Breviscapine Regulates the mRNA and Protein Levels of CINC-1 and CINC-2 in the Right Ventricle of Rats with Acute Pulmonary Embolism
-
摘要:
目的 观察灯盏花素对大鼠肺栓塞模型右心室CINC-1、CINC-2的mRNA及蛋白水平有无影响。 方法 复制大鼠肺栓塞模型，灯盏花素低、高剂量组及低分子肝素组给予腹腔注射相应药物，取肺组织做肺栓塞病理检测，右心室组织做Q-PCR及Western blot测定各组CINC-1、CINC-2的基因及蛋白水平。 结果 与假手术组比较，模型组CINC-1、CINC-2的基因表达明显升高（P < 0.01），灯盏花素低、高剂量组及低分子肝素组可以显著下调CINC-1、CINC-2的基因水平（P < 0.01），灯盏花素高剂量组在下调CINC-1基因书水平上优于低分子肝素组（P < 0.05）；与假手术组比较，模型组CINC-1、CINC-2的蛋白表达明显升高（P < 0.01），灯盏花素低、高剂量组可以明显下调CINC-1的蛋白水平（P < 0.01），灯盏花素高剂量组在下调CINC-1蛋白表达水平上优于低分子肝素组（P < 0.05）；灯盏花素高剂量组可以明显下调CINC-2的蛋白水平（P < 0.01）。 结论 灯盏花素可以下调急性肺栓塞大鼠右心室 CINC-1、CINC-2 的基因和蛋白表达水平， 可能发挥抗肺栓塞炎症的作用。 Abstract:Objective To observe whether breviscapine has any effect on the mRNA and protein levels of CINC-1 and CINC-2 in the right ventricle of rats with pulmonary embolism. Methods The rat model of pulmonary embolism was replicated, and rats in the breviscapine low-dose, high-dose and low-molecular-weight heparin groups were given intraperitoneal injection of corresponding drugs, lung tissue samples were collected for pathological detection of pulmonary embolism, and right ventricular tissue samples were determined by Q-PCR and Western blot to determine CINC-1, CINC-2 gene and protein levels in each group. Results Compared with the sham operation group, the gene expression of CINC-1 and CINC-2 in the model group was significantly upregulated （P < 0.01）. The low-dose, high-dose and low-molecular-weight heparin groups of breviscapine could significantly down-regulate the gene expression of CINC-1 and CINC-2 （P < 0.01）. The high-dose breviscapine group was better than the low-molecular-weight heparin group in down-regulating CINC-1 gene level （P < 0.05）. Compared with the sham operation group, the protein expressions of CINC-1 and CINC-2 in the model group were significantly increased （P < 0.01）. The low-dose and high-dose groups of breviscapine could significantly down-regulate the protein level of CINC-1 （P < 0.01）, and the high-dose group of breviscapine was better than the low-molecular-weight heparin group in down-regulating the protein expression of CINC-1 （P < 0.05）. And the high dose of breviscapine can significantly down-regulate the protein level of CINC-2 （P < 0.01）. Conclusion Breviscapine may play an anti-inflammatory role in pulmonary embolism by down-regulating the gene and protein expression of CINC-1 and CINC-2 in the right ventricle of rats with acute pulmonary embolism. -
Key words:
- Breviscapine /
- Pulmonary embolism /
- CINC-1 /
- CINC-2
-
图 1 给药后大鼠心率、呼吸变化
