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基于 Notch 信号通路的黄芩素抑制急性肺栓塞模型大鼠血小板

聚集及肺组织保护作用机制研究 Δ

＊
#
吴忠勇 ，王金忠 ，周 森，林 明，王小智（海南医学院第二附属医院重症医学科，海口 570311）
中图分类号 R285 文献标志码 A 文章编号 1001-0408（2020）09-1091-07
DOI 10.6039/j.issn.1001-0408.2020.09.13

摘 要 目的：探索黄芩素抑制急性肺栓塞模型大鼠血小板聚集及肺组织保护作用的机制。方法：将36只大鼠随机分为正常对
照组（n＝6）和造模组（n＝30），造模组大鼠采用自体血栓法复制急性肺栓塞模型，正常对照组大鼠行假手术。将造模成功的30只
大鼠随机分为模型对照组、阳性药物组（低分子肝素钙0.01 mL/kg，皮下注射）和黄芩素低、中、高剂量组（25、50、100 mg/kg，腹腔注
射），每组6只。正常对照组和模型对照组大鼠腹腔注射等体积生理盐水，各给药组大鼠给予相应药物，每天1次，连续给药7 d。
给药结束后，检测大鼠血浆二磷酸腺苷（ADP）和花生四烯酸（AA）活化后的血小板聚集率及血小板活化指数（RPI）；采用苏木精-
伊红染色法观察大鼠肺组织病理学变化；采用酶联免疫吸附法检测大鼠血清中血小板活化标志物血小板颗粒膜蛋白（CD62P）和
溶酶体颗粒膜蛋白（CD63）以及生长分化因子15（GDF-15）、N端B型利钠肽（NT-proBNP）水平；采用实时荧光定量聚合酶链式反
应法检测大鼠肺组织中Notch2、Notch3及Notch信号配体DLL1、JAG2 mRNA表达水平；分别采用免疫组化法和Western blotting
法检测大鼠肺组织中Notch2、Notch3、DLL1、JAG2蛋白表达水平。结果：与正常对照组比较，模型对照组大鼠血浆ADP活化后血
小板聚集率、AA活化后血小板聚集率、RPI值以及血清中CD62P、CD63、GDF-15、NT-proBNP水平均显著升高（P＜0.05）；肺组织
处于严重炎性浸润状态；肺组织中Notch2、Notch3、DLL1、JAG2 mRNA及蛋白表达水平均显著降低（P＜0.05）。与模型对照组比
较，黄芩素各剂量组大鼠上述指标变化均显著改善（P＜0.05）。结论：黄芩素能够降低急性肺栓塞模型大鼠血小板聚集，改善大鼠
肺组织病理状态；其机制可能与激活Notch信号通路有关。
关键词 黄芩素；急性肺栓塞；Notch信号通路；血小板聚集；大鼠；机制

Study on Mechanism of Platelet Aggregation Inhibitory Effects and Lung Tissue Protective Effects of
Baicalein in Model Rats with Acute Pulmonary Embolism Based on Notch Signaling Pathway
WU Zhongyong，WANG Jinzhong，ZHOU Sen，LIN Ming，WANG Xiaozhi（Dept. of Critical Medicine，the
Second Affiliated Hospital of Hainan Medical College，Haikou 570311，China）

ABSTRACT OBJECTIVE：To explore the mechanism of baicalein platelet aggregation inhibitiory effect and lung tissue
protective effect of baicalein in model rats with acute pulmonary embolism. METHODS：Totally 36 rats were randomly divided into
normal control group（n＝6）and modeling group（n＝30）. The acute pulmonary embolism model was established by autologous
thrombus replication in modeling group，and the sham operation of rats in normal control group was carried out. After modeling，
30 model rats were randomly divided into model control group，positive drug group（low molecular weight heparin calcium 0.01
mL/kg，subcutaneous injection），baicalein low-dose，middle-dose and high-dose groups （25，50，100 mg/kg，intraperitoneal
injection），with 6 rats in each group. Normal control group and model control group were intraperitoneally injected constant
volume of normal saline；administration groups were given relevant medicine，once a day，for consecutive 7 d. After medication，
platelet aggregation rates of rats after activated with adenosine diphosphate （ADP） and arachidonic acid （AA） and platelet
activation index（RPI）were detected；lung histopathology was observed by HE staining；serum platelet activation markers granule
membrane（CD62P）and lysosomal membrane glycoprotein（CD63），growth differentiation factor-15（GDF-15）and N-terminal
B-type natriuretic peptide（NT-proBNP）were measured by ELISA. The mRNA expression levels of Notch2，Notch3 and Notch
signaling ligand PLL1，JAG2 were detected by RT-PCR method. The protein expression levels of Notch2，Notch3，DLL1 and
JAG2 in lung tissue were detected by immunohistochemistry and Western blotting assay. RESULTS：Compared with normal control
group，plasma ADP-activated platelet aggregation rate，AA-activated platelet aggregation rate，RPI，serum levels of CD62P，
CD63，GDF-15 and NT-proBNP were increased significantly （P＜0.05）. The lung tissue of rats was in a state of severe
inflammatory infiltration. mRNA and protein expression levels of Notch2，Notch3，DLL1 and JAG2 in lung tissue decreased
significantly （P＜0.05）. Compared with model control group，changes of above indexes of rats were improved significantly in
baicalein groups（P＜0.05）. CONCLUSIONS：Baicalein can reduce platelet aggregation and improve the pathological state of lung
tissue in rats with acute pulmonary embolism. Its mechanism
Δ 基金项目：海南省自然科学基金面上项目（No.SQ2018MSXM-
may be related to activating Notch signal pathway.
0270）
KEYWORDS Baicalein； Acute pulmonary embolism；
＊主治医师。研究方向：脓毒血症。E-mail：18610349@qq.com
# 通信作者：主治医师，硕士。研究方向：脓毒血症。E-mail：wj- Notch signal pathway；Platelet aggregation；Rat；Mechanism
zh269@163.com

中国药房 2020年第31卷第9期 China Pharmacy 2020 Vol. 31 No. 9 ·1091 ·