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1.南京中医药大学附属医院/江苏省中医院,南京 210029
2.南京中医药大学针药结合教育部重点实验室,江苏 南京 210023
3.安徽中医药大学第一附属医院,合肥 230031
4.深圳市宝安区中医院
5.安徽中医药大学
△王萍,南京中医药大学博士研究生。E-mail:wping1223@163.com
✉刘兰英,主任医师。E-mail:bluelany@163.com
收稿:2024-01-15,
网络首发:2024-08-28,
纸质出版:2025-01-12
移动端阅览
王萍, 杨骏, 孔钰, 等. 基于海马Mst1/NF-κB p65通路探讨化瘀通络灸对血管性痴呆大鼠学习记忆能力的影响[J]. 中国针灸, 2025,45(1):53-60.
WANG Ping, YANG Jun, KONG Yu, et al. Effect of
王萍, 杨骏, 孔钰, 等. 基于海马Mst1/NF-κB p65通路探讨化瘀通络灸对血管性痴呆大鼠学习记忆能力的影响[J]. 中国针灸, 2025,45(1):53-60. DOI: 10.13703/j.0255-2930.20240115-k0001.
WANG Ping, YANG Jun, KONG Yu, et al. Effect of
目的:
2
观察化瘀通络灸对血管性痴呆(VD)大鼠学习记忆能力及海马炎性反应相关通路哺乳动物不育系20样激酶1(Mst1)/核转录因子κB(NF-κB)p65的影响。
方法:
2
将60只SPF级雄性Wistar大鼠随机分为假手术组(12只)和造模组(48只),造模组大鼠采用改良双侧颈总动脉永久结扎方法建立VD模型,将造模成功的36只大鼠随机分为模型组、艾灸组和西药组,每组12只。艾灸组大鼠予化瘀通络灸,穴取“大椎”“百会”“神庭”,每次20 min,每日1次,7 d为一疗程,疗程间休息1 d,共干预3个疗程。西药组予0.72 mg/kg吡拉西坦灌胃,每天2次,疗程与艾灸组相同。应用Morris水迷宫实验检测各组大鼠学习记忆能力,HE染色法观察大鼠海马CA1区形态,实时荧光定量PCR法检测大鼠海马Mst1、M1型小胶质细胞标志物CD86、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)mRNA表达,ELISA法检测大鼠海马IL-6、TNF-α含量,Western blot法检测大鼠海马Mst1、NF-κB p65蛋白表达。
结果:
2
与假手术组比较,模型组大鼠逃避潜伏期延长(
P
<
0.05);与模型组比较,艾灸组和西药组大鼠逃避潜伏期均缩短(
P
<
0.05)。模型组大鼠海马CA1区细胞排列无序,细胞塌陷,细胞核不规则;与模型组比较,艾灸组和西药组大鼠海马CA1区细胞排列较规整,损伤程度好转。与假手术组比较,模型组大鼠海马Mst1、CD86、IL-6、TNF-α mRNA及Mst1、NF-κB p65蛋白表达升高(
P
<
0.05);与模型组比较,艾灸组和西药组大鼠海马Mst1、CD86、IL-6、TNF-α mRNA及Mst1、NF-κB p65蛋白表达降低(
P
<
0.05)。与假手术组比较,模型组大鼠海马IL-6、TNF-α含量升高(
P
<
0.05);与模型组比较,艾灸组和西药组大鼠海马IL-6、TNF-α含量降低(
P
<
0.05)。
结论:
2
化瘀通络灸可改善VD大鼠学习记忆能力,作用机制可能为通过Mst1/NF-κB p65通路调控小胶质细胞激活,减少促炎因子IL-6、TNF-α的释放,从而缓解炎性因子对VD大鼠海马的损害。
Objective
2
To obser
ve the effects of
Huayu Tongluo
(transforming stasis and unblocking collaterals) moxibustion on learning-memory ability and hippocampal mammalian sterile 20-like kinase 1 (Mst1)/nuclear factor κB (NF-κB) p65 pathway related to inflammatory response in rats with vascular dementia (VD).
Methods
2
A total of 60 male Wistar rats of SPF grade were randomly divided into a sham operation group (12 rats) and a modeling group (48 rats). VD model was established by the method of modified bilateral common carotid artery permanent ligation in the modeling group. Thirty-six rats with successful modeling were randomly divided into a model group
a moxibustion group and a western medication group
with 12 rats in each group.
Huayu Tongluo
moxibustion was applied at "Dazhui" (GV14)
"Baihui" (GV20) and "Shenting" (GV24) in the moxibustion group
20 min each time
once a day
7 day-intervention was as one course
and 1 day-interval was taken between two courses
for a total of 3 courses. In the western medication group
piracetam was given 0.72 mg/kg by intragastric administration
twice a day
the course of intervention was same as that of the moxibustion group. The learning-memory ability was detected by Morris water maze test; the morphology of hippocampal CA1 region was observed by HE staining; the mRNA expression of Mst1
M1 microglia markers CD86
interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) was detected by real-time PCR; the levels of IL-6 and TNF-α in hippocampus were detected by ELISA; and the protein expression of Mst1 and NF-κB p65 in hippocampus was detected by Western blot in rats of each group.
Results
2
Compared with the sham operation group
the escape latency was prolonged in the model group (
P
<
0.05); compared with the model group
the escape latency was shortened in the moxibustion group and the western medication group (
P
<
0.05). The cells in the CA1 region of hippocampu
s were disordered
cell collapse and irregular nuclei could be observed in the model group; compared with the model group
the cell arrangement in the CA1 region of hippocampus was more regular
and the damage was improved in the moxibustion group and the western medication group. Compared with the sham operation group
the mRNA expression of Mst1
CD86
IL-6 and TNF-α
as well as the protein expression of Mst1
NF-κB p65 in hippocampus were increased in the model group (
P
<
0.05). Compared with the model group
the mRNA expression of Mst1
CD86
IL-6 and TNF-α
as well as the protein expression of Mst1
NF-κB p65 in hippocampus were decreased in the moxibustion group and the western medication group (
P
<
0.05). Compared with the sham operation group
the levels of IL-6 and TNF-α in hippocampus were increased in the model group (
P
<
0.05). Compared with the model group
the levels of IL-6 and TNF-α in hippocampus were decreased in the moxibustion group and the western medication group (
P
<
0.05).
Conclusion
2
Huayu Tongluo
moxibustion can improve the learning-memory ability of VD rats
the mechanism may be related to regulating the activation of microglia through Mst1/NF-κB p65 pathway
reducing the release of pro-inflammatory factors i.e. IL-6 and TNF-α
so as to alleviating the damage of inflammatory factors in the hippocampus of VD rats.
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