摘要
目的探究三七总皂苷对脑挫伤大鼠神经元凋亡的影响,分析其与腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)/过氧化物酶体增生物激活受体γ共激活因子1α(peroxisome proliferator activated receptor gamma co-activator 1α,PGC1α)通路的调控机制。方法从98只SD大鼠中随机选取16只为假手术组,剩余82只大鼠构建创伤性脑损伤(traumatic brain injury,TBI)模型,将模型建立成功的64只大鼠随机分为模型组、三七总皂苷组(26.25 mg/kg)、Compound C组(CC,AMPK抑制剂,0.25 mg/kg)和三七总皂苷+CC组(三七总皂苷26.25 mg/kg+CC 0.25 mg/kg),每组16只,造模后24 h进行药物处理,持续21 d;给药结束后,采用改良神经功能缺损评分(modified neurological deficit score,mNSS)量表评估各组大鼠神经功能;比色法检测各组大鼠脑组织超氧化物歧化酶(superoxide dismutase,SOD)和丙二醛(malondialdehyde,MDA)水平;苏木精伊红(hematoxylin eosin,HE)染色、βⅢ微管蛋白(neuron-specific classⅢbeta-tubulin,Tuj1)免疫荧光染色和末端脱氧核苷酸转移酶介导的dUTP原位切口末端标记(terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling,TUNEL)染色分别显示大鼠脑组织病理变化、神经元以及细胞凋亡情况;Western blot检测脑组织p-AMPK/AMPK、PGC1α、天冬氨酸特异性的半胱氨酸蛋白水解酶3(cysteinyl aspartate specific proteinase 3,caspase-3)和B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)蛋白表达。结果与假手术组比较,模型组脑组织神经细胞明显变性,SOD活性、神经细胞数目、Bcl-2表达明显下降(P<0.05);mNSS评分、MDA含量、凋亡神经元数目、p-AMPK/AMPK、PGC1α、caspase-3表达显著升高(P<0.05);与模型组比较,三七总皂苷组大鼠脑组织神经细胞变性现象明显改善,SOD活性、神经细胞数目、p-AMPK/AMPK、PGC1α、Bcl-2表达明显上升,mNSS评分、MDA含量、凋亡神经元数目、caspase-3表达显著降低(P<0.05);CC能明显抑制AMPK/PGC1α信号通路,加重神经细胞损伤,减弱三七总皂苷对脑挫伤后神经损伤的保护作用。结论三七总皂苷可能通过激活AMPK/PGC1α信号通路实现脑挫伤后的抗氧化、抗凋亡以及神经保护功能。
Objective To study the impacts of Panax notoginseng saponins on neuronal apoptosis in rats with brain contusion and analyze its regulatory mechanism with the AMP-activated protein kinase(AMPK)/peroxisome proliferator activated receptor gamma co-activator 1α(PGC1α)pathway.Methods Sixteen SD rats were randomly selected from ninety-eight SD rats as the sham operation group,and the remaining eighty-two rats were used to construct the traumatic brain injury(TBI)models.The 64 rats with successfully established models were divided into model group,Panax notoginseng saponins group(26.25 mg/kg),Compound C group(CC,AMPK inhibitor,of 0.25 mg/kg)and Panax notoginseng+CC(Panax notoginseng saponins of 26.25 mg/kg+CC of 0.25 mg/kg),with sixteen rats in each group.Drug treatment was carried out 24 hours after the model for 21 days.After the administration was completed,modified neurological deficit score(mNSS)scale was used to evaluate the neurological function;the levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in brain tissue of each group were detected by colorimetry;the pathological changes,neurons and apoptosis of rat brain tissue were showed by hematoxylin eosin(HE)staining,neuron-specific classⅢbeta-tubulin(Tuj1)immunofluorescence staining and terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling(TUNEL)staining;and Western blot was used to detect the expression of p-AMPK/AMPK,PGC1α,cysteinyl aspartate specific proteinase 3(caspase-3)and B-cell lymphoma-2(Bcl-2)proteins.Results Compared with the sham operation group,the nerve cells of rats in the model group were obviously degenerated,the activity of SOD,the number of nerve cells and the expression level of Bcl-2 were significantly lower;mNSS score,the content of MDA,the number of apoptotic neurons and the expression levels of p-AMPK/AMPK,PGC1α,and caspase-3 were obviously increased(P<0.05).Compared with the model group,the degeneration of nerve cells in Panax notoginseng saponins group were significantly improved,the activity of SOD,the number of nerve cells and the expression levels of p-AMPK/AMPK,PGC1αand Bcl-2 proteins were significantly higher;mNSS score,the content of MDA,the number of apoptotic neurons and the expression of caspase-3 were significantly lower(P<0.05).CC could significantly inhibit AMPK/PGC1αsignaling pathway,aggravate nerve cell damage,and weaken the protective effect of panax notoginseng saponins on nerve damage after brain contusion.Conclusion Panax notoginseng saponins may activate AMPK/PGC1αsignaling pathway to achieve anti-oxidation,anti-apoptosis,and neuroprotective functions after brain contusion.
作者
陈丹
王昕煜
江玲
唐萍
CHEN Dan;WANG Xinyu;JIANG Ling;TANG Ping(Department of Pharmacy,Chongzhou People's Hospital,Chongzhou 611230,Sichuan,China)
出处
《贵州医科大学学报》
2025年第6期840-848,共9页
Journal of Guizhou Medical University
基金
四川省医学(青年创新)科研课题项目(Q21082)。
