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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
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| 篇名： | 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 |
| TITLE： | Effects of Zhilong huoxue tongyu capsules on intracerebral hemorrhage ，LncRNA and target genes in mice |
| 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、1.40g/kg），尾状核注射胶原酶以建立脑出血模型。术后1h，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3d。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、Westernblot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1.40g/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。 |
| 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 C 57BL/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 C 57BL/6 mice were randomly divided into sham operation group 2，model group 2 and intervention group （Zhilong huoxue tongyu capsule 1.40 g/kg）. The mice were modeled and administered according to 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 sequenced，followed by analyzing the different LncRNA. Gene ontology（GO）enrichment was performed to predict effective LncRNA and target genes ，and verified by real-time quantitative PCR. RESULTS Compared with model group 1，the brain tissue pathological damages were significantly improved in Zhilong huoxue tongyu capsule low-dose a nd high-dose groups ，and the IL -1β， TNF-α protein and mRNA expression in brain tissue were significantly decreased （P＜0.05）. Two effective LncRNAs were screened out. The results of in vivo verification test （LncRNA-Dlst-211 was highly expressed in model group 2，and significantly down-regulated after the intervention of Zhilong huoxue tongyu capsules ； LncRNA-Dlst-211 target genes Rps 6kl1 and LncRNA-MSTRG.8169.4 were expressed weakly in model group 2，and strongly up-regulated after intervention ）were consistent with the sequencing results. CONCLUSIONS Zhilong huoxue tongyu capsules can improve the brain injury and inflammatory response in intracerebral hemorrhage model mice ，and its mechanism may be related to down-regulating the expression of LncRNA-Dlst-211 and up-regulating the expression of LncRNA-MSTRG. 8169.4 and Rps 6kl1. |
| 期刊： | 2022年第33卷第12期 |
| 作者： | 谭睿陟，钟霞，王丽，杨思进，徐厚平 |
| AUTHORS： | TAN Ruizhi ，ZHONG Xia，WANG Li，YANG Sijin，XU Houping |
| 关键字： | 蛭龙活血通瘀胶囊；脑出血；炎症；长链非编码RNA；转录组测序 |
| KEYWORDS： | Zhilong huoxue tongyu capsules ；intracerebral hemorrhage ；inflammation；long non-coding RNA ；transcriptome |
| 阅读数： | 363 次 |
| 本月下载数： | 5 次 |
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