Jia Lan-Yu, Du Yuan-Hao
Tianjin Academy of Traditional Chinese Medicine Affiliated Hospital, Tianjin 300120, China.
The First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300193.
Zhen Ci Yan Jiu. 2021 Nov 25;46(11):914-20. doi: 10.13702/j.1000-0607.201141.
To observe the effect of electroacupuncture (EA) of "Shuigou" (GV26) on the activities of sonic hedgehog(Shh) signaling molecules (Shh, Ptch, Smo，Gli and Gli2) in ischemic brain tissues in rats with cerebral ischemia (CI), so as to explore its mechanisms underlying improvement of CI.
Male Wistar rats were randomly divided into blank control group(=10), sham operation group (=90), model group(=90) and EA group (=90). The CI model was established by occlusion of the right middle cerebral artery (MCAO). According to the postoperative time points of MCAO, the later three groups were further divided into 1, 3, 6, 9, 12 and 24 h, and 3, 7 and 12 d subgroups, with 10 rats in each subgroup. EA (15 Hz, 2 mA) was applied to GV26 for 20 min. The 1 h to 24 h subgroups were treated immediately after modeling, the 3-12 d subgroups treated one time a day. The neurological severity score (NSS, 0 to 18 points) was used to evaluate the rats' neurological function, and TTC staining was employed to assess the cerebral ischemic volume (percentage of cerebral infarct volume, CIV). Western blot was employed to detect the expression of Shh, Ptch, Smo, Gli1 and Gli2 proteins in the ischemic cerebral tissue.
Compared with the sham operation group, the NSS scores of the model group increased at all time points (<0.01). The percentages of CIV of the model group from 3 h to 12 d were obviously higher than those of the sham operation group (<0.01). The NSS scores at 3, 7 and 12 d and the percentages of CIV at 1, 3, 7 and 12 d after MCAO were significant lower in the EA group than in the model group (<0.05). The protein expression levels of Shh from 12 h to 12 d (i.e. 12 h, 24 h, 3, 7 and 12 d), Ptch from 6 h to 12 d, Smo from 9 h to 12 d, Gli1 at 9 h, 12 h, and from 3 d to 12 d, Gli2 at 6, 9 and 12 h, and 3 d were significantly higher in the model group than in the sham operation group (<0.05, <0.01), while those of Shh at 3, 7 and 12 d, Ptch from 24 h to 7 d, Smo from 12 h to 7 d, Gli1 from 24 h to 7 d, Gli2 at 12 h, 3 and 7 d were significantly higher in the EA group than in the model group (<0.05, <0.01). No statistical significances were found between the sham operation and the blank control groups in all the indexes mentioned above (>0.05).
EA of GV26 can improve neurological function and reduce infarct volume in MCAO rats, which may be related to its function in up-regulating the activities of Shh signaling pathway in the ischemic cerebral tissues.
观察电针“水沟”（GV26）对脑缺血（CI）大鼠缺血脑组织中声波刺猬因子（Shh）信号分子（Shh、Ptch、Smo、Gli和Gli2）活性的影响，以探讨其改善CI的作用机制。
将雄性Wistar大鼠随机分为空白对照组（n = 10）、假手术组（n = 90）、模型组（n = 90）和电针组（n = 90）。采用右侧大脑中动脉闭塞法（MCAO）制备CI模型。根据MCAO术后时间点，后三组进一步分为1、3、6、9、12和24 h，以及3、7和12 d亚组，每个亚组10只大鼠。将电针（15 Hz，2 mA）施加于GV26 20 min。1 h至24 h亚组在造模后立即治疗，3 - 12 d亚组每天治疗1次。采用神经功能缺损评分（NSS，0至18分）评估大鼠神经功能，采用TTC染色评估脑缺血体积（脑梗死体积百分比，CIV）。采用蛋白质免疫印迹法检测缺血脑组织中Shh、Ptch、Smo、Gli1和Gli2蛋白的表达。
与假手术组相比，模型组各时间点NSS评分均升高（P < 0.01）。模型组3 h至12 d的CIV百分比明显高于假手术组（P < 0.01）。MCAO术后3、7和12 d时，电针组的NSS评分及1、3、7和12 d时的CIV百分比均显著低于模型组（P < 0.05）。模型组12 h至12 d（即12 h、24 h、3、7和12 d）Shh、6 h至12 d Ptch、9 h至12 d Smo、9 h、12 h以及3 d至12 d Gli1、6、9和12 h以及3 d Gli2的蛋白表达水平显著高于假手术组（P < 0.05，P < 0.01），而电针组3、7和12 d时Shh、24 h至7 d Ptch、12 h至7 d Smo、24 h至7 d Gli1、12 h、3和7 d Gli2的蛋白表达水平显著高于模型组（P < 0.05，P < 0.01）。上述所有指标在假手术组和空白对照组之间均无统计学差异（P > 0.05）。
电针GV26可改善MCAO大鼠的神经功能，减小梗死体积，这可能与其上调缺血脑组织中Shh信号通路活性的作用有关。
