红景天苷调控AMPK/Drp1信号通路对急性脑出血大鼠的神经功能的保护机制

Neuroprotection and mechanism of AMPK/Drp1 signaling pathway modulation by salidroside in rats with acute cerebral hemorrhage

目的研究红景天苷(salidroside,Sal)对急性脑出血(intracerebral hemorrhage,ICH)大鼠的神经功能保护作用及其机制。方法根据Rosenberg法改良造大鼠ICH神经损伤模型。按照随机性原则将SD大鼠随机分为假手术组(Sham)、模型组(Model)、Sal低、中、高(50、100、200 mg·kg^(-1))、AMPK激动剂(AICAR)组、阳性药(醒脑静)组,每组各10只。于造模结束1 h后腹腔注射各剂量Sal,连续1周,AMPK激动剂组腹腔注射AICAR(200 mg·kg^(-1)),阳性药组腹腔注射醒脑静注射液(1.8 mL·kg^(-1)),假手术组和模型组腹腔注射等量生理盐水。采用神经评分、脑含水量等检测各组大鼠的神经功能损伤;MitoSOX Red试剂盒检测各组大鼠脑组织ROS水平;ELISA测定血清炎性因子IL-6、IL-1β及TNF-α水平;TUNEL染色检测各组大鼠神经细胞凋亡情况;Western blot检测相关蛋白Bcl-2、Bax、cleaved caspase-3、AMPK/Drp1的表达。结果与Sham组相比,Model组大鼠神经功能损伤严重,出现明显脑水肿,脑组织内氧化损伤和炎症明显加重且神经细胞出现大量凋亡(P<0.05),Bcl-2、p-Drp1/Drp1蛋白表达明显下调,Bax、cleaved caspase-3、p-AMPK/AMPK蛋白表达明显上调(P<0.05);与Model组相比,Sal各剂量治疗组、阳性药组和AICAR组大鼠神经功能损伤明显减轻,脑水肿情况得到缓解,脑组织内氧化损伤和炎症明显减轻且神经细胞凋亡被抑制(P<0.05),Bcl-2、p-AMPK/AMPK蛋白表达明显上调,Bax、cleaved caspase-3、p-Drp1/Drp1蛋白表达明显下调(P<0.05)。结论Sal能够有效的保护急性ICH大鼠的神经功能,其作用机制可能与AMPK/Drp1信号通路有关。

Aim To study the protective effect of salidroside(Sal)on neurological function in rats with acute cerebral hemorrhage,and to explore its mechanism.Methods The Rosenberg method was used to improve the neural injury model of rats with cerebral hemorrhage.SD rats were randomly divided into sham group(Sham),model group(Model),salidroside low dose group(50 mg·kg^(-1)),salidroside medium dose group(100 mg·kg^(-1)),salidroside high dose group(200 mg·kg^(-1))and AMPK agonist group(AICAR),with 10 rats in each group.One hour after the end of modeling,salidroside was intraperitoneally injected with each dose for one week.AMPK agonist group was intraperitoneally injected with AICAR(200 mg·kg^(-1)),while sham operation group and model group were intraperitoneally injected with the same amount of normal saline.Nerve score and brain water content were used to detect nerve function injury in each group.MitoSOX Red kit was used to detect reactive oxygen species in brain tissue of rats.Enzyme-labeled immunosorbent assay(ELISA)was used to determine the serum levels of inflammatory cytokines IL-6,IL-1βand TNF-α.Tunel staining was used to detect apoptosis of nerve cells in each group.The expressions of Bcl-2,Bax,cleaved caspase-3,and AMPK/Drp1 were detected by Western blot.Results Compared with the sham group,the model group and AICAR group had severe neurological function injury,cerebral edema,oxidative damage and inflammation in brain tissue were significantly aggravated,and apoptosis appeared in a large number of nerve cells(P<0.05).Western blot results showed that the protein expressions of Bcl-2 and P-DRP1/Drp1 were significantly down-regulated,and the protein expressions of Bax,cleaved caspase-3 and P-AMPK/AMPK were significantly up-regulated(P<0.05).Compared with the model group,neurological function injury and brain edema were significantly alleviated in the salidroside treatment groups,oxidative damage and inflammation in brain tissue were significantly alleviated,and nerve cell apoptosis was inhibited(P<0.05).Western blot results showed that Bcl-2 and P-AMPK/AMPK protein expressions were significantly up-regulated.The protein expressions of Bax,cleaved caspase-3,and P-DRP1/Drp1 significantly decreased(P<0.05).Conclusion Our study shows that salidroside can effectively protect the neurological function of rats with acute cerebral hemorrhage,and its mechanism may be related to AMPK/Drp1 signaling pathway.