Effect of acupuncture in intervening heroin-induced brain damage via regulating ubiquitin-proteasome pathway

Abstract

Objective

To observe the effect of acupuncture in regulating ubiquitin-proteasome pathway (UPP), and discuss the action of acupuncture in intervening heroin-induced brain damage.

Methods

Thirty male Sprague-Dawley (SD) rats were divided into a control group, a model group and an acupuncture group by using the random number table. Rats in the model and acupuncture groups received intramuscular heroin injection for successive 8 d at a progressively increased dose. Afterwards, the injection was suspended for 5 d for withdrawal. The heroin relapse rat model was established by repeating the drug addiction and withdrawal process for 3 times. The control group followed the step of the model establishment, but was given intramuscular injection of normal saline at the stage of addiction and no intervention at the stage of withdrawal; the model group was given intramuscular heroin injection at a progressively increased dose at the addiction stage and no intervention at the withdrawal stage; the acupuncture group was dealt in the same way as the model group at the addiction stage, but received acupuncture at Baihui (GV 20) and Dazhui (GV 14) at the withdrawal stage, with the needles retained for 30 min each time, 1 session a day, for successive 5 d. On the 39th day, brain tissues were extracted from the hippocampus and ventral tegmental area (VTA) of the three groups of rats. The apoptosis of brain nerve cells was detected by using terminal deoxynucleotidyl transferase-mediated nick and labeling (TUNEL). The mRNA and protein expressions of ubiquitin (Ub), ubiquitin protein ligase (E3) and 26S were examined by immunohistochemistry and quantitative real-time polymerase chain reaction (RT-qPCR).

Results

Compared with the model group, rat’s hippocampus and VTA in the acupuncture group showed significantly fewer cells positively stained by TUNEL staining (P<0.01), and its mRNA and protein expressions of Ub, E3, 26S were significantly lower (P<0.01).

Conclusion

Reducing nerve cell apoptosis and regulating the mRNA and protein expressions of Ub, E3 and 26S in rat’s hippocampus and VTA are possibly one of the action mechanisms of acupuncture in intervening heroin-induced brain damage.

摘要

目的

观察针刺对泛素-蛋白酶体途径(UPP)的调控作用, 探讨针刺干预海洛因脑损伤的作用。

方法

将30 只雄性Sprague-Dawley (SD)大鼠按随机数字表法分为对照组、 模型组和针刺组。 模型组和针刺组大鼠采用连续8 d 递增量肌肉注射海洛因, 染毒后停止肌肉注射海洛因5 d, 自然戒断。按染毒(成瘾)-脱毒的方法, 反复3个阶段, 建立海洛因复吸大鼠模型。 对照组按照建立海洛因复吸大鼠模型的周期, 在染毒期给予大鼠肌肉注射生理盐水, 在 脱毒期不给予任何治疗; 模型组在染毒期给予大鼠连续递增量肌肉注射海洛因, 在脱毒期不给予任何治疗; 针刺组在染毒期处理与模型组相同, 在脱毒期给予针刺百会和大椎治疗, 每次留针30 min, 每天1次, 连续5 d。于实验 第39 d取3组大鼠的海马、 中脑腹侧被盖区(VTA)脑组织。 运用脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)检测脑神经细胞的凋亡情况。 运用免疫组化法、 定量实时PCR (RT-qPCR)法检测泛素(Ub)、 泛素蛋白连接酶 (E3)、 26S mRNA和蛋白的表达。

结果

与模型组相比, 针刺组大鼠海马、 VTA中TUNEL染色阳性细胞数显著减少 (P<0.01), Ub、 E3、 26S mRNA和蛋白表达表达明显减少(P<0.01)。

结论

减少神经细胞凋亡, 调节大鼠海马、 VTA 区的Ub、 E3、 26S mRNA和蛋白的表达, 可能是针刺干预海洛因脑损伤的作用机制之一。