【化瘀通络灸通过调节Sonic Hedgehog信号通路提高血管性痴呆大鼠学习记忆能力,促进髓鞘再生】。

Yu Kong, Jun Yang, Yin-Qiu Fan, Wei-Shun Wang, Ya-Ting Zhang
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摘要

目的:观察化瘀通络灸对血管性痴呆(VD)大鼠胼胝体髓鞘再生及Shh信号通路的影响,探讨其改善VD的机制。方法:将雄性Wistar大鼠随机分为假手术组、模型组、给药组和艾灸组,每组12只。采用双侧颈总动脉闭塞法建立VD模型。灸“神庭”(GV24)、“百会”(GV20)、“大椎”(GV14)各20分钟,每日1次,7 d为1个疗程,共3个疗程,每2个疗程中间休息1天。给药组大鼠给予氯马司汀溶液10 mg/kg灌胃,每日1次,疗程与艾灸组相同。Morris水迷宫实验(逃避潜伏期)评估大鼠学习记忆能力。采用龙格量表评估神经功能缺损。采用实时荧光定量PCR和Western blot分别检测脑胼胝体中Shh和Gli1 mRNA和蛋白的表达。透射电镜观察大鼠髓鞘和髓鞘轴突超微结构。结果:与假手术组比较,脑缺血大鼠神经系统评分和逃避潜伏期均显著升高、延长(ppppp)。结论:化瘀通络灸可通过调节Shh信号通路中Shh和Gli1的表达,促进脑缺血后少突胶质前细胞的分化成熟,从而促进脑白质髓鞘的再生,可能有助于提高学习记忆能力。
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["Huayu Tongluo" moxibustion improves learning-memory ability and promotes myelin regeneration by regulating Sonic Hedgehog signaling pathway in vascular dementia rats].

Objective: To investigate the effect of "Huayu Tongluo" (resolving blood stagnation to dredge meridian-collaterals) moxibustion on remyelination and Sonic Hedgehog (Shh) signaling pathway in the corpus callosum of vascular dementia (VD) rats, so as to explore its mechanisms underlying improvement of VD.

Methods: Male Wistar rats were randomized into sham-operation, model, medication and moxibustion groups, with 12 rats in each group.The VD model was established by bilateral common carotid artery occlusion. Moxibustion was applied to "Shenting"(GV24), "Baihui"(GV20) and "Dazhui"(GV14) for 20 min once a day, 7 d as a treatment course, for 3 courses, with one day's rest between every two courses. Rats of the medication group were treated by gavage of 10 mg/kg of chloromastine solution once a day, and the course of treatment was the same as that of the moxibustion group. The rat's learning-memory ability was assessed by Morris water maze test (escape latency). The neurological deficits were evaluated by using Longa's scale.The mRNA and protein expressions of Shh and Gli1 in the corpus callosum were measured by quantitative real-time fluorescence PCR and Western blot, separately. The ultrastructure of the myelin sheath and myelinated axons was observed under transmission electron microscopy (TCM).

Results: Compared with the sham-operation group, the neurologic score and escape latency were significantly increased and prolonged (P<0.01), and the mRNA and protein expression levels of Shh and Gli1 and the number of myelinated axons were obviously decreased in the model group (P<0.01). In comparison with the model group, the escape latency was apparently shortened (P<0.05), while the mRNA and protein expression levels of Shh and Gli1 as well as the number of myelinated axons were strikingly increased in both moxibustion and medication groups (P<0.01). Results of TCM showed that in the model group, the arrangement of myelin coil structures was sparse and fuzzy, and some structures were bulged and disbanded. The oligodendrocytes were irregular, and the number of myelin sheath was rare. These situations were relatively milder in both moxibustion and medication groups.

Conclusion: "Huayu Tongluo" moxibustion can promote the differentiation and maturation of oligodendrocyte precursor cells after cerebral ischemia by regulating the expressions of Shh and Gli1 in Shh signaling pathway, thus promoting the regeneration of cerebral white matter myelin sheaths in VD rats, which may contribute to improving learning-memory ability.

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