. Intracerebral interleukin-10 injection modulates post-ischemic neuroinflammation: an experimental microarray study[J]. Neurosci Lett, 2014, 579: 18-23. [DOI] [PubMed] [Google Scholar]
- 14. Fouda AY, Pillai B, Dhandapani KM, et al. Role of interleukin-10 in the neuroprotective effect of the Angiotensin Type 2 Receptor agonist, compound 21, after ischemia/reperfusion injury[J]. Eur J Pharmacol, 2017, 799: 128-134. [DOI] [PMC free article] [PubMed] [Google Scholar]