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ABSTRACT OBJECTIVE:To study preventive effect and mechanism of ginsenoside Rg1 on focal cerebral ischemia-reperfusion
injury (CIRI) model rats. METHODS:Totally 78 SD rats were randomly divided into sham operation group,model group,
butylphthalide control group(positive control,10 mL/kg),ginsenoside Rg1 low-dose,medium-dose and high-dose groups(10,
20,40 mg/kg),with 13 rats in each group. Administration groups were give relevant medicine intraperitoneally,sham operation
group and model group were given constant volume of normal saline intraperitoneally,once a day,for consecutive 7 d. After
medication,except for the sham operation group,focal CIRI model was induced by middle cerebral artery occlusion(MCAO)
method in other groups. After modeling,neurological deficit scoring was performed according to the modified neurological difict
scoring standard; TTC staining was used to detected the percentage of cerebral infarction of rats;the cerebral water content was
measured by dry/wet weight method;serum contents of IL-1β and IL-6 were detected by ELISA;the protein expressions of p-p38
MAPK and p-NF-κ B p65 in cerebral tissue were determined by immunohistochemistry and Western blotting assay. RESULTS:
Compared with sham operation,neurological deficits score,percentage of cerebral infarction and cerebral water content,serum
contents of IL-1β and IL-6,positive expression numbers of cells and protein expressions of p-p38 MAPK and p-NF-κB p65 in
cerebral tissue were increased significantly in model group(P<0.05 or P<0.01). Compared with model group,above index levels
of administration groups were all decreased significantly (P<0.05 or P<0.01),and the effect of ginsenoside Rg1 had a
dose-dependent trend;there was no significant difference of all above indexes between ginsenoside Rg1 middle-dose,high-dose
groups and butylphthalide control group (P>0.05). CONCLUSIONS:Ginsenoside Rg1 has a certain preventive effect on focal
CIRI model rats,the mechanism of which may be associated with down-regulating the protein expression of p-p38 MAPK and
p-NF-κB p65,inhibiting the release of inflammatory factors such as IL-1β and IL-6.
KEYWORDS Ginsenoside Rg1;p-p38 MAPK;p-NF-κB p65;Cerebral ischemia-reperfusion injury;Inflammation;Rats
缺血性脑卒中具有高复发率、高致残率和高致死率 觉科技股份有限公司);Primo Start 型荧光显微镜(德国
的特点,严重危害人类健康。目前,对于该病的治疗原 Zeiss 公司);Matrx VIP 3000 型动物麻醉机(美国 Mid-
则主要为溶解血栓、恢复血液供应,但血液再通时经常 mark公司)。
会导致脑缺血再灌注损伤(Cerebral ischemia reperfusion 1.2 药品与试剂
[1]
injury,CIRI) 。减轻再灌注损伤已成为缺血性脑血管 人参皂苷Rg1对照品(吉林大学有机化学教研室,批
疾病治疗的重要环节。研究显示,炎症是CIRI的主要原 号:201810,纯度:>95%);丁苯酞注射液(石药集团恩
因之一 [2-3] 。从抑制炎症反应方面探讨药物是否对 CIRI 必普制药有限公司,批号:618180813,规格:每100 mL中
有防治作用已成为目前研究的热点。人参皂苷Rg1为传 含丁苯酞 25 mg、氯化钠 0.9 g);兔抗磷酸化 p38 MAPK
统中药人参的主要活性成分之一,以往研究证实,人参 (p-p38 MAPK)多克隆抗体、兔抗磷酸化 NF-κB p65
皂苷Rg1能够改善缺血再灌注损伤症状,发挥神经保护作 (p-NF-κB p65)多克隆抗体、兔抗磷酸甘油醛脱氢酶
用,其作用机制可能与抗细胞凋亡、抗氧化作用有关 [4-5] 。 (GAPDH)多克隆抗体(英国 Abcam 公司);异硫氰酸荧
目前,也有文献证实,人参皂苷Rg1可通过抑制炎症反应 光素(FITC)标记的免疫球蛋白 G(IgG)二抗(北京中杉
发挥对中脑黑质多马胺能神经元的保护作用 [6-7] 。但是 金桥生物技术有限公司);白细胞介素1β(IL-1β)、IL-6酶
关于人参皂苷 Rg1对 CIRI 中的炎症反应是否有抑制作 联免疫吸附(ELISA)检测试剂盒(美国R&D公司,批号:
用,目前还不甚清楚。p38 丝裂原活化蛋白激酶(p38 均为201901);二喹啉甲酸(BCA)蛋白浓度测定试剂盒(北
MAPK)/核因子-κB(NF-κB)是目前较为经典的炎症通路, 京鼎国昌盛生物技术有限责任公司,批号:93H001100);
[8]
可以调控多种炎症因子的表达,从而调节炎症反应 。 增强化学发光显影剂(ECL,上海碧云天生物技术有限公
鉴于此,本研究拟通过研究人参皂苷 Rg1对局灶性 CIRI 司);2,3,5-氯化三苯基四氮唑(TTC)染色剂(美国 Sig-
模型大鼠炎症反应的影响,并通过p38 MAPK/NF-κB通 ma公司);4′,6-二脒基-2-苯基吲哚(DAPI)染色剂(美国
路探索其作用机制,为治疗缺血性脑卒中疾病的新药开 Genview公司);其余试剂均为分析纯,水为超纯水。
发提供实验基础。 1.3 动物
1 材料 健康清洁级 SD 大鼠 78 只,雄性,体质量(280±50)
1.1 仪器 g,由锦州医科大学实验动物中心提供,实验动物使用许
DNM-9602G 型酶标分析仪(广州沪瑞明仪器公 可证号:SYXK(辽)2017-0003。饲养环境温度为 20~
司);PowerPac 3000 型电泳仪、Trans-BlotSDcell 型半干 25 ℃、相对湿度为 50%~60%,饮用水为无菌纯净水。
转印仪(美国Bio-Rad公司);Vibra Cell Vcx105型超声波 本研究已通过锦州医科大学实验动物伦理委员会审核
细胞粉碎机(美国 Sonics 公司);CM1900 型冰冻切片机 (批件编号:2018014)。实验过程中对动物的处置符合
(德国 Leica 公司);YZ20P6 型手术显微镜(苏州六六视 科学技术部颁发的《关于善待实验动物的指导性意见》
·1288 · China Pharmacy 2020 Vol. 31 No. 11 中国药房 2020年第31卷第11期