与假手术组比较，*P < 0.05， **P < 0.01; 与模型组比较，#P < 0.05， ##P < 0.01。采用 Graphpad Prism 8.0 作柱状图，数据用均数±标准差表示，n = 6。 假手术组（Control），模型组（Model），灯盏花素低剂量组（Bre10 g/L），灯盏花素高剂量组（Bre20 g/L），低分子肝素阳性药组（LMWH 10.250 IU/L）。
Figure 1. Changes in heart rate and respiration in rats after administration
图 2 各组大鼠肺组织病理检查
a：假手术组（Control）；b：模型组（Model）；ｃ：灯盏花素低剂量组（Bre10 g/L）；ｄ：灯盏花素高剂量组（Bre20 g/L）；ｅ：低分子肝素阳性药组（LMWH 10.250 IU/L）；HE染色。
Figure 2. Pathological examination of lung tissue of rats in each group
图 3 Q-PCR检测灯盏花素干预肺栓塞大鼠右心室CINC-1、CINC-2基因表达
与假手术组比较，**P < 0.01；与模型组比较，##P < 0.01；与低分子肝素组比较，&P < 0.05。采用Graphpad Prism 8.0作柱状图，数据用均数±标准差表示，n = 6。 假手术组（Control），模型组（Model），灯盏花素低剂量组（Bre10 mg/mL），灯盏花素高剂量组（Bre20 mg/mL），低分子肝素阳性药组（LMWH 10.250 IU/L）。
Figure 3. Q-PCR detection of right ventricular CINC-1 and CINC-2 gene expression in rats with breviscapine intervention with pulmonary embolism
图 4 WB检测肺栓塞大鼠右心室CINC-1、CINC-2蛋白表达
Figure 4. The protein expression of CINC-1 and CINC-2 in the right ventricle of rats with pulmonary embolism detected by WB
图 5 WB检测灯盏花素干预肺栓塞大鼠右心室CINC-1、CINC-2蛋白表达
与假手术组比较，**P < 0.01；与模型组比较，##P < 0.01；与低分子肝素组比较，&P < 0.05。采用Graphpad Prism 8.0作柱状图，数据用均数±标准差表示，n = 6。 假手术组（Control），模型组（Model），灯盏花素低剂量组（Bre10 g/L），灯盏花素高剂量组（Bre20 g/L），低分子肝素阳性药组（LMWH 10.250 IU/L）。
Figure 5. WB detection of right ventricular CINC-1 and CINC-2 protein expression in rats with breviscapine intervention with pulmonary embolism
表 1 灯盏花素对大鼠肺栓塞影响的量效关系（
$\bar x \pm s $ ）Table 1. Dose-effect relationship of breviscapine on pulmonary embolism in rats （
$\bar x \pm s $ ）组别 假手术组 模型组 灯盏花素组 大鼠（只） 5 5 5 5 5 灯盏花素剂量（g/L） 0 0 10 20 50 心率（次/min） 358 ± 9 418 ± 27** 390.5 ± 9# 366 ± 15## 434 ± 69 呼吸（次/min） 78 ± 7 89 ± 8* 76.8 ± 9# 79 ± 8# 97 ± 15 应激反应（n） 0 1 0 1 5 中度以上出血（n） 0 0 0 0 2 与假手术组比较，*P < 0.05，**P < 0.01； 与模型组比较，#P < 0.05，##P < 0.01。 
下载: 导出CSV
表 2 Q-PCR引物序列
Table 2. Q-PCR primer sequences
序列 基因 产物长度 TATGGAATCCTGTGGCATC β actin-F 87 GTGTTGGCATAGAGGTCTT β actin--R GTCATAGCCACACTCAAG CINC1-F 75 TTGGACAATCTTCTGAACC CINC1-R CTCTCAAGGATGGTCAAG CINC2-F 81 GGCTCTTCAGTAACTTCT CINC2-R
下载: 导出CSV
表 3 灯盏花素对大鼠肺栓塞生命体征影响（
$\bar x \pm s $ ）Table 3. The effect of breviscapine on the vital signs of pulmonary embolism in rats （
$\bar x \pm s $ ）组别 假手术组 模型组 灯盏花素组 低分子肝素组 大鼠（只） 6 6 6 6 6 药物剂量 0 0 10 g/L 20 g/L 10.250 IU/L 心率（次/min） 356 ± 14 428 ± 31** 374 ± 25# 357 ± 19## 385 ± 33## 呼吸（次/min） 82 ± 9 94 ± 9* 76 ± 37 84 ± 10 83 ± 7# 与假手术组比较，*P < 0.05，**P < 0.01； 与模型组比较，#P < 0.05，##P < 0.01。
下载: 导出CSV
-
