复方麝香注射液联合神经节苷脂对弥散性轴索损伤大鼠神经元凋亡的影响

目的:探讨复方麝香注射液联合神经节苷脂对弥散性轴索损伤(DAI)大鼠神经元凋亡及神经丝蛋白200(NF200)、生长相关蛋白43(GAP-43)、凋亡相关蛋白激酶1(DAPK1)表达的影响。方法:采用大鼠头颅瞬间旋转损伤装置建立DAI大鼠模型。将成功建立的45只DAI大鼠随机分为模型组、实验组和联合组,每组15只;另选择15只正常大鼠作为对照组。实验组大鼠腹腔注射单唾液酸四己糖神经节苷脂钠30 mg/(kg·d)],联合组大鼠腹腔注射单唾液酸四己糖神经节苷脂钠30 mg/(kg·d)]+复方麝香注射液2 mL/(kg·d)],对照组与模型组腹腔注射等量生理盐水,连续干预7 d。采用改良神经功能缺损评分(mNSS)法评估各组大鼠的神经功能缺损情况;采用苏木精-伊红(HE)染色观察大鼠脑组织病理学变化;采用TdT介导的dUTP缺口末端标记(TUNEL)检测各组大鼠神经元凋亡情况;采用蛋白质免疫印迹(Wb)法检测大鼠脑组织NF200、GAP-43与DAPK1蛋白表达。结果:干预第1~7天,模型组、联合组与实验组大鼠mNSS评分均逐渐降低;与对照组比较,模型组大鼠第1~7天的mNSS评分均明显升高,但联合组与实验组低于模型组,联合组低于实验组(P<0.05);与对照组比较,模型组大鼠神经元凋亡指数及脑组织DAPK1蛋白相对表达量均明显升高,且联合组与实验组低于模型组,联合组低于实验组(P<0.05);NF200、GAP-43蛋白相对表达量明显降低,且联合组与实验组高于模型组,联合组高于实验组(P<0.05)。结论:复方麝香注射液联合神经节苷脂治疗DAI可改善大鼠神经功能缺损情况,抑制神经元凋亡,改善神经功能,其作用机制可能与上调脑组织NF200、GAP-43蛋白表达,下调DAPK1蛋白表达相关,且两药联合使用效果更佳。

Effects of Compound Shexiang Injection Combined with Ganglioside on Neuronal Apoptosis in Rats with Diffuse Axonal Injury

To explore effects of Compound Shexiang Injection combined with ganglioside on neuronal apoptosis and expressions of neurofilament 200(NF200),growth-associated protein 43(GAP-43)and death-associated protein kinase 1(DAPK1)in rats with diffuse axonal injury(DAI).Methods:DAI rat models were established using a rat skull transient rotation injury device.Forty-five DAI rats successfully established were randomly divided into model group,test group and combined group,with 15 rats in each group.Another 15 normal rats were selected as control group.Rats in the test group were intraperitoneally injected with monosialotetrahexosyl ganglioside sodium30 mg/(kg·d)],rats in the combined group were injected intraperitoneally with monosialotetrahexosyl ganglioside sodium(30 mg·kg-1·d-1)+Compound Shexiang Injection(2 mL·kg-1·d-1),and the model and control groups were injected intraperitoneally with the same amount of normal saline,for 7 d.Modified neurological severity score(mNSS)method was used to evaluate neurological deficits in each group of rats.Hematoxylin-eosin(HE)staining was used to observe pathological changes of brain tissue in rats.TdT-mediated dUTP Notch End Labeling(TUNEL)was used to detect neuronal apoptosis in each group of rats.Protein expressions of NF200,GAP-43 and DAPK1 in rat brain tissue were detected by Western blot(Wb)method.Results:From the 1st to 7th d of the intervention,the mNSS scores of rats in the model group,the combined group and the test group were gradually decreased.Compared with the control group,the mNSS scores of the rats in the model group were significantly increased from the 1st to 7th d,but those in the combined group and the test group were lower than those in the model group,and those in the combined group were lower than those in the test group(P < 0.05).Compared with the control group,the neuronal apoptosis index and the relative expression of DAPK1 protein in brain tissue of rats in the model group were significantly increased,and those in the combined group and the test group were lower than those in the model group,and those in the combined group were lower than those in the test group(P < 0.05).The relative expressions of NF200 and GAP-43 proteins were significantly decreased,and those in the combined group and the test group were higher than those in the model group,and those in the combined group were higher than those in the test group(P < 0.05).Conclusion:Compound Shexiang Injection combined with ganglioside in the treatment of DAI can improve neurological deficits in rats,inhibit neuronal apoptosis,and improve neural function.Its mechanism of action may be related to up-regulation of NF200 and GAP-43 protein expressions in brain tissue and down-regulation of DAPK1 protein expression,and the combined use of the 2 drugs is more effective.