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| 右美托咪定激活Akt通路对缺氧复氧肾小管上皮细胞凋亡的影响 |
| Effect of dexmedetomidine activating Akt Pathway on apoptosis of renal tubular epithelial cells after hypoxia reoxygen |
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| DOI：10.12089/jca.2020.09.014 |
| 中文关键词: 右美托咪定 缺氧复氧 肾小管上皮细胞 Akt通路 凋亡 |
| 英文关键词: Dexmedetomidine Hypoxia reoxygenation Renal tubular epithelial cells Akt pathway Apoptosis |
| 基金项目: |
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| 摘要点击次数: 1887 |
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| 中文摘要: |
目的 观察右美托咪定对缺氧复氧诱导肾小管上皮细胞凋亡的作用，并探讨其作用机制。
方法 将人肾小管上皮细胞分五组干预：对照组(C组)，常氧环境中培养28 h；缺氧复氧组(HR组)，低氧环境中培养24 h后常氧环境中培养4 h；右美托咪定处理组(D组)，复制缺氧复氧模型前以右美托咪定处理2 h；Akt阻断剂Uprosertib处理组(U组)，复制缺氧复氧模型前以Akt阻断剂Uprosertib处理1 h；右美托咪定复合Akt阻断剂Uprosertib处理组(DU组)，复制缺氧复氧模型前先以Akt阻断剂Uprosertib处理1 h，再以右美托咪定处理2 h。采用Ellsa法检测细胞上清中TNF-ɑ、IL-6、IL-1β浓度，MTT法检测细胞存活率，流式细胞仪检测细胞凋亡率，Western blot法检测Bcl-2、Bax、caspase-3、caspase-9、p-Akt蛋白含量。
结果 与C组比较，HR组、D组、U组和DU组TNF-ɑ、IL-6、IL-1β浓度、细胞凋亡率、Bax、caspase-3、caspase-9蛋白含量明显升高(P<0.05)，细胞存活率、Bcl-2、p-Akt蛋白含量明显降低(P<0.05)。与HR组比较，D组和DU组TNF-ɑ、IL-6、IL-1β浓度、细胞凋亡率、Bax、caspase-3、caspase-9蛋白含量明显降低(P<0.05)，细胞存活率、Bcl-2、p-Akt蛋白含量明显升高(P<0.05)；U组TNF-ɑ、IL-6、IL-1β浓度、细胞凋亡率、Bax、caspase-3、caspase-9蛋白含量明显升高(P<0.05)，细胞存活率、Bcl-2、p-Akt蛋白含量明显降低(P<0.05)。与D组比较，U组和DU组TNF-ɑ、IL-6、IL-1β浓度、细胞凋亡率、Bax、caspase-3、caspase-9蛋白含量明显升高(P<0.05)，细胞存活率、Bcl-2、p-Akt蛋白含量明显降低(P<0.05)。与U组比较，DU组TNF-ɑ、IL-6、IL-1β浓度、细胞凋亡率、Bax、caspase-3、caspase-9蛋白含量明显降低(P<0.05)，细胞存活率、Bcl-2、p-Akt蛋白含量明显升高(P<0.05)。
结论 右美托咪定可通过激活Akt信号通路，抑制缺氧复氧造成的肾小管上皮细胞凋亡，发挥保护作用。 |
| 英文摘要: |
Objective To observe the effect of dexmedetomidine on apoptosis of renal tubular epithelial cells induced by hypoxia and reoxygenation.
Methods Human renal tubular epithelial cells were divided into 5 groups for intervention: control group (group C), cultured in normoxia for 28 h; hypoxia reoxygenation group (group HR), cultured in hypoxic environment for 24 h and then cultured in normoxia for 4 h; dexmedetomidine treatment group (group D), treated with dexmedetomidine for 2 h before replicating the hypoxia reoxygenation model, Akt blocker Uprosertib treatment group (group U), blocked with Akt 1 h before the replication of the hypoxia-reoxygenation model Uprosertib treatment 1 h; dexmedetomidine combined with Akt blocker Uprosertib treatment group (group DU), before copying the hypoxic-reoxygenation model, the Akt blocker Uprosertib treatment 1h, then dexmedetomidine treatment 2 h. Ellsa method was used to detect the concentration of TNF-ɑ, IL-1β and IL-6 in cell supernatant, MTT method was used to detect cell survival rate, flow cytometry was used to detect cell apoptosis rate, Western blot was used to detect Bcl-2, Bax, caspase 3. Protein content of caspase-9 and p-Akt.
Results Compared with group C, the concentrations of TNF-α, IL-6, IL-1 β ， the apoptosis rate, the protein contents of Bax, caspase-3 and caspase-9 were significantly increased (P < 0.05), the cell survival rate, the protein contents of Bcl-2 and p-Akt were significantly decreased in groups HR, D, U and DU (P < 0.05). Compared with group HR, the concentrations of TNF-α, IL-6, IL-1 β, the apoptosis rate, the protein contents of Bax, caspase-3 and caspase-9 were significantly decreased (P < 0.05), the cell survival rate, the protein contents of Bcl-2 and p-Akt were significantly increased in group D and DU (P < 0.05), the concentrations of TNF-α, IL-6, IL-1 β， the apoptosis rate, the protein contents of Bax, caspase-3 and caspase-9 were significantly increased (P < 0.05), the cell survival rate, the protein contents of Bcl-2 and p-Akt were significantly decreased in group U (P < 0.05). Compared with group D, the concentrations of TNF-α, IL-6, IL-1 β ， the apoptosis rate, the protein contents of Bax, caspase-3 and caspase-9 increased were significantly increased (P < 0.05), the cell survival rate, the protein contents of Bcl-2 and p-Akt were significantly decreased in groups U and DU (P < 0.05). Compared with group U, the concentrations of TNF-α, IL-6, IL-1 β, the apoptosis rate, the protein contents of Bax, caspase-3 and caspase-9 were significantly decreased (P < 0.05), the cell survival rate, the protein contents of Bcl-2 and p-Akt were significantly increased in group DU (P < 0.05).
Conclusion Dexmedetomidine can protect renal tubular epithelial cells from apoptosis induced by hypoxia and reoxygenation by activating Akt signaling pathway. |
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