Effect of Serum Containing <italic style="font-style: italic">Zhizhuwan Decoction</italic> （枳术丸汤剂）on Proliferation and Colonic Interstitial Cells Cajal and PLC-γ Signaling Pathway in Rats

Xuting XIA; Ting WANG; Fulin LIU; Qinrong TU; Guangfei TENG

doi:10.13288/j.11-2166/r.2021.21.014

您当前的位置：

首页 >

文章列表页 >

Effect of Serum Containing Zhizhuwan Decoction （枳术丸汤剂）on Proliferation and Colonic Interstitial Cells Cajal and PLC-γ Signaling Pathway in Rats

更新时间：2021-11-02

- Effect of Serum Containing Zhizhuwan Decoction （枳术丸汤剂）on Proliferation and Colonic Interstitial Cells Cajal and PLC-γ Signaling Pathway in Rats
- Journal of Traditional Chinese Medicine Vol. 62, Issue 21, Pages: 1923-1929(2021)
- 作者机构：
  
  1.湖南中医药大学，湖南省长沙市岳麓区含浦科教园学士路300号，410208
  2.湖北省襄阳市中医医院
  3.湖南师范大学附属湘东医院
  4.中国中医科学院望京医院
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2021.21.014
  CLC：
- Received：23 May 2021，
  
  Revised：2021-08-07，
  
  Published：02 November 2021
- 稿件说明：
移动端阅览
夏旭婷,王婷,刘富林等.枳术丸汤剂含药血清对大鼠结肠Cajal间质细胞增殖及PLC-γ信号通路的影响[J].中医杂志,2021,62(21):1923-1929.

XIA Xuting,WANG Ting,LIU Fulin,et al.Effect of Serum Containing Zhizhuwan Decoction （枳术丸汤剂）on Proliferation and Colonic Interstitial Cells Cajal and PLC-γ Signaling Pathway in Rats[J].Journal of Traditional Chinese Medicine,2021,62(21):1923-1929.
夏旭婷,王婷,刘富林等.枳术丸汤剂含药血清对大鼠结肠Cajal间质细胞增殖及PLC-γ信号通路的影响[J].中医杂志,2021,62(21):1923-1929. DOI： 10.13288/j.11-2166/r.2021.21.014.

XIA Xuting,WANG Ting,LIU Fulin,et al.Effect of Serum Containing Zhizhuwan Decoction （枳术丸汤剂）on Proliferation and Colonic Interstitial Cells Cajal and PLC-γ Signaling Pathway in Rats[J].Journal of Traditional Chinese Medicine,2021,62(21):1923-1929. DOI： 10.13288/j.11-2166/r.2021.21.014.

摘要

目的

探讨枳术丸治疗慢传输型便秘的可能作用机制。

方法

20只SD雄性大鼠随机分为枳术丸汤剂组、空白组，每组10只。枳术丸汤剂组大鼠每日给予浓度为2 g/ml枳术丸汤剂5.8 g灌胃，连续7天，空白组正常饲养。末次给药后1 h腹主动脉采血离心制备含药血清。采用CCK8法观察磷脂酶C-γ（PLC-γ）特异性抑制剂U73122对大鼠结肠原代Cajal间质细胞的细胞抑制率来筛选药物浓度，观察5%、10%、15%、20%浓度的枳术丸汤剂含药血清对Cajal间质细胞的细胞增殖率筛选药物浓度，并分别培养12、24、48 h来筛选用药时间。取二或三代Cajal间质细胞接种至96孔板中，每孔1×10

个细胞，每组重复3孔。实验分为正常组、模型组（正常细胞+筛选浓度的U73122）、空白血清组（模型组+10%空白血清）、枳术丸汤剂血清组（模型组+筛选浓度枳术丸含药血清）。干预时间为实验筛选出的时间。免疫组化法检测磷脂酶C-γ1（PLC-γ1）、磷脂酶C-γ2（PLC-γ2）阳性表达，Western blot法检测PLC-γ1、PLC-γ2蛋白表达，RT-PCR法检测PLC-γ1、PLC-γ2基因表达。

