Breviscapine Regulates the mRNA and Protein Levels of CINC-1 and CINC-2 in the Right Ventricle of Rats with Acute Pulmonary Embolism
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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
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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。 
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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
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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。
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