针刺结合颈通康通过PI3K/Akt/NF-κB信号通路减轻大鼠颈动脉狭窄机制研究

周海纯; 佟春雪; 宋志强; 韩晓庆; 刘艳英

文章摘要

针刺结合颈通康通过PI3K/Akt/NF-κB信号通路减轻大鼠颈动脉狭窄机制研究

Study on the mechanism of acupuncture combined with Jingtongkang in reducing carotid artery stenosis in rats through PI3K/Akt/NF-κB signaling pathway

投稿时间：2022-04-08 录用日期：2022-05-26

DOI：

中文关键词: 关键词:针刺、补阳还五汤、PI3K/Akt/NF-κB、颈动脉狭窄

英文关键词: Key words: acupuncture Buyang Huanwu Decoction PI3K/Akt/NF-κB carotid artery stenosis

基金项目:

作者	单位	邮编
周海纯	黑龙江中医药大学附属第二医院哈南分院内五科	150040
佟春雪	黑龙江中医药大学附属第二医院哈南分院内五科	150040
宋志强	黑龙江中医药大学附属第二医院哈南分院内五科	150040
韩晓庆	黑龙江中医药大学附属第二医院哈南分院内五科	150040
刘艳英^*	黑龙江中医药大学附属第二医院哈南分院内五科	150040

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中文摘要:

摘要：目的：探究针刺结合颈通康对重度颈动脉狭窄模型大鼠的影响及所涉及的信号通路机制。方法：将SD雄性大鼠（200g±20g）45只按随机数字表法随机分为模型组、假手术组、补阳还五汤组(简称原方组)、颈通康组、针刺结合颈通康组(简称针药组)，每组9只。模型组、原方组、颈通康组、针药组均采用颈动脉钳夹联合维生素D3注射、高脂饮食构建颈动脉粥样硬化性狭窄大鼠模型，假手术组给以普通饲料喂养，颈部虽切开但不进行颈总动脉钳夹术。待造模成功后，原方组予补阳还五汤原方13.3mg/kg灌胃给药；颈通康组予颈通康13.3mg/kg灌胃给药；针药组予颈通康13.3mg/kg灌胃给药，同时进行针刺大鼠足三里、丰隆、内关、人迎；假手术组与模型组则使用等量生理盐水灌胃，2次/d，各个组均连续给药4周。干预结束后，应用Western Blot检测分析模型大鼠颈动脉组织中PI3K、AKT、NF-κB蛋白表达情况；应用PCR分析模型大鼠颈动脉组织中PI3K、AKT、NF-κB mRNA表达情况。结果：经qRT-PCR检测分析，与假手术组相比，模型组、原方组大鼠PI3K、AKT、NF-κB mRNA表达均有明显升高（p<0.01），颈通康组、针药组大鼠PI3K、NF-κB mRNA表达均显著升高（p<0.01）而AKT mRNA表达无统计学差异；与模型组比较，原方组大鼠PI3K、NF-κB mRNA表达下降（p＜0.05），原方组大鼠AKT mRNA表达显著下降（p＜0.01），颈通康组、针药组大鼠PI3K、AKT、NF-κB mRNA表达均显著下降（p＜0.01）；与原方组相比较，颈通康组大鼠仅AKT mRNA表达均显著降低（p＜0.01），针药组大鼠PI3K、AKT、NF-κB mRNA表达均显著降低（p＜0.01）；与颈通康组做比较，针药组大鼠PI3K、NF-κB mRNA表达显著降低（p＜0.01），而AKT mRNA表达无统计学差异。经Western Blot检测分析，与假手术组进行比较，模型组、原方组、颈通康组、针药组大鼠AKT、PI3K、NF-κB蛋白表达显著升高（p＜0.01）；与模型组比较，原方组、颈通康组、针药组大鼠AKT、PI3K、NF-κB蛋白表达显著下降（p＜0.01）；与原方组做对比，颈通康组、针药组大鼠AKT、PI3K、NF-κB蛋白表达下降明显（p＜0.01）；与颈通康组比较，针药组大鼠AKT、PI3K、NF-κB蛋白表达有显著减少（p＜0.01）。结论：补阳还五汤、颈通康、针刺结合颈通康至少能够通过抑制PI3K/AKT/NF-κB信号通路减轻大鼠颈动脉狭窄。其中，针刺结合颈通康疗效优于颈通康优于补阳还五汤原方。