结果

筛选出U73122最佳实验药物浓度为30 μmol/L，枳术丸汤剂组最佳用药浓度为5%，实验最佳干预时间为24 h。与正常组比较，模型组PLC-γ1、PLC-γ2平均光密度值、蛋白及基因表达均降低（

＜0.01）；与模型组比较，枳术丸汤剂血清组、空白血清组PLC-γ1、PLC-γ2平均光密度值、蛋白及基因表达均升高（

＜0.05或

＜0.01）；与空白血清组比较，枳术丸汤剂血清组PLC-γ1、PLC-γ2阳性表达、蛋白及基因表达明显增多（

＜0.01）。

结论

枳术丸汤剂可能通过激活Cajal间质细胞PLC-γ信号通路，调控细胞增殖，从而促进胃肠运动。

Abstract

Objective

To explore the possible mechanism of

Zhizhuwan Decoction

（枳术丸汤剂， ZD） for treatment of slow transit constipation （STC）.

Methods

Twenty SD male rats were randomized into ZD group and blank group， with 10 rats in each group. The rats in the ZD group were given 5.8 g ZD by gavage at a concentration of 2 g/ml per day for seven consecutive days， while those in the blank group were fed normally. One hour after the last administration， blood was collected from the abdominal aorta and centrifuged to prepare the medicated serum. CCK8 method was used to assess the cell inhibition rate of U73122， a specific inhibitor of phospholipase C-γ （PLC-γ），on primary interstitial cells of Cajal （ICC）； the concentration of medicated serum was selected from 5%， 10%， 15% and 20% by observing the effect on cell proliferation of ICC； the medication time was selected from 12 h， 24 h and 48 h based on the results. Second or third generation of ICC were seeded into a 96-well plate， with 1×10

cells per well and three repeated wells for each of the three groups， which were normal group， model group （normal cells plus U73122 at the selected concentration）， blank serum group （model group plus 10% blank serum） and ZD-containing serum group （model group plus ZD-containing serum at the selected concentration）； the administrations lasted for the selected time duration. The positive expression of PLC-γ 1 and PLC-γ 2 were detected by immunohistochemistry， and the protein and gene expression of PLC-γ 1 and PLC-γ 2 were detected through Western blotting and RT-PCR， respectively.

Results

The selected best concentration was 30 μmol/L of U73122， and was 5% of ZD； and the selected administration time was 24 h. Compared to the normal group， the model group had decreased PLC-γ1， PLC-γ2 average optical density， protein and gene expression （

＜0.01）； compared to the model group， the ZD-containing serum group and blank serum group had higher PLC-γ1 and PLC-γ2 average optical density， protein and gene expression （

＜0.05 or

＜0.01）； compared to the blank serum group， the ZD-containing serum group had significantly increased positive expression of PLC-γ1， PLC-γ2， as well as protein and gene expression （

＜0.01）.

Conclusion

ZD may regulate cell proliferation through activation of the PLC-γ signaling pathway of ICC， so as to promote the gastrointestinal motility.

关键词

Keywords

references

GARCIA-LOPEZ P ， GARCIA-MARIN V ， MARTINEZ-MURILLO R ， et al . Updating old ideas and recent advances regarding the interstitial cells of Cajal ［J］. Brain Res Rev ， 2009 ， 61 （ 2 ）： 154 - 169 .

REBECCHI MJ ， PENTYALA SN . Structure，function，and control of phosphoinositide-specific phospholipase C ［J］. Physiol Rev ， 2000 ， 80 （ 4 ）： 1291 - 1335 .

刘富林，易健，刘柏炎，等 . 超微枳术丸对功能性消化不良小鼠胃肠运动及MTL、GAS的影响［J］. 中国中医急症， 2011 ， 20 （ 4 ）： 594 - 595，604 .

滕广飞，刘富林，夏旭婷，等 . 枳术丸水煎液及其拆方含药血清对大鼠结肠Cajal间质细胞增殖和凋亡的影响［J］. 中国实验方剂学杂志， 2020 ， 26 （ 19 ）： 120 - 126 .

