针刺背俞穴对失眠模型大鼠脑组织磷脂酰肌醇3-激酶和蛋白激酶B表达的影响

Effect of Acupuncture at Back-Shu Points on Expression of PI3K and Akt in Brain Tissue of Insomnia Model Rats

目的 观察针刺背俞穴对失眠模型大鼠脑组织磷脂酰肌醇3-激酶(PI3K)和蛋白激酶B(Akt)表达的影响,探讨针刺改善失眠症状的可能机制。方法 将20只SPF级健康雄性SD大鼠随机分为空白组、模型组、针刺组和西药组,每组5只。除空白组外,其余3组采用腹腔注射对氯苯丙氨酸(PCPA)混悬液诱导失眠模型。造模2 d后,针刺组予以针刺双侧“肺俞”“脾俞”“心俞”“肝俞”“肾俞”,留针10 min,每日1次,治疗7 d;西药组予以艾司唑仑水溶液(0.08 mg/mL)灌胃治疗(剂量为0.558 mg/kg),每日1次,治疗7 d;其余各组大鼠只被捉抓和固定,不予治疗。观察各组大鼠治疗前后一般状态的变化,采用Western blot和实时荧光PCR法检测大鼠脑组织中PI3K、Akt蛋白水平和mRNA的表达情况。结果 与空白组比较,模型组大鼠一般状态较差,昼夜节律消失,白天活动增加,睡眠潜伏期延长,睡眠时间缩短,脑组织中PI3K、Akt蛋白和mRNA表达量显著降低(P<0.01);与模型组比较,针刺组和西药组大鼠一般状态有所改善,睡眠潜伏期缩短,睡眠时间延长,脑组织中PI3K、Akt蛋白和mRNA的表达均显著升高(P<0.05);与西药组比较,针刺组大鼠一般状况及脑组织中PI3K、Akt的mRNA升高较明显(P<0.05),差异存在统计学意义。结论 针刺背俞穴可升高失眠模型大鼠脑组织中PI3K、Akt蛋白及mRNA的表达,激活PI3K/Akt信号通路的活性,保护神经细胞,从而改善大鼠失眠情况。

Objective The effect of acupuncture at back-Shu points on the expression of PI3K and Akt in brain tissue of insomnia model rats were observed to explore the possible mechanism of acupuncture on improving insomnia symptoms.Methods Twenty SPF healthy male SD rats were randomly divided into blank group,model group,acupuncture group and Western medicine group,with 5 rats in each group.In addition to the blank group,the insomnia model was induced by intraperitoneal injection of PCPA suspension in the 3 groups.After 2 days of modeling,the acupuncture group was treated with acupuncture at bilateral“Feishu(BL13)”“Pishu(BL20)”“Xinshu(BL15)”“Ganshu(BL18)”and“Shenshu(BL23)”for 10 minutes and for 7 days;the Western medicine group was treated with estazolam solution(0.08 mg/mL)by gavage(the dose was 0.558 mg/kg)once a day for 7 days.Grouped rats were only caught and fixed without treatment.To observe the changes of the general state of the rats in each group before and after treatment.The protein and mRNA expressions of PI3K and Akt in brain tissue were detected by Western Blot and real-time PCR.Results Compared with the blank group,the rats in the model group had poor general state,disappeared circadian rhythm,increased daytime activity,prolonged sleep latency,shortened sleep time,and significantly decreased PI3K,Akt protein and mRNA expression in brain tissue(P<0.01).Compared with the model group,the general state of rats in the acupuncture group and the Western medicine group was improved,the sleep latency was shortened,the sleep time was prolonged,and the expressions of PI3K,Akt protein and mRNA in the brain tissue were significantly increased(P<0.05).Compared with the Western medicine group,the general condition of rats in the acupuncture group and the mRNA levels of PI3K and Akt in the brain tissue were significantly increased(P<0.05),and the difference was statistically significant.Conclusion Acupuncture at back-Shu points can increase the expression of PI3K and Akt protein and mRNA in brain tissue of insomnia model rats,activate the activity of PI3K/Akt signaling pathway,protect nerve cells,and improve insomnia in rats.