| 代楚行,陈俊,王壹民,等.基于CXCL12/CXCR4生物轴探讨清心饮抑制CVB3感染后CMVECs发生EndMT的作用[J].浙江中医药大学学报,2022,46(7):699-707, 727. |
| 基于CXCL12/CXCR4生物轴探讨清心饮抑制CVB3感染后CMVECs发生EndMT的作用 |
| Observation on the Action of Qingxinyin Inhibiting EndMT in CVB3-infected CMVECs Based on CXCL12/CXCR4 Biological Axis |
| DOI：10.16466/j.issn1005-5509.2022.07.001 |
| 中文关键词: 清心饮 心脏微血管内皮细胞 柯萨奇B3病毒 内皮间充质转分化 CXCL12/CXCR4生物轴 |
| 英文关键词: Qingxinyin CMVECs CVB3 EndMT CXCL12/CXCR4 biological axis |
| 基金项目:国家自然科学基金面上项目(81873247) |
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| 摘要点击次数: 2031 |
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| 中文摘要: |
| [目的] 探讨趋化因子12(chemokine 12，CXCL12)/趋化因子受体4(chemokine receptor 4，CXCR4)生物轴在清心饮含药血清抑制柯萨奇B3病毒(Coxsackie virus B3，CVB3)感染后心脏微血管内皮细胞(cardiac microvascular endothelial cells，CMVECs)发生内皮间充质转分化(endothelial-mesenchymal transition，EndMT)过程中的作用。[方法] 取生长状态良好的CMVECs设计为正常对照组、模型组、CXCR4拮抗剂组、清心饮组和清心饮+CXCR4拮抗剂组。除正常对照组用含10%胎牛血清(fetal bovine serum，FBS)的杜尔伯克改良伊格尔培养基(Dulbecco‘s modification of Eaglews medium，DMEM)培养外，其余各组CMVECs先用CVB3培养基感染2 h，后弃去CVB3培养基，改为普通培养。通过细胞计数试剂(cell counting kit-8，CCK8)检测各组细胞活力，免疫荧光(immunofluorescence，IF)、免疫印迹和实时荧光定量聚合酶链式反应(Real-time quantitative polymerase chain reaction，Real-time qPCR)等实验方法检测内皮细胞标志物血小板-内皮细胞黏附分子(platelet endothelial cell adhesion molecule-1，PECAM-1/CD31)、间充质细胞标志物α-平滑肌肌动蛋白(α-smooth muscle actin，α-SMA)、CXCL12和CXCR4的蛋白及基因表达。[结果] 与正常对照组比较，模型组CD31表达下降(P＜0.01)，α-SMA、CXCL12和CXCR4表达增强(P＜0.01)。与模型组比较，CXCR4拮抗剂组、清心饮组CD31表达增强(P＜0.01)，α-SMA和CXCR4表达下降(P＜0.01)，CXCR4拮抗剂组CXCL12表达差异无统计学意义(P＞0.05)，清心饮组CXCL12表达下降(P＜0.05)。与清心饮组比较，清心饮+CXCR4拮抗剂组CD31的表达增强(P＜0.05)，α-SMA、CXCL12和CXCR4表达下降(P＜0.01，P＜0.05)。[结论] CXCL12/CXCR4生物轴参与了CVB3感染后CMVECs发生EndMT的过程，拮抗CXCL12/CXCR4生物轴能够明显增强清心饮对该过程的抑制。 |
| 英文摘要: |
| [Objective] To explore the intervention effect of Qingxinyin pharmacologic serum in inhibiting the endothelial-mesenchymal transition(EndMT) in Coxsackie virus B3(CVB3) infected cardiac microvascular endothelial cells(CMVECs) via regulating chemokine 12/ chemokine receptor 4(CXCL12/CXCR4) biological axis. [Methods] CMVECs in good growth status were taken， and according to the experimental purpose， normal control group， model group， CXCR4 antagonist group， Qingxinyin group and Qingxinyin+CXCR4 antagonist group were designed. Except for normal control group， which was cultured with Dulbecco‘s modification of Eagle‘s medium(DMEM) containing 10% fetal bovine serum(FBS)， the other groups were infected CMVECs with CVB3 culture medium for 2 hours， and abandoned CVB3 culture medium， and then continued to normal culture. The cell activity of each group was performed by cell counting kit-8(CCK8). The protein and gene expressions of the endothelial cell marker platelet endothelial cell adhesion molecule-1(PECAM-1/CD31)， the mesenchymal cell marker α-smooth muscle actin(α-SMA)， CXCL12 and CXCR4 were detected with immunofluorescence(IF)， Western blot and Real-time quantitative polymerase chain reaction(Real-time qPCR). [Results] Compared with normal control group， the expression of CD31 was decreased(P＜0.01)， and the expressions of α-SMA， CXCL12 and CXCR4 were increased in model group(P＜0.01). Compared with model group， the expression of CD31 was increased(P＜0.01)， and the expressions of α-SMA and CXCR4 were reduced in CXCR4 antagonist group and Qingxinyin group(P＜0.01)， there was no statistically significant expression of CXCL12 in CXCR4 antagonist group(P＞0.05)， the expression of CXCL12 was decreased in Qingxinyin group(P＜0.05). Compared with Qingxinyin group， the expression of CD31 was increased(P＜0.05)， and the expressions of α-SMA， CXCL12 and CXCR4 were reduced in Qingxinyin+CXCR4 antagonist group(P＜0.01， P＜0.05). [Conclusion] The CXCL12/CXCR4 biological axis is involved in the occurrence of EndMT in CVB3-infected CMVECs， restraining CXCL12/CXCR4 biological axis which would significantly increase the effect of inhibition. |
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