白芍总苷对帕金森综合征大鼠的神经保护作用及机制研究  

Neuroprotective role and mechanism of total glucosides of paeony on Parkinson syndrome rats

作　　者：鲁晓玲 孙勤国 黄智辉 丁晓明 Lu Xiaoling;Sun Qinguo;Huang Zhihui;Ding Xiaoming(Department of Traditional Chinese Medicine,Wuhan Third Hospital,Wuhan 430060,Hubei Province,China)

机构地区：[1]武汉市第三医院中医科,430060

出　　处：《中华老年心脑血管病杂志》2025年第2期223-228,共6页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：武汉市中医药科研项目(WZ21A05)。

摘　　要：目的探讨白芍总苷对帕金森综合征(Parkinson syndrome,PS)大鼠的神经保护及对Ste20相关的富含脯氨酸-丙氨酸激酶/阳离子-氯离子共转运体家族中的钠钾氯共转运体亚型1(Ste20-like proline-/alanine-rich kinase/Na^(+)-K^(+)-Cl-cotransporter 1,SPAK/NKCC1)信号通路的影响。方法选取60只7周龄雄性SD大鼠构建PS模型,全部造模成功,按造模和给药干预的不同随机分为模型组、白芍总苷低剂量组、白芍总苷高剂量组、白芍总苷高剂量+阴性对照组、白芍总苷高剂量+丝氨酸/苏氨酸蛋白激酶3(WNK lysine deficient protein kinase 3,WNK3)过表达组,每组12只,另取12只正常健康大鼠作为对照组。白芍总苷低剂量组、白芍总苷高剂量组分别给予50、200 mg/kg白芍总苷灌胃;白芍总苷高剂量+阴性对照组与白芍总苷高剂量+WNK3过表达组灌胃200 mg/kg白芍总苷并分别尾静脉注射阴性对照质粒、WNK3质粒;对照组给予等量生理盐水;均1次/d,连续给药7 d。酶联免疫吸附测定检测血清白细胞介素(interleukin,IL)-6、IL-1β、丙二醛(malondialdehyde,MDA)、超氧化物歧化酶(superoxide dismutase,SOD)水平;HE染色检测脑组织黑质区病理形态;TUNEL染色检测神经元细胞凋亡;免疫组织化学检测α-突触核蛋白(α-Synuclein,α-Syn)表达;Western blot检测Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)及SPAK/NKCC1信号通路相关蛋白[WNK3、磷酸化SPAK(p-SPAK)、SPAK、磷酸化NKCC1(p-NKCC1)、NKCC1]表达。结果与模型组比较,白芍总苷低、高剂量组黑质区神经元病理损伤减轻,IL-6、IL-1β、MDA水平及神经元凋亡率、Bax、α-syn、WNK3、p-SPAK/SPAK、p-NKCC1/NKCC1表达降低,SOD水平及Bcl-2表达升高[(88.39±8.96)U/mg,(119.57±12.01)U/mg vs(60.28±6.14)U/mg,P<0.05;0.57±0.06,0.82±0.09 vs 0.38±0.04,P<0.05];与白芍总苷高剂量+阴性对照组比较,白芍总苷高剂量+WNK3过表达组黑质区神经元病理损伤加重Objective To investigate the neuroprotective effect of TGP on PS rats and determine the impact on the Ste20-like proline/alanine-rich kinase/Na^(+)-K^(+)-Cl cotransport(SPAK/NKCC1)signaling pathway.Methods After PS model was successfully established in 60male SD rats(7 weeks old),they were randomly divided into model group,low-and high-dose TGP groups,highdose TGP+negative control group,and high-dose TGP+WNK3overexpression group,with 12 rats in each group.Another 12healthy rats served as the control group.After modeling,50or 200 mg/kg TGP was given to the rats of corresponding groups intragastrically,the overexpression plasmids of WNK3were given to the rats from the high-dose TGP+WNK3overexpression group through tail vein injection,and same volume of normal saline was given to the control group.All of these agents were administrated once per day for 7consecutive days.ELISA was applied to detect serum levels of IL-6,IL-1β,MDA and SOD.HE staining was applied to detect the pathological morphology of the substantia nigra region in brain tissue.TUNEL staining was used to observe neuronal apoptosis.Immunohistochemistry was conducted to measure the expression ofα-synuclein(α-syn),and Western blotting for the expression of Bax,Bcl-2and SPAK/NKCC1signaling pathway related proteins(WNK3,p-SPAK,SPAK,p-NKCC1and NKCC1).Results Compared with the model group,the pathological damage of neurons in substantia nigra was reduced in lowand high dose TGP groups,reduced contents of IL-6,IL-1βand MDA,lower neuronal apoptotic rate,and declined expression of Bax,α-syn,WNK3,p-SPAK/SPAK,and p-NKCC1/NKCC1,but raised SOD content and Bcl-2expression level(88.39±8.96U/mg,119.57±12.01U/mg vs 60.28±6.14U/mg,P<0.05;0.57±0.06,0.82±0.09 vs 0.38±0.04,P<0.05).The intervention with WNK3overexpression resulted in more severe pathological damage to neurons in the substantia nigra,increased contents of IL-6,IL-1βand MDA,higher neuronal apoptotic rate,enhanced expression of Bax,α-syn,WNK3,p-SPAK/SPAK,and p-NKCC1/NKCC1,and reduced SOD content a

关 键 词：大鼠 帕金森障碍 神经保护 研究 

分 类 号：R28[医药卫生—中药学]