英文摘要:

Abstract:Objective:To explore the effect of acupuncture combined with Jingtongkang on severe carotid stenosis model rats and the involved signaling pathway mechanism.Methods:45 SD male rats (200g±20g) were randomly divided into model group, sham operation group, Buyang Huanwu decoction group (referred to as Yuanfang group), Jingtongkang group, and acupuncture combined with Jingtongkang group (referred to as acupuncture and medicine group)according to random number table,9 rats in each group. The model group, Yuanfang group, Jingtongkang group, and acupuncture and medicine group were all used carotid artery clamp combined with vitamin D3 injection and high-fat diet to establish a rat model of carotid atherosclerotic stenosis. Although the neck was incised, the common carotid artery was not clamped.After successful modeling, the Yuanfang group was given Buyang Huanwu Decoction 13.3mg/kg by intragastric administration; the Jingtongkang group was administered with Jingtongkang 13.3mg/kg by intragastric administration; the acupuncture and medicine group was administered with Jingtongkang 13.3mg/kg and the rats were given acupuncture at Zusanli, Fenglong, Neiguan and Renying at the same time; the sham-operated group and the model group were given the same amount of normal saline by gavage, 2 times/d. All groups were given continuous administration for 4 weeks.After the intervention, Western Blot was used to detect and analyze the protein expressions of PI3K, AKT and NF-κB in the carotid artery tissue of the model rats; PCR was used to analyze the mRNA expressions of PI3K, AKT and NF-κB in the carotid artery tissue of the model rats.Results:By qRT-PCR analysis ,compared with the sham-operated group, the PI3K, AKT and NF-κB mRNA expressions in the model group and the Yuanfang group were significantly increased (p<0.01), and in the Jingtongkang group and the acupuncture and medicine group, the expressions of PI3K and NF-κB mRNA were significantly increased (p<0.01), but there was no statistical difference in the expression of AKT mRNA;Compared with the model group, the expression of PI3K and NF-κB mRNA in the Yuanfang group decreased (p<0.05), and the AKT mRNA expression in the Yuanfang group decreased significantly (p<0.01). In the Jingtongkang group and acupuncture and medicine group ,the mRNA expressions of PI3K, AKT and NF-κB were all significantly decreased (p<0.01);Compared with the Yuanfang group, only the mRNA expression of AKT in the Jingtongkang group was significantly decreased (p<0.01), and the mRNA expressions of PI3K, AKT and NF-κB were significantly decreased in the acupuncture and medicine group (p<0.01);Compared with the Jingtongkang group, the mRNA expressions of PI3K and NF-κB in the acupuncture and medicine group were significantly decreased (p<0.01), while the AKT mRNA expression had no statistical difference.By Western Blot analysis, compared with the sham operation group, the expression of AKT, PI3K and NF-κB proteins in the model group, Yuanfang group, Jingtongkang group, and acupuncture and medicinde group were significantly increased (p<0.01);Compared with the model group, the protein expressions of AKT, PI3K and NF-κB in the Yuanfang group, the Jingtongkang group and the acupuncture and medicine group were significantly decreased (p<0.01);Compared with the Yuanfang group, the proteins expressions of AKT, PI3K and NF-κB in the Jingtongkang group and the acupuncture and medicine group were significantly decreased (p<0.01);Compared with the Jingtongkang group,the expression of AKT, PI3K and NF-κB proteins in the acupuncture and medicine group were significantly decreased(p<0.01).Conclusion:Buyang Huanwu Decoction,Jingtongkang,acupuncture combined with Jingtongkang can at least reduce carotid artery stenosis in rats by inhibiting PI3K/AKT/NF-κB signaling pathway. Among them, the efficacy of acupuncture combined with Jingtongkang is better than that of the original formula of Buyang Huanwu Decoction.

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