Effect of Shuyu Capsule on Cav1.2-mediated CaM/CaMK Ⅱ Signaling Pathway in Rats with PMDD Liver-Qi Stagnation Syndrome Model

LI Yi-jie; WANG Mei-yan; XUE Ling; SONG Chun-hong

doi:10.13422/j.cnki.syfjx.20181034

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Effect of Shuyu Capsule on Cav1.2-mediated CaM/CaMK Ⅱ Signaling Pathway in Rats with PMDD Liver-Qi Stagnation Syndrome Model

更新时间：2024-01-04

- Effect of Shuyu Capsule on Cav1.2-mediated CaM/CaMK Ⅱ Signaling Pathway in Rats with PMDD Liver-Qi Stagnation Syndrome Model
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 10, Pages: 130-136(2018)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.20181034
  CLC： R-332;R285.5
- Published：2018
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LI Yi-jie, WANG Mei-yan, XUE Ling, et al. Effect of Shuyu Capsule on Cav1.2-mediated CaM/CaMK Ⅱ Signaling Pathway in Rats with PMDD Liver-Qi Stagnation Syndrome Model[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(10): 130-136.
DOI：

LI Yi-jie, WANG Mei-yan, XUE Ling, et al. Effect of Shuyu Capsule on Cav1.2-mediated CaM/CaMK Ⅱ Signaling Pathway in Rats with PMDD Liver-Qi Stagnation Syndrome Model[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(10): 130-136. DOI： 10.13422/j.cnki.syfjx.20181034.

摘要

目的：观察舒郁胶囊对强迫游泳应激诱导的经前烦躁障碍（premenstrual dysphoric disorder，PMDD）肝气郁证大鼠的影响，探讨L型钙通道（L-type calcium channel，LTCC）Cav1.2亚型介导的钙调蛋白（CaM）/CaMKⅡ信号通路在PMDD肝气郁证发生中的机制。方法：动情周期规律的大鼠进行3次强迫游泳实验，筛选出48只雌性Wistar大鼠进入实验，两次非接受期和接受期悬浮不动时间差值绝对值平均值最小的12只为正常组，其余36只分为模型组、舒郁胶囊组（0.408 g·kg-1·d-1），氟西汀组（2.7 mg·kg-1·d-1）。给药组连续给药2个动情周期，模型组和正常组给以等量纯水。采用旷场实验、强迫游泳悬浮不动时间及悬浮不动次数评价模型和药物干预效果。免疫荧光、实时荧光定量聚合酶链式反应（Real-time PCR）和蛋白质免疫印迹（Western blot）检测各组大鼠海马脑区中L型钙通道α1C亚型（CACNA1C），CaM依赖性蛋白激酶Ⅱ（CaMKⅡ）的分布及表达。结果：CACNA1C蛋白主要分布在细胞膜上，CaMKⅡ蛋白主要分布在的细胞质内；与正常组比较，模型组大鼠体质量、旷场总路程显著下降（P<0.05，P<0.01），强迫游泳悬浮不动时间、悬浮不动次数显著升高（P<0.05，P<0.01），海马脑区细胞排列散乱，CACNA1C和CaMKⅡ表达升高（P<0.05，P<0.01）；与模型组比较，氟西汀和舒郁胶囊组大鼠体质量、旷场总路程明显升高（P<0.05，P<0.01），强迫游泳悬浮不动时间、悬浮不动次数明显降低（P<0.05，P<0.01），海马脑区细胞排列整齐，CACNA1C和CaMKⅡ表达明显降低（P<0.05，P<0.01）。结论：利用强迫游泳可成功诱导出PMDD肝气郁证大鼠模型。舒郁胶囊可能通过Cav1.2介导的CaM/CaMKⅡ信号通路发挥治疗作用，显著改善PMDD肝气郁证大鼠的行为学变化。

Abstract

Objective: To observe the effect of Shuyu capsule on premenstrual dysphoric disorder (PMDD) type liver-qi depression rats induced by forced swimming test

in order to investigate the mechanism of L-type calcium channel (LTCC) Cav1.2 subtype-mediated Calmodulin (CaM)/CaMK Ⅱ signaling pathway in the pathogenesis of PMDD stagnation of liver-Qi. Method: Totally 48 female Wistar rats with regular estrous cycle were selected by three forced swimming. The mean absolute value of difference value between non-acceptance period and suspension time was calculated. The 12 rats with the lowest mean value was included in the normal group

and the remaining 36 rats were divided into model group

Shuyu capsule group (0.408 g·kg-1·d-1) and fluoxetine administration group (2.7 mg·kg-1·d-1). The drug group was given for continuously two estrous cycles

and the model group and the normal group were given the same amount of pure water. The open field test

immobility duration and immobility frequency were used to evaluate the model and drug intervention effect. The distribution and expressions of L-type calcium Channel α1C (CACNA1C) and CaMK Ⅱ genes in hippocampus of rats were detected by immunofluorescence

real-time fluorescence quantitative PCR and Western blot. Result: CACNA1C protein was mainly distributed in the cell membrane

CaMK Ⅱ protein was mainly distributed in the cytoplasm. Compared with the normal group

the body weight and total distance of open field text of model group were significantly decreased (P<0.05

P<0.01)

immobility duration and immobility frequency of model group were significantly increased (P<0.05

P<0.01)

hippocampus cells was scattered

CACNA1C and CaMK Ⅱ expressions were increased (P<0.05

P<0.01) in model group. Compared with the model group

the body weight and total distance of open field text of model group were significantly increased (P<0.05

P<0.01)

immobility duration and immobility frequency of model group were significantly decreased (P<0.05

P<0.01)

hippocampus cells was scattered

the expressions of CACNA1C and CaMK Ⅱ were decreased (P<0.05

P<0.01) in fluoxetine group and Shuyu capsule group. Conclusion: The rat model of PMDD liver-Qi stagnation can be successfully induced by forced swimming. Shuyu capsule may play a therapeutic role through Cav1.2-mediated CaM/CaMK Ⅱ signaling pathway and significantly improve the behavioral changes of rats with PMDD stagnation of liver-qi stagnation.

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