Effect of acupuncture on expression of endoplasmic reticulum Ca~(2+) and Caspase-12 protein in hippocampal neurons of rats with convulsive brain injury

ZHANG Hui; MA Yun; WU Xu; LIU Zhen-bang; WU Sheng-bing; ANG Wen-ping; YANG Fan

doi:10.13702/j.1000-0607.20210800

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Effect of acupuncture on expression of endoplasmic reticulum Ca~(2+) and Caspase-12 protein in hippocampal neurons of rats with convulsive brain injury

更新时间：2023-08-11

- Effect of acupuncture on expression of endoplasmic reticulum Ca~(2+) and Caspase-12 protein in hippocampal neurons of rats with convulsive brain injury
- Acupuncture Research Vol. 47, Issue 8, Pages: 673-677(2022)
- 作者机构：
  
  1. 安徽中医药大学针灸推拿学院
  2. 芜湖市中医院
  3. 中国科技大学生命科学学院
  4. 安徽中医药大学科研实验中心
- 作者简介：
- 基金信息：
- DOI：10.13702/j.1000-0607.20210800
  CLC：
- Published：2022
- 稿件说明：
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ZHANG Hui, MA Yun, WU Xu, et al. Effect of acupuncture on expression of endoplasmic reticulum Ca~(2+) and Caspase-12 protein in hippocampal neurons of rats with convulsive brain injury[J]. Acupuncture research, 2022, 47(8): 673-677.
DOI：

ZHANG Hui, MA Yun, WU Xu, et al. Effect of acupuncture on expression of endoplasmic reticulum Ca~(2+) and Caspase-12 protein in hippocampal neurons of rats with convulsive brain injury[J]. Acupuncture research, 2022, 47(8): 673-677. DOI： 10.13702/j.1000-0607.20210800.

摘要

目的:观察针刺对惊厥性脑损伤大鼠不同时点海马神经元内质网Ca

(2+)

表达、细胞凋亡和Caspase-12蛋白表达的影响，探讨针刺维持内质网钙稳态及抑制细胞凋亡级联反应的可能作用机制。方法:SD大鼠随机分为正常组、模型组、针刺组，每组36只。采用腹腔一次性注射惊厥剂量(50 mg/kg)戊四唑造成大鼠惊厥。针刺组于惊厥后即刻针刺“百会”“大椎”30 min。各组分别于惊厥发作后2、12、48 h采用TUNEL法检测海马神经元细胞凋亡情况；采用内质网Ca(2+)表达、细胞凋亡和Caspase-12蛋白表达的影响，探讨针刺维持内质网钙稳态及抑制细胞凋亡级联反应的可能作用机制。方法:SD大鼠随机分为正常组、模型组、针刺组，每组36只。采用腹腔一次性注射惊厥剂量(50 mg/kg)戊四唑造成大鼠惊厥。针刺组于惊厥后即刻针刺“百会”“大椎”30 min。各组分别于惊厥发作后2、12、48 h采用TUNEL法检测海马神经元细胞凋亡情况；采用内质网Ca

(2+)

荧光探针技术检测大鼠海马神经元内质网Ca(2+)荧光探针技术检测大鼠海马神经元内质网Ca

(2+)

表达；采用免疫组织化学法观察各组大鼠海马组织中Caspase-12蛋白表达。结果:与正常组比较，模型组2、12、48 h海马神经元凋亡细胞数明显增加(P

0.01)

海马神经元内质网Ca(2+)表达；采用免疫组织化学法观察各组大鼠海马组织中Caspase-12蛋白表达。结果:与正常组比较，模型组2、12、48 h海马神经元凋亡细胞数明显增加(P

0.01)

海马神经元内质网Ca

(2+)

表达降低(P

0.01)

Caspase-12蛋白表达升高(P

0.01);与模型组比较，针刺组在惊厥发作2、12、48 h海马神经元凋亡减轻，凋亡细胞数明显减少(P

0.05

0.01)

海马神经元内质网Ca(2+)表达降低(P

0.01)

Caspase-12蛋白表达升高(P

0.01);与模型组比较，针刺组在惊厥发作2、12、48 h海马神经元凋亡减轻，凋亡细胞数明显减少(P

0.05

0.01)

