云南哈尼族验方“魔呐奇”抑制缺血半影区神经元自噬流诱导卒中后神经保护研究  

Yunnan hani nationality prescription“Monaqi”inhibits the neuronal autophagic flow in ischemic penumbra area to induce post-stroke neuroprotection

作　　者：张雪玉 邓仪昊[2] 郭涛[2] 臧瑞 武煜明 蔡恩丽 ZHANG Xueyu;DENG Yi Hao;GUO Tao;ZANG Rui;WU Yuming;CAI Enli(Yunnan University of Traditional Chinese Medicine,Kunming 650000,Yunnan,China;Kunming University of Science and Technology,Kunming 650000,Yunnan,China)

机构地区：[1]云南中医药大学,云南昆明650000 [2]昆明理工大学,云南昆明650000

出　　处：《现代中西医结合杂志》2021年第16期1711-1716,共6页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：国家自然科学基金项目(81960418,81860411);云南中医学院、云南省科技厅联合专项(2017FF116,2017FF117);云南省应用基础研究计划项目(2019FB098)。

摘　　要：目的观察云南哈尼族验方“魔呐奇”对大鼠缺血性脑卒中后半影区神经元自噬流的影响。方法将54只SD大鼠随机分为假手术组、模型组和魔呐奇组各18只,模型组和魔呐奇组参照线栓法制备大脑中动脉梗死模型。造模24 h后,魔呐奇组给予“魔呐奇”洒剂腹腔注射,模型组腹腔注射等量生理盐水。灌胃14 d后,评估神经功能缺损评分,TTC染色测定大脑相对梗死体积;Western blot法检测缺血半影区自噬流通路相关蛋白Beclin-1、LC3、P62(Insouble,Souble)、Ubiquitin、CathepsinB、LAMP1表达水平;免疫荧光双标法对皮质缺血半影区LC3阳性神经元蛋白表达进行定位,计算阳性细胞占比。结果模型组大鼠mNSS评分为(5.16±0.98)分,梗死脑组织占比为(23.87±3.78)%,魔呐奇组分别为(2.67±0.52)分和(18.35±1.75)%,魔呐奇组均明显低于模型组(P均<0.05)。模型组大鼠缺血半影区Beclin-1、LC3Ⅱ/LC3Ⅰ、P62(Insouble)、Ubiquitin、CathepsinB、LAMP1蛋白表达水平均明显高于假手术组(P均<0.05),P62(Souble)蛋白表达水平明显低于假手术组(P<0.05);魔呐奇组大鼠缺血半影区Beclin-1、LC3Ⅱ/LC3Ⅰ、P62(Insouble)、Ubiquitin、CathepsinB、LAMP1蛋白表达水平均明显低于模型组(P均<0.05),P62(Souble)蛋白表达水平与模型组比较差异无统计学意义(P>0.05);模型组皮质缺血半影区LC3阳性表达占比明显高于假手术组(P<0.05),魔呐奇组明显低于模型组(P<0.05)。结论“魔呐奇”可抑制皮质缺血半影区自噬流,保持自噬与溶酶体清除动态平衡,提高自噬流通畅性,发挥卒中后神经保护作用。Objective It is to observe the effect of the prescription“Monaqi”of Hani nationality in Yunnan on the autophagic flux of neurons in the penumbra area after ischemic stroke in rats.Methods 54 SD rats were randomly divided into sham operation group,model group and Monaqi group,with 18 rats in each group.Model group and Monaqi group were used to prepare middle cerebral artery infarction models with reference to the suture method.24 hours after the model was established,the Monaqi group was given the“Monaqi”decoction by gavage,and the sham operation group and the model group were given the same amount of normal saline.After intragastric administration for 14 days,the neurological deficit score was evaluated,the relative infarct volume of the brain was detected by TTC staining;the expression levels of the autophagy pathway related proteins Beclin-1,LC3,P62(Insouble,Souble),Ubiquitin,CathepsinB,LAMP1 in the penumbra area were detected by Western blot method;The immunofluorescence double-labeling method was used to locate the LC3 positive cells in the cortical ischemic penumbra and calculate the proportion of positive cells.Results The mNSS score of rats in the model group was(5.16±0.98)points,the proportion of infarcted brain tissue was(23.87±3.78)%,and the mNSS scores and proportion of infarcted brain tissue in the Monaqi group were(2.67±0.52)points and(18.35±1.75)%,respectively,the mNSS score and the proportion of infarcted brain tissue in the Monaqi group were significantly lower than those in the model group(all P<0.05).The expression levels of Beclin-1,LC3II/LC3I,P62(Insouble),Ubiquitin,CathepsinB,LAMP1 in the ischemic penumbra of the model group were significantly higher than those in the sham operation group(all P<0.05),and the expression level of P62(Souble)protein was significantly lower than that of the sham operation group(P<0.05);the protein expression levels of Beclin-1,LC3Ⅱ/LC3Ⅰ,P62(Insouble),Ubiquitin,CathepsinB,and LAMP1 in the ischemic penumbra of rats in the Monaqi group were significa

关 键 词：魔呐奇 自噬流 神经保护 缺血性脑卒中 溶酶体功能 

分 类 号：R-332[医药卫生]