[1] Duffett L,Castellucci LA,Forgie MA. Pulmonary embolism:update on management and controversies[J]. BMJ,2020,370:2177. doi: 10.1136/bmj.m2177 [2] Watts JA,Lee YY,Gellar MA,et al. Proteomics of microparticles after experimental pulmonary embolism[J]. Thromb Res,2012,130(1):122-128. doi: 10.1016/j.thromres.2011.09.016 [3] Shibata F. The role of rat cytokine-induced neutrophil chemoattractants (CINCs) in inflamation[J]. Akugaku Zasshi,2002,122(4):263-268. doi: 10.1248/yakushi.122.263 [4] Zagorski J,Gellar M A,Obraztsova M,et al. Inhibition of CINC-1 decreases right ventricular damage caused by experimental pulmonary embolism in rats[J]. J Immunol,2007,179(11):7820-7826. doi: 10.4049/jimmunol.179.11.7820 [5] Li Y,Li S,Li D. Breviscapine Alleviates Cognitive Impairments Induced by Transient Cerebral Ischemia/Reperfusion through Its Anti-Inflammatory and Anti-Oxidant Properties in a Rat[J]. ACS Chem Neurosci,2020,11(24):4489-4498. [6] Wang C,Li Y,Gao S,et al. Breviscapine Injection Improves the Therapeutic Effect of Western Medicine on Angina Pectoris Patients[J]. PLoS One,2015,10(6):e0129969. doi: 10.1371/journal.pone.0129969 [7] Li Z,Lin H,Dong Z,et al. Protective effect of breviscapine in acute pulmonary embolism rats via regulation of MCP-1 and IL-13[J]. Journal of Cellular and Molecular Medicine,2018,22(12):6405-6407. doi: 10.1111/jcmm.13871 [8] Jarman AF,Mumma BE,Singh KS,et al. Crucial considerations:Sex differences in the epidemiology,diagnosis,treatment,and outcomes of acute pulmonary embolism in non-pregnant adult patients[J]. J Am Coll Emerg Physicians Open,2021,2(1):e12378. [9] Cohen AT,Katholing A,Rietbrock S,et al. Epidemiology of first and recurrent venous thromboembolism in patients with active cancer. A population-based cohort study[J]. Korean J Intern Med,2019,34(5):1125-1135. doi: 10.3904/kjim.2018.097 [10] Bircan HA,Alanoglu EG. Massive Pulmonary Embolism in a Patient with Heparin Induced Thrombocytopenia:Successful Treatment with Dabigatran[J]. Eurasian J Med,2016,48(1):65-68. doi: 10.5152/eurasianjmed.2015.95 [11] Condliffe R,Elliot CA,Hughes RJ,et al. Management dilemmas in acute pulmonary embolism[J]. Thorax,2014,69(2):174-180. doi: 10.1136/thoraxjnl-2013-204667 [12] Adegunsoye A,Balachandran J. Inflammatory response mechanisms exacerbating hypoxemia in coexistent pulmonary fibrosis and sleep apnea[J]. Mediators Inflamm,2015,2015:510105. [13] Watts JA,Zagorski J,Gellar MA et al. Cardiac inflammation contributes to right ventricular dysfunction following experimental pulmonary embolism in rats[J]. J Mol Cell Cardiol,2006,41(2):296-307. doi: 10.1016/j.yjmcc.2006.05.011 [14] Pilarczyk K,Renner J,Haake N. Acute right heart failure on the intensive care unit:Pathophysiology,monitoring and management[J]. Med Klin Intensivmed Notfmed,2019,114(6):567-588. doi: 10.1007/s00063-019-0603-6 -
点击查看大图
计量
- 文章访问数: 2430
- HTML全文浏览量: 1742
- PDF下载量: 23
- 被引次数: 0
