Page 19 - 《中国药房》2022年12期

·药学研究·

蛭龙活血通瘀胶囊对小鼠脑出血及长链非编码RNA和靶基因的

影响 Δ

3 #
1＊
谭睿陟 ，钟 霞 ，王 丽 ，杨思进 ，徐厚平 （1.西南医科大学附属中医医院中西医结合研究中心，四川 泸州
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646000；2.西南医科大学附属中医医院心脑病科，四川 泸州 646000；3.西南医科大学附属中医医院治未病中
心，四川 泸州 646000）
中图分类号 R965；R743.34 文献标志码 A 文章编号 1001-0408（2022）12-1421-09
DOI 10.6039/j.issn.1001-0408.2022.12.03

摘 要 目的 研究蛭龙活血通瘀胶囊对小鼠脑出血及长链非编码RNA（LncRNA）和靶基因的影响。方法 将24只C57BL/6雄性
小鼠随机均分为假手术1组、模型1组和蛭龙活血通瘀胶囊低、高剂量组（0.35、1.40 g/kg），尾状核注射胶原酶以建立脑出血模型。
术后1 h，各药物组小鼠灌胃相应药液，假手术1组和模型1组小鼠灌胃生理盐水，每天1次，连续3 d。通过苏木精-伊红和尼氏染
色观察各组小鼠脑组织的形态学变化，通过免疫组化法、Western blot法和实时定量聚合酶链反应（PCR）法检测其脑组织中白细胞
介素1β（IL-1β）和肿瘤坏死因子α（TNF-α）蛋白及mRNA的表达水平。另将9只C57BL/6雄性小鼠随机均分为假手术2组、模型2
组和干预组（蛭龙活血通瘀胶囊1.40 g/kg），按上述方法建模、给药，分离全脑组织，提取总RNA后测序并分析差异LncRNA，进行
基因本体（GO）富集，预测有效LncRNA和靶基因，并通过实时定量PCR法进行验证。结果 与模型1组比较，蛭龙活血通瘀胶囊
低、高剂量组小鼠脑组织病理损伤明显改善，脑组织中IL-1β、TNF-α蛋白及mRNA的表达均显著下调（P＜0.05）。筛选出2个有效
LncRNA 体内验证结果（LncRNA-Dlst-211 在模型 2 组中高表达，经蛭龙活血通瘀胶囊干预后显著下调；LncRNA-Dlst-211 靶
基因 Rps6kl1 和 LncRNA-MSTRG.8169.4 在模型 2 组中低表达，经蛭龙活血通瘀胶囊干预后显著上调），与测序结果一致。结
论 蛭龙活血通瘀胶囊能改善脑出血模型小鼠的脑损伤及炎症反应，其作用机制可能与下调 LncRNA-Dlst-211 表达，上
调 LncRNA-MSTRG.8169.4、Rps6kl1表达有关。
关键词 蛭龙活血通瘀胶囊；脑出血；炎症；长链非编码RNA；转录组测序

Effects of Zhilong huoxue tongyu capsules on intracerebral hemorrhage，LncRNA and target genes in mice
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TAN Ruizhi ，ZHONG Xia ，WANG Li ，YANG Sijin ，XU Houping（1. Research Center of Traditional Chinese
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Medicine and Western Medicine Integration，the Affiliated Traditional Chinese Medicine Hospital of Southwest
Medical University，Sichuan Luzhou 646000，China；2. Dept. of Cardiovascular and Cerebrovascular Disease，
the Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University，Sichuan Luzhou 646000，
China；3. Preventive Treatment Center，the Affiliated Traditional Chinese Medicine Hospital of Southwest
Medical University，Sichuan Luzhou 646000，China）
ABSTRACT OBJECTVE To study the effects of Zhilong huoxue tongyu capsules on intracerebral hemorrhage，long non-coding
RNA（LncRNA）and its target genes in mice. METHODS Twenty-four male C57BL/6 mice were randomly divided into sham
operation group 1，model group 1 and Zhilong huoxue tongyu capsule low-dose and high-dose groups （0.35，1.40 g/kg）.
Collagenase was injected into the caudate nucleus to construct the model of intracerebral hemorrhage. One hour after the operation，
the mice in each treatment group were given the corresponding medicinal solution，and the mice in the sham operation group 1 and
the model group 1 were given normal saline intragastrically，once a day，for 3 consecutive days. The morphological changes of the
brain tissue of the mice in each group were observed by hematoxylin-eosin staining and Nissl staining. The protein and mRNA
expression of interleukin-1β（IL-1β）and tumor necrosis factor-α（TNF-α）in the brain tissue were detected by immunohistoche-
mistry，Western blot and real-time quantitative polymerase chain reaction （PCR）. In addition，9 of male C57BL/6 mice were
randomly divided into sham operation group 2，model group 2
Δ 基金项目：国家自然科学基金青年科学基金项目（No.81703905）；
and intervention group（Zhilong huoxue tongyu capsule 1.40
国家中医临床基地项目（No.国中医药科技函〔2018〕131号）；四川省省
g/kg）. The mice were modeled and administered according to
级科技计划项目（No.2019YFS0543）
＊高级实验师。研究方向：中西医结合防治炎症和纤维化。 the above method，and then the whole brain tissue of mice in
E-mail：tanruizhi627@swmu.edu.cn each group was isolated， total RNA was extracted and
# 通信作者：主任中医师，博士。研究方向：中西医结合防治心脑 sequenced，followed by analyzing the different LncRNA. Gene
血管疾病。E-mail：xuhpswmu@163.com ontology（GO）enrichment was performed to predict effective

中国药房 2022年第33卷第12期 China Pharmacy 2022 Vol. 33 No. 12 ·1421 ·