Department of Neurosurgery, First Affiliated Hospital, Soochow University, 215000 Suzhou, Jiangsu, China.
Department of Neurosurgery, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, 200233 Shanghai, China.
Discov Med. 2023 Aug;35(177):565-575. doi: 10.24976/Discov.Med.202335177.57.
Cerebral ischemia-reperfusion (CI/R) injury is induction of blood flow restoration after an ischemic stroke. Gentiopicroside (GPC) is the principal active secoiridoid glycoside of . This research aimed to illuminate the function of GPC and its mechanism in CI/R injury.
After CI/R injury models were constructed, GPC (25, 50 or 100 mg/kg) was then administered by gavage to rats. Rats were grouped into Sham, CI/R, CI/R+25 mg/kg GPC, CI/R+50 mg/kg GPC, and CI/R+100 mg/kg GPC. Neuronal cells were exposed to oxygen-glucose deprivation and reperfusion (OGD/R) injury to establish ischemic-like conditions , and cells were further treated with 25, 50, or 100 μM GPC. Cells were grouped into control, OGD/R, OGD/R+25 μM GPC, OGD/R+50 μM GPC, and OGD/R+100 μM GPC. GPC's function on rat cerebral injury, angiogenesis, oxidative stress, neuronal injury and immune dysfunction was estimated using hematoxylin-eosin staining, Western blot, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining, commercial kits and enzyme linked-immunosorbent assay. Meanwhile, GPC's mechanism in CI/R injury was examined via Western blot. GPC's function was estimated via Cell Counting Kit-8 assay, 5-ethynyl-2'-deoxyuridine (EdU) staining, flow cytometry.
GPC alleviated cerebral injury through decreasing cerebral infarction volume, cerebral indexes, brain water contents ( < 0.05). GPC reduced oxidative stress and boosted cerebral angiogenesis in CI/R rats ( < 0.05). Meanwhile, GPC weakened neuronal cell apoptosis, and decreased neuron-specific enolase and S100beta protein levels in CI/R rats. GPC reduced inflammatory cytokines contents in serum and brain tissues of CI/R rats ( < 0.05). Moreover, GPC increased the viability and proliferation in OGD/R-treated neuronal cells, but decreased cell apoptosis ( < 0.05). Mechanistically, GPC upregulated vascular endothelial growth factor (VEGF) and phosphorylated nuclear factor E2-related factor 2 (p-Nrf2) levels in CI/R rat brain tissues ( < 0.05).
GPC reduced cerebrovascular angiogenesis, neuronal injury and immune disorder in CI/R injury through elevating VEGF and p-Nrf2.
脑缺血再灌注（CI/R）损伤是缺血性脑卒中后血流恢复引起的。龙胆苦苷（GPC）是獐牙菜苦苷的主要活性环烯醚萜类糖苷。本研究旨在阐明 GPC 在 CI/R 损伤中的作用及其机制。
构建 CI/R 损伤模型后，通过灌胃给予大鼠 GPC（25、50 或 100mg/kg）。大鼠分为假手术组、CI/R 组、CI/R+25mg/kg GPC 组、CI/R+50mg/kg GPC 组和 CI/R+100mg/kg GPC 组。将神经元细胞暴露于氧葡萄糖剥夺再灌注（OGD/R）损伤中，建立类似缺血的条件，并进一步用 25、50 或 100μMGPC 处理细胞。细胞分为对照组、OGD/R 组、OGD/R+25μMGPC 组、OGD/R+50μMGPC 组和 OGD/R+100μMGPC 组。通过苏木精-伊红染色、Western blot、末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记（TUNEL）染色、商业试剂盒和酶联免疫吸附试验评估 GPC 对大鼠脑损伤、血管生成、氧化应激、神经元损伤和免疫功能障碍的作用。同时，通过 Western blot 检测 GPC 在 CI/R 损伤中的作用机制。通过细胞计数试剂盒-8 测定法、5-乙炔基-2'-脱氧尿苷（EdU）染色和流式细胞术评估 GPC 的功能。
GPC 通过降低脑梗死体积、脑指数和脑含水量来减轻脑损伤（<0.05）。GPC 降低了 CI/R 大鼠的氧化应激并促进了脑内血管生成（<0.05）。同时，GPC 减弱了 CI/R 大鼠的神经元细胞凋亡，并降低了神经元特异性烯醇化酶和 S100β 蛋白水平。GPC 降低了 CI/R 大鼠血清和脑组织中炎症细胞因子的含量（<0.05）。此外，GPC 增加了 OGD/R 处理神经元细胞的活力和增殖，但减少了细胞凋亡（<0.05）。机制上，GPC 上调了 CI/R 大鼠脑组织中血管内皮生长因子（VEGF）和磷酸化核因子 E2 相关因子 2（p-Nrf2）水平（<0.05）。
GPC 通过提高 VEGF 和 p-Nrf2 水平，减少了 CI/R 损伤中的脑血管生成、神经元损伤和免疫紊乱。
