文章摘要
木艳玲, ，刘扬, ，李媛, ，薛孟周,.丹酚酸B对小鼠脑出血的神经保护作用研究[J].神经损伤功能重建,2023,(8):435-440
丹酚酸B对小鼠脑出血的神经保护作用研究
Neuroprotective Effect of Salvianolic Acid B on Intracerebral Hemorrhage in Mice
  
DOI：
中文关键词: 脑出血  丹酚酸B  炎症  神经保护  血脑屏障
英文关键词: intracerebral hemorrhage  Salvianolic acid B  inflammatory  neuroprotection  blood-brain barrier
基金项目:国家自然科学基金 项目（No. 8207052 870）； 国家自然科学基金 重点国际（地区）合 作研究项目（No. 8 1520108011）； 国家重点研发计划 项目（No. 2018YF C1312200）
作者单位
木艳玲1,2 ，刘扬1,2 ，李媛1,2 ，薛孟周1,2 1. 郑州大学第二附 属 医 院 脑 血 管 病 科 2. 郑州大学医学科 学院 
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中文摘要:
      目的：探讨丹酚酸B（SalB）对小鼠脑出血（ICH）的神经保护作用及机制。方法：C57BL/6J雄性小鼠 108只随机分为Sham组，ICH+Vehicle组，ICH+SalB组，每组36只。Ⅶ型胶原酶制备ICH模型，ICH+SalB 组术后2 h尾静脉注射SalB 30 mg/kg，Sham组和ICH+Vehicle组尾静脉注射等量生理盐水，连续3 d。术后 第3天，采用mNSS和转角实验观察行为学指标；HE染色观察脑组织损伤情况；脑含水量检测评估脑组织 水肿；伊文思蓝（EB）外渗试验评估血脑屏障渗透性；Western blot检测ZO-1和Occludin蛋白表达水平评估 血脑屏障完整性，检测MMP-9和IL-1β蛋白表达水平评估炎症因子水平；TUNEL染色评估凋亡细胞数量； Iba1和MPO免疫荧光染色评估小胶质细胞活化和中性粒细胞渗出情况。结果：mNSS评分结果显示ICH+ SalB组神经功能缺损低于ICH+Vehicle组（P＜0.05）；转角实验结果显示ICH+SalB组右转次数低于ICH+ Vehicle 组（P＜0.05）；HE 染色结果显示 ICH+SalB 组脑组织损伤、血肿面积较 ICH+Vehicle 组降低（P＜ 0.05）；脑含水量结果显示ICH+SalB组脑水肿程度轻于ICH+Vehicle组（P＜0.05）；EB外渗结果显示ICH+ SalB组EB外渗低于ICH+Vehicle组（P＜0.05）；Western blot结果显示ICH+SalB组MMP-9、IL-1β表达低于 ICH+Vehicle组（P＜0.05），ICH+SalB组ZO-1、occludin表达高于ICH+Vehicle组（P＜0.05）；TUNEL结果显 示ICH+SalB组凋亡细胞数量低于ICH+Vehicle组（P＜0.05）；免疫荧光检测显示ICH+SalB组Iba1和MPO 水平均低于ICH+Vehicle组（P＜0.05）。结论：SalB可能通过减少小鼠ICH后的脑水肿、降低炎症因子、稳定 血脑屏障的机制改善神经功能，发挥神经保护作用。
英文摘要:
      This aim of this study was to investigate the neuroprotective effect of salvianolic acid B (SalB) on intracerebral hemorrhage (ICH) in mice, and to elucidate the underlying mechanism. Methods: A total of 108 C57BL/6J male mice were randomly divided into the Sham, ICH+Vehicle, and ICH+ SalB groups, with 36 mice per group. The ICH model was prepared with type Ⅶ collagenase. Mice in the ICH+SalB group were injected with 30 mg/kg SalB via the tail vein at 2 hours after operation, and mice in the Sham and ICH+Vehicle groups were similarly injected with the same amount of normal saline via the tail vein for 3 consecutive days. On the 3rd day after operation, the modified neurological severity score (mNSS) and corner turn test were conducted to assess behavioral indexes. Further, hematoxylin and eosin (HE) staining was used to observe injury to the brain tissues, brain water content was measured to evaluate cerebral edema, and Evens blue (EB) extravasation test was used to evaluate the permeability of the blood-brain barrier (BBB). In addition, western blot was performed to detect the expression levels of ZO-1 and Occludin to evaluate the integrity of the BBB and to detect the expression levels of MMP-9 and IL-1β as exemplar inflammatory factors. Finally, TUNEL staining was performed to evaluate the number of apoptotic cells, and immunofluorescence staining using Iba1 and MPO was adopted to evaluate microglia activation and neutrophil exudation. Results: The results of mNSS testing showed that the neurologic deficit degree in the ICH+SalB group was lower than that in the ICH+Vehicle group (P<0.05), while in the corner turn test, the number of right turns in the ICH+SalB group was less than that in ICH+Vehicle group (P<0.05). HE staining showed that the degree of brain tissue injury and hematoma area in the ICH+SalB group were reduced compared to the ICH+Vehicle group (P<0.05). Brain water content analysis showed that the degree of cerebral edema in the ICH+SalB group was less than that in the ICH+Vehicle group (P<0.05), and the EB extravasation level in the ICH+SalB group was lower than that in ICH+Vehicle group (P< 0.05). Further, the western blot results showed that the expression levels of MMP-9 and IL-1β in ICH+SalB group were lower than those in the ICH+Vehicle group (P<0.05), while the expression levels of ZO-1 and Occludin in ICH+SalB group were higher than those in the ICH+Vehicle group (P<0.05). TUNEL staining showed that the number of apoptotic cells in the ICH+SalB group was lower than that in ICH+Vehicle group (P<0.05). Finally, the immunofluorescence staining results showed that the Iba1 and MPO levels were lower in the ICH+SalB group than in the ICH+Vehicle group (P<0.05). Conclusion: Overall, our results indicate that SalB may improve neurological function and play a neuroprotective role by reducing cerebral edema, the level of inflammatory factors, and stabilizing the blood-brain barrier in mice with ICH.
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