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1. 江西中医药大学附属医院
2. 上海市第八人民医院中医科
3. 南昌医学院中医学院
4. 上海市顺昌医院中医科
纸质出版日期:2022
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闵友江, 洪冬英, 姚海华, 等. 艾灸对脑缺血再灌注损伤大鼠脑梗死区Nogo/神经营养素受体信号通路的影响[J]. 针刺研究, 2022,47(5):401-408.
MIN You-jiang, HONG Dong-ying, YAO Hai-hua, et al. Effect of moxibustion on Nogo/neurotrophin receptor signaling pathway in rats with cerebral ischemia/reperfusion injury[J]. Acupuncture research, 2022, 47(5): 401-408.
闵友江, 洪冬英, 姚海华, 等. 艾灸对脑缺血再灌注损伤大鼠脑梗死区Nogo/神经营养素受体信号通路的影响[J]. 针刺研究, 2022,47(5):401-408. DOI: 10.13702/j.1000-0607.20210592.
MIN You-jiang, HONG Dong-ying, YAO Hai-hua, et al. Effect of moxibustion on Nogo/neurotrophin receptor signaling pathway in rats with cerebral ischemia/reperfusion injury[J]. Acupuncture research, 2022, 47(5): 401-408. DOI: 10.13702/j.1000-0607.20210592.
目的:观察艾灸治疗对脑缺血再灌注损伤(CI/RI)大鼠脑梗死区Nogo-A、Nogo-A受体(NgR)、协同受体p75神经营养素受体(p75NTR)与亮氨酸富集重复片段和Ig域包含子(Lingo-1)表达的影响,探讨艾条悬灸治疗CI/RI的机制。方法:雄性SD大鼠按随机数字表法分为假手术组、模型组、阻断剂组和艾灸组,假手术组16只,其余每组17只。采用线栓法复制左侧大脑中动脉栓塞模型。造模后,艾灸组用艾条悬灸“百会”和右侧“曲池”“足三里”20 min
每日1次,7次为1个疗程,疗程中间休息2 d
共治疗2个疗程;阻断剂组硬膜下推注Nogo分子胞外段氨基端前40个残基多肽(NEP1-40)。干预前后对各组大鼠进行神经功能评分,干预后以Feeney走横木实验(BWT)评分对其精细运动功能恢复情况进行评估;用TTC染色法评估大鼠脑梗死体积;实时荧光定量PCR法检测梗死区脑组织Nogo-A、NgR、p75NTR、Lingo-1 mRNA表达;Western blot法检测梗死区脑组织Nogo-A、NgR、p75NTR、Lingo-1蛋白表达;免疫荧光双标法检测梗死侧脑组织NgR、Lingo-1的阳性表达。结果:与假手术组比较,模型组大鼠神经功能评分、脑梗死体积百分比均明显升高(P<0.01)
BWT评分明显下降(P<0.01)
梗死区脑组织Nogo-A、NgR、p75NTR、Lingo-1 mRNA及蛋白表达均明显升高(P<0.01)
NgR和Lingo-1共表达增加(P<0.01)。与模型组比较,除阻断剂组Nogo-A蛋白表达外,两干预组大鼠上述其余指标都明显逆转(P<0.01)。与阻断剂组比较,艾灸组大鼠脑梗死体积百分比、梗死区脑组织Nogo-A、p75NTR mRNA表达,Nogo-A、NgR、Lingo-1、p75NTR蛋白表达及NgR和Lingo-1共表达均降低(P<0.01
P<0.05)。结论:艾灸能产生类似于阻断剂NEP1-40的作用,抑制脑梗死后Nogo/神经营养素受体信号通路相关分子的表达,这可能是艾灸治疗脑梗死的机制之一。
Objective To observe the effect of moxibustion treatment on the expression of Nogo-A
Nogo receptor(NgR)
neurotrophin receptor p75(p75 NTR) and leucine rich repeat and Ig domain containing 1(Lingo-1) in brain tissue of rats with cerebral ischemia/reperfusion injury(CI/RI)
so as to analyze its mechanism underlying improvement of CI/RI. Methods Male SD rats were randomly divided into sham operation group(16 rats)
model group(17 rats)
NEP1-40(extracellular peptide residues 1-40
a blocker targeting NgR) group(model+blocker
17 rats) and moxibustion group(model+moxibustion
17 rats). The CI/RI model was established by occlusion of the left middle cerebral artery(MCAO). Moxibustion was applied to “Baihui”(GV20)
right “Quchi”(LI11) and “Zusanli”(ST36) for 20 min
once a day for 14 days
with 2 days' rest after the top 7 days' intervention. For rats of the NEP1-40 group
30 μL PBS containing 18 μg NEP 1-40 was injected into the epidural inferior vena(L5-S1) via a polyvinyl chloride conduit. The neurological deficit state in each group was evaluated by Longa's 5-point scale and Feeney's 7-point scale of beam walking test(BWT). The cerebral infarct volume was assessed by 2
3
5-triphenyltetrazole chloride staining. The brain tissue between the central anterior and posterior sulcus was taken for observing the expression of NgR and Lingo-1 by fluorescence double-label method
and for determining the expression levels of Nogo-A
NgR
p75 NTR and Lingo-1 mRNAs and proteins by real-time quantitative PCR and Western blot
respectively. Results After modeling
the Longa's score
infarct volu-me percent
expression levels of Nogo-A
NgR
Lingo-1 and p75 NTR mRNAs and proteins were significantly increased(P<0.01) and BWT score was obviously decreased(P<0.01) in the model group relevant to the sham operation group. In comparison with the model group
the increase of Longa's score
infarct volume percentage
expression levels of Nogo-A
NgR
Lingo-1 and p75 NTR mRNAs and proteins and decrease of BWT score in NEP1-40 and moxibustion groups were reversed(P<0.01) except Nogo-A protein in the NEP1-40 group. The effect of moxibustion was significantly superior to that of blocker NEP1-40 in redu-cing the infarct volume percentage
and down-regulating the expression of Nogo-A mRNA and protein
p75 NTR mRNA and protein
NgR and Lingo-1 proteins(P<0.01
P<0.05). Conclusion Moxibustion
similar to blocker NEP1-40 of NgR
can improve neurological dysfunction in CI/RI rats
which may be related to its functions in reducing cerebral infarction and down-regulating the activity of Nogo/neurotrophin receptor signaling pathway.
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