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

Mechanism of Acupuncture Combined with Jing Tong Kang in Reducing CAS in Rats through PI3K/Akt/NF-κB Signaling Pathway

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45只随机分为模型组、假手术组、补阳还五汤组(简称原方组)、颈通康组与针刺结合颈通康组(简称针药组),每组9只。模型组、原方组、颈通康组与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灌胃给药;颈通康组予颈通康灌胃;针药组予颈通康灌胃,同时进行针刺;假手术组与模型组则使用等量生理盐水灌胃;2次/d,各组均连续给药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与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;应用全自动生化分析仪测定模型大鼠颈动脉组织中总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)水平。结果:与假手术组比较,模型组TC、TG、LDL-C、PI3K mRNA、Akt mRNA、NF-κB mRNA、PI3K、Akt与NF-κB显著升高,差异有统计学意义(P<0.01),HDL-C显著降低,差异具有统计学意义(P<0.01)。与模型组比较,原方组TG、LDL-C、Akt mRNA、PI3K、Akt及NF-κB显著降低,差异具有统计学意义(P<0.01),PI3K mRNA、NF-κB mRNA降低,差异具有统计学意义(P<0.05),HDL-C显著升高,差异具有统计学意义(P<0.01),TC无统计学意义;颈通康组TC、TG、LDL-C、PI3K mRNA、Akt mRNA、NF-κB mRNA、PI3K、Akt及NF-κB显著降低,差异具有统计学意义(P<0.01),HDL-C显著升高,差异具有统计学意义(P<0.01);针药组TC、TG、LDL-C、PI3K mRNA、Akt mRNA、NF-κB mRNA、PI3K、Akt及NF-κB显著降低,差异具有统计学意义(P<0.01),HDL-C显著升高,差异具有统计学意义(P<0.01)。与原方组比较,颈通康组TC、LDL-C、Akt mRNA、PI3K、Akt及NF-κB显著降低,差异具有统计学意义(P<0.01),HDL-C升高,差异具有统计学意义(P<0.05),TG、PI3K mRNA及NF-κB mRNA无统计学意义;针药组TC、TG、LDL-C、PI3K mRNA、Akt mRNA、NF-κB mRNA、PI3K、Akt及NF-κB显著降低,差异具有统计学意义(P<0.01),HDL-C显著升高,差异具有统计学意义(P<0.01)。与颈通康组比较,针药组TC、LDL-C、PI3K mRNA、NF-κB mRNA、PI3K、Akt及NF-κB显著降低,差异具有统计学意义(P<0.01),HDL-C显著升高,差异具有统计学意义(P<0.01),TG、Akt mRNA无统计学意义。结论:补阳还五汤、颈通康和针刺结合颈通康能够通过抑制PI3K/AKT/NF-κB信号通路减轻大鼠颈动脉狭窄,针刺结合颈通康疗效优于颈通康和补阳还五汤原方。

Objective:To explore the effect of acupuncture combined with Jing Tong Kang on severe carotid artery stenosis(CAS)in model rats and to investigate its mechanism in terms of the involved signaling pathway.Methods:45 SD male rats were randomly divided into the model group,the sham operation group,the Buyang Huanwu Decoction group,the Jing Tong Kang group,and the acupuncture plus medication group(acupuncture combined with Jing Tong Kang),with 9 rats in each group.The CAS model was established by carotid artery clamp combined with vitamin D3injection and high-fat diet.Although the neck was incised,the common carotid artery was not clamped in the sham operation group,and the rats in this group were fed with common rat feed.After successful modeling,the Buyang Huangwu Decoction group was given Buyang Huanwu Decoction by intragastric administration;the Jing Tong Kang group was administered with Jing Tong Kang by intragastric administration;the acupuncture plus medication group was treated with acupuncture and Jing Tong Kang.The sham operation group and the model group were given same amount of normal saline by gavage,2 times/d.After four weeks of the intervention,Western Blot was used to detect and analyze the protein expressions of PI3K,AKT and NF-κB in the carotid artery tissues;PCR was used to analyze the mRNA expressions of PI3K,AKT and NF-κB in the carotid artery tissues;the levels of total cholesterol(TC),triglycerides(TG),low density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C)in carotid artery were measured by automatic biochemical analyzer.Results:Compared with those in the sham operation group,TC,TG,LDL-C,PI3K mRNA,Akt mRNA,NF-κB mRNA,PI3K,Akt and NF-κB increased significantly(P<0.01),HDL-C decreased significantly in the model group(P<0.01).Compared with those in the model group,TG,LDL-C,Akt mRNA,PI3K,Akt and NF-κB decreased significantly(P<0.01),PI3K mRNA and NF-κB mRNA decreased(P<0.05),HDL-C increased significantly(P<0.01),but TC had no statistical significance in the Buyang Huanwu Decoction group;TC,TG,LDL-C,PI3K mRNA,Akt mRNA,NF-κB mRNA,PI3K,Akt and NF-κB decreased significantly(P<0.01),HDL-C increased significantly in the Jing Tong Kang group(P<0.01);TC,TG,LDL-C,PI3K mRNA,Akt mRNA,NF-κB mRNA,PI3K,Akt and NF-κB decreased significantly(P<0.01),and HDL-C increased significantly in the acupuncture plus medication group(P<0.01).Compared with those in the Buyang Huanwu Decoction group,TC,LDL-C,Akt mRNA,PI3K,Akt and NF-κB decreased significantly(P<0.01),HDL-C increased(P<0.05),TG,PI3K mRNA,NF-κB mRNA were not statistically significant in the Jing Tong Kang group;TC,TG,LDL-C,PI3K mRNA,Akt mRNA,NF-κB mRNA,PI3K,Akt and NF-κB decreased significantly(P<0.01),and HDL-C increased significantly in the acupuncture plus medication group(P<0.01).Compared with those in the Jing Tong Kang group,TC,LDL-C,PI3K mRNA,NF-κB mRNA,PI3K,Akt and NF-κB decreased significantly(P<0.01),HDL-C increased significantly(P<0.01),but TG and Akt mRNA had no statistical significance in the acupuncture plus medication group.Conclusion:Buyang Huanwu Decoction,Jing Tong Kang,and acupuncture combined with Jing Tong Kang can reduce CAS in rats by inhibiting PI3K/AKT/NF-κB signaling pathway.Among them,the efficacy of acupuncture combined with Jing Tong Kang is better than that of Jing Tong Kang alone and Buyang Huanwu Decoction.