首页|丹酚酸B对帕金森病小鼠肠功能紊乱的保护作用及机制研究

丹酚酸B对帕金森病小鼠肠功能紊乱的保护作用及机制研究

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目的:探讨丹酚酸B(SalB)对1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的帕金森病(PD)模型小鼠的肠道功能紊乱的保护作用及机制.方法:48只C57BL/6成年雄性小鼠随机分为正常对照组(Control组)、SalB对照组(SalB组)、模型组(MPTP组)、SalB治疗组(MPTP+SalB组).腹腔注射MPTP构建PD小鼠模型,腹腔注射SalB治疗;采用爬杆实验检测小鼠行为;通过测定首粒黑便排出时间、小鼠粪便含水量评估小鼠肠道功能;免疫组化分析小鼠中脑黑质酪氨酸羟化酶(TH)阳性细胞数量、结肠Toll样受体4(TLR4)表达水平;HE染色组织切片观察小鼠结肠黏膜病理改变;免疫组化法测定小鼠黑质TH及结肠TLR4表达水平,ELISA法测定小鼠结肠组织中钙卫蛋白(CP)、TNF-α含量;Western blot检测小鼠中脑及结肠TH、结肠紧密连接蛋白(ZO-1)及TLR4/MyD88/NF-κB信号通路蛋白表达水平.结果:与正常对照组比较,模型组小鼠爬杆及转弯时间均明显增加,首粒黑便排出时间显著延长,粪便含水量明显减少;中脑TH阳性细胞数量明显减少,结肠TLR4阳性细胞数增加,中脑、结肠TH蛋白水平均显著降低,结肠黏膜病理损伤评分明显增加,结肠CP、TNF-α水平显著升高,结肠ZO-1表达明显减少;结肠TLR4、MyD88、Nuclear NF-κB p65及p-NF-κB p65蛋白表达水平均明显增加;与模型组比较,SalB治疗组小鼠爬杆及转弯时间均明显减少、首粒黑便排出时间显著缩短、粪便含水量明显增加;中脑TH阳性细胞数量明显增加,结肠TLR4阳性细胞数减少,中脑、结肠TH蛋白表达升高;结肠黏膜病理损伤评分明显降低,结肠CP、TNF-α含量显著减低,结肠ZO-1表达明显增加;结肠TLR4、MyD88、Nuclear NF-κB p65及p-NF-κB p65蛋白表达水平均明显下降.结论:SalB对MPTP诱导的PD模型小鼠具有神经及肠道双重保护作用,SalB缓解肠道炎症可能与抑制TLR4/MyD88/NF-κB炎症信号通路有关.
Effects and mechanism of Salvianolic acid B on intestinal dysfunction in mice with Parkinson's disease
Objective:To explore the effects and mechanism of Salvianolic acid B(SalB)on Parkinson's disease(PD).Methods:Forty-eight C57BL/6 male mice were randomly separated into control group(control)withoutdrugs,model group(MPTP)with intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrah-ydropyridine(MPTP),SalB control group with intraperitoneal injection of SalB,and SalB treatment group(MPTP+SalB).Construction of PD mouse model by intraperitoneal injection of MPTP,and treatment with intraperitoneal injection of SalB.Pole climbing test was applied to assess behavior differences.The time of first black stool excretion and water content of feces were measured to evaluate intestinal dysfunctions.The number of tyrosine hydroxylase(TH)positive cells in substantia nigra and the level of Toll like receptor 4(TLR4)in colon were analyzed by immunohistochemistry.The pathological changes of colonic mucosa were observed by HE staining.The levels of calprotectin(CP)and tumor necrosis factor-α(TNF-α)in colon were determined by ELISA.Western blot was used to determine the level of TH in midbrain,the protein level of TH,tight junction protein(ZO-1),and protein level of TLR4/MyD88/NF-κB signaling pathways which express in colon.Results:Com-pared with the Control group,the climbing time,T-turn time and the first black stool excretion time in MPTP group increased while the fecal water content and the number of TH positive cells in substantia nigra were decreased.Accompanied by TLR4 positive cells in colon,pathological injury score of colonic mucosa,levels of CP and TNF-α in colon increased,expression of TH in midbrain and expression of ZO-1 in colon decreased.Expressions of TLR4,MyD88,Nuclear NF-κB p65 and p-NF-κB p65 in colon increased.Com-pared with MPTP group,SalB treatment shortened the climbing time,T-turn time and the first black stool excretion time in SalB treat-ment group,increased the fecal water content and the number of TH positive cells in substantia nigra,lowered TLR4 positive cells in colon,enhanced expression of TH in midbrain and colon,reduced the pathological injury score of colonic mucosa,significantly decreased levels of CP and TNF-α in colon,enhanced expression of ZO-1 in colon,inhibited expressions of TLR4,MyD88,Nuclear NF-κB p65 and p-NF-κB p65 in colon.Conclusion:SalB can protect the nerves and intestines and alleviate the intestinal inflamma-tion of PD mice,which may be related to the inhibition of TLR4/MyD88/NF-κB signal pathway.

Salvianolic acid BParkinson's diseaseIntestinal dysfunctionGastrointestinal inflammation

南亚强、陶计委、周杰、范斐阳、蒋谷峰、朱彤、曾广红

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兰州大学第二临床医学院,兰州 730050

中国人民解放军联勤保障部队第九四〇医院,兰州 730050

丹酚酸B 帕金森病 肠功能紊乱 肠道炎症

甘肃省卫生行业科研项目中国人民解放军联勤保障部队第九四〇医院研究生导师专项

GSWSKY2018-432022yxky011

2024

中国免疫学杂志
中国免疫学会,吉林省医学期刊社

中国免疫学杂志

CSTPCD北大核心
影响因子:0.926
ISSN:1000-484X
年,卷(期):2024.40(1)
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