王伟松，夏旭婷，刘富林，等 . 枳术丸对脾虚证慢传输型便秘小鼠肠道运动及PLC-γ1/PLC-γ2信号通路的影响［J］. 中国实验方剂学杂志， 2019 ， 25 （ 10 ）： 8 - 14 .

王浩 . 枳术丸加减治疗老年慢传输型便秘的临床研究［D］. 南京：南京中医药大学， 2019 .

夏旭婷，刘富林，王文凤，等 . 枳术丸对慢传输型便秘脾虚证模型小鼠肠道传导功能的影响［J］. 中医杂志， 2020 ， 61 （ 19 ）： 1725 - 1730 .

夏旭婷，樊俊阳，易健，等 . Cajal间质细胞在慢传输型便秘发病机制中的作用［J］. 湖南中医杂志， 2014 ， 30 （ 10 ）： 172 - 174 .

张旭，徐华，毕淑英，等 . 大鼠胃肠道不同部位Cajal间质细胞与慢传输型便秘的关系［J］. 胃肠病学和肝病学杂志， 2012 ， 21 （ 6 ）： 536 - 540 .

JI QS ， WINNIER GE ， NISWENDER KD ， et al . Essential role of the tyrosine kinase substrate phospholipase C-gamma1 in mammalian growth and development ［J］. Proc Natl Acad Sci U S A ， 1997 ， 94 （ 7 ）： 2999 - 3003 .

孟凡华，葛彦明，王素霞，等 . 系统性红斑狼疮的磷脂酶C-γ的致病机制及靶向治疗研究进展［J］. 实用医药杂志， 2017 ， 34 （ 3 ）： 269 - 271 .

DIENI CA ， STOREY KB . Protein kinase C in the wood frog，Rana sylvatica：reassessing the tissue-specific regulation of PKC isozymes during freezing ［J］. Peer J ， 2014 ， 2 ： e558 . doi： 10.7717/peerj.558. eCollection 2014 http://dx.doi.org/10.7717/peerj.558.eCollection2014 .

查看原文

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Effect of Tongbian Decoction （通便汤） on the VAPB-PTPIP51 Complex and Autophagy of Interstitial Cells of Cajal in the Colon of Slow Transit Constipation Model Rats

Study on the Effect of Intestinal Flora on Intestinal Motility in Rats with Slow Transit Constipation of Qi Stagnation Pattern Based on Pseudo-Sterile Experiment and Fecal Microbiota Transplantation

Clinical Observation on Huangxin Runchang Pian（黄杏润肠片） in Treating 45 Cases of Mild or Moderate Slow Transit Constipation

Comparison of Effect of Serum Containing Yiqi Yangxue Decoction（益气养血汤） with Hongqi（Radix Hedysari红芪） and the Same Serum with Huangqi（Radix Astragali,黄芪） on T-Lymphocytes in Spleen of SAMP8 Mice

Clinical Observation on Baliao Acupoints Catgut Embedding for 30 Cases of Slow Transit Constipation

Related Author

WANG Chuyue

LI Jiacheng

YANG Yingqi

SHEN Sicheng

CHEN Zhiyang

XU Zhizhong

WU Bensheng

CHEN Meiyao

Related Institution

Suzhou Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Chinese Medicine

Fujian Provincial Key Laboratory of Chinese Medicine Preparation in Medical Institutions

Second People's Hospital Affiliated to Fujian University of Traditional Chinese Medicine

Shuguang Hospital,Affiliated to Shanghai University of Chinese Medicine

School of pharmacy,Lanzhou University

Address：No.16, Nanxiao street, Dongzhimen, Dongcheng District, Beijing Postal code：100700
Tel：010-64035632 Email：jtcmcn@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备09064830号-19 京公网安备11010102007558号
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰

Effect of Serum Containing Zhizhuwan Decoction （枳术丸汤剂）on Proliferation and Colonic Interstitial Cells Cajal and PLC-γ Signaling Pathway in Rats

DOI：10.13288/j.11-2166/r.2021.21.014

摘要

Abstract

关键词

Keywords

references