海马神经元内质网Ca

(2+)

表达升高(P

0.05

0.01)

Caspase-12蛋白表达降低(P

0.05

0.01)。结论:针刺有利于海马神经元内质网Ca(2+)表达升高(P

0.05

0.01)

Caspase-12蛋白表达降低(P

0.05

0.01)。结论:针刺有利于海马神经元内质网Ca

(2+)

表达的恢复，从而维持内质网内钙稳态，避免诱发内质网应激介导的Caspase-12凋亡级联反应，减少细胞凋亡，从而治疗惊厥性脑损伤。Objective To observe the effect of acupuncture on endoplasmic reticulum calcium

apoptosis number and Caspase-12 protein expression in hippocampal neurons of convulsive rats

so as to explore its mechanisms underlying improvement of convulsion. Methods SD rats were randomly divided into normal control

model and acupuncture groups

with 36 rats in each group. Rats in the normal control group received intraperitoneal injection(i.p.) of normal saline(2 mL)

and those of the other 2 groups received i.p. of pentylenetetrazole(50 mg/kg) for establishing convulsion model. Manual acupuncture stimulation was applied to “Baihui”(GV20) and “Dazhui”(GV14) for 30 min after modeling. The hippocampal tissues were taken at 2

12 and 48 h after modeling. The endoplasmic reticulum Ca(2+)表达的恢复，从而维持内质网内钙稳态，避免诱发内质网应激介导的Caspase-12凋亡级联反应，减少细胞凋亡，从而治疗惊厥性脑损伤。

Abstract

Objective To observe the effect of acupuncture on endoplasmic reticulum calcium

apoptosis number and Caspase-12 protein expression in hippocampal neurons of convulsive rats

so as to explore its mechanisms underlying improvement of convulsion. Methods SD rats were randomly divided into normal control

model and acupuncture groups

with 36 rats in each group. Rats in the normal control group received intraperitoneal injection(i.p.) of normal saline(2 mL)

12 and 48 h after modeling. The endoplasmic reticulum Ca

(2+)

concentration(optical density

OD) was detected by using fluorescence probe technique and laser confocal microscopy

and the number of apoptosis of hippocampal neurons at the 3 time points detected by using terminal-deoxynucleoitidyl transferase mediated nick end labeling(TUNEL) stain. The expression of Caspase-12 protein in hippocampus at 3 time points was observed by immunohistochemistry.Results In comparison with the normal group

the number of apoptotic cells of hippocampal neurons and the expression levels of Caspase-12 protein in hippocampus at 2

12 and 48 h after seizures were obviously increased(P

0.01)

and the OD value of Ca(2+) concentration(optical density

OD) was detected by using fluorescence probe technique and laser confocal microscopy

the number of apoptotic cells of hippocampal neurons and the expression levels of Caspase-12 protein in hippocampus at 2

12 and 48 h after seizures were obviously increased(P

0.01)

and the OD value of Ca

(2+)

at 3 time points significantly decreased(P

0.01) in the model group.Following acupuncture intervention

the increased levels of the number of apoptotic cells of hippocampal neurons and the expression of Caspase-12 protein in hippocampus at 3 time points and the decreased levels of OD value of Ca(2+) at 3 time points significantly decreased(P

0.01) in the model group.Following acupuncture intervention

the increased levels of the number of apoptotic cells of hippocampal neurons and the expression of Caspase-12 protein in hippocampus at 3 time points and the decreased levels of OD value of Ca

(2+)

at 3 time points were reversed in the acupuncture group(P

0.05

0.01).Conclusion Acupuncture intervention is effective in reducing the apoptosis of hippocampal neurons in convulsion rats

which may be related to its functions in down-regulating Caspase-12 expression and promoting influx of Ca(2+) at 3 time points were reversed in the acupuncture group(P

0.05

0.01).Conclusion Acupuncture intervention is effective in reducing the apoptosis of hippocampal neurons in convulsion rats

which may be related to its functions in down-regulating Caspase-12 expression and promoting influx of Ca

(2+)

in the hippocampal neurons.

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