温阳活血利水方含药血清对足细胞组织蛋白酶L、发动蛋白以及微丝、微管的影响

陈凤; 王相相; 邱雯雯; 袁军

doi:10.13288/j.11-2166/r.2018.10.016

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温阳活血利水方含药血清对足细胞组织蛋白酶L、发动蛋白以及微丝、微管的影响

实验研究 | 更新时间：2021-11-10

- 温阳活血利水方含药血清对足细胞组织蛋白酶L、发动蛋白以及微丝、微管的影响
- Effect of Serum Containing Wenyang Huoxue Lishui Formula（温阳活血利水方） on Podocytes Cathepsin L,Dynamin,Microfilament and Microtubule
- 中医杂志 2018年59卷第10期页码：882-886
- 作者机构：
  
  湖北中医药大学
- 作者简介：
- 基金信息：
  
  湖北省卫生计生委中医药中西医结合科研项目
- DOI：10.13288/j.11-2166/r.2018.10.016
  中图分类号： R285.5
- 纸质出版：2018
- 稿件说明：
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[1]陈凤,王相相,邱雯雯,袁军.温阳活血利水方含药血清对足细胞组织蛋白酶L、发动蛋白以及微丝、微管的影响[J].中医杂志,2018,59(10):882-886.

CHEN Feng, WANG Xiangxiang, QIU Wenwen, et al. Effect of Serum Containing Wenyang Huoxue Lishui Formula（温阳活血利水方） on Podocytes Cathepsin L,Dynamin,Microfilament and Microtubule[J]. Journal of Traditional Chinese Medicine2018, 59(10): 882-886.
[1]陈凤,王相相,邱雯雯,袁军.温阳活血利水方含药血清对足细胞组织蛋白酶L、发动蛋白以及微丝、微管的影响[J].中医杂志,2018,59(10):882-886. DOI： 10.13288/j.11-2166/r.2018.10.016.

CHEN Feng, WANG Xiangxiang, QIU Wenwen, et al. Effect of Serum Containing Wenyang Huoxue Lishui Formula（温阳活血利水方） on Podocytes Cathepsin L,Dynamin,Microfilament and Microtubule[J]. Journal of Traditional Chinese Medicine2018, 59(10): 882-886. DOI： 10.13288/j.11-2166/r.2018.10.016.

摘要

目的探讨温阳活血利水方治疗原发性肾病综合征蛋白尿的可能作用机制。方法将足细胞分为正常对照组、模型组、地塞米松组、10%中药血清组、20%中药血清组、空白对照组。除正常对照组、空白对照组外

其余各组用氨基核苷嘌呤霉素粉剂制备足细胞损伤模型。正常对照组和模型组加入完全培养基5 ml

地塞米松组加入392μg/L地塞米松5 ml

10%中药血清组、20%中药血清组、空白对照组分别加入10%中药血清、20%中药血清、空白对照血清各5 ml。RT-PCR法与Western blot分别检测各组足细胞胞浆组织蛋白酶L（CatL）、发动蛋白mRNA和蛋白表达含量

并用激光共聚焦观察各组中足细胞骨架微丝、微管的形态结构变化。结果与正常组比较

模型组CatL mRNA与蛋白含量升高

发动蛋白表达含量降低（P<0.01）;与模型组比较

各中药血清组与地塞米松组CatL mRNA与蛋白表达含量降低

足细胞发动蛋白蛋白表达含量升高（P<0.01）;与10%中药血清组比较

20%中药血清组CatL mRNA与蛋白表达含量降低

发动蛋白表达含量升高（P<0.01）。各组间足细胞发动蛋白mRNA表达含量差异无统计学意义（P>0.05）。结论温阳活血利水方可能通过下调足细胞CatL的表达

减少CatL对发动蛋白的酶解

从而稳定细胞骨架中的微丝、微管等结构

减少足突融合

改善蛋白尿。

Abstract

Objective To explore the possible mechanism of Wenyang Huoxue Lishui Formula-（ warming yang

activating blood circulation

promoting urination decoction

温阳活血利水方） in treating proteinuria of primary nephrotic syndrome. Methods Mouse podocytes were divided into normal control group

model group

dexamethasone group

10% Chinese medicine serum group

20% Chinese medicine serum group

and blank control group. Except for normal control group

blank control group

the other groups were given puromycin aminonucleoside powder to induce podocytes injury model. The normal control group and model group were added 5 ml

and dexamethasone group was added 5 ml dexamethasone 392 μg/L. 10% Chinese medicine serum group

20% Chinese medicine serum group

and blank control group were added 5 ml 10% Chinese medicine serum

20% Chinese medicine serum and blank control serum respectively. RT-PCR and Western blot were used to detect the protein expression of cathepsin L（ CatL）and dynamin mRNA

and laser confocal was applied to observe the morphological and structural changes of microfilaments and microtubules in the podocytes of each group. Results Compared with the normal group

the mRNA and protein expression of CatL of model group increased and the protein expression of dynaminin decreased（ all P <0. 01）. Compared with the model group

the mRNA and protein expression of CatL in Chinese medicine serum groups and dexamethasone group decreased

and the expression of dynamin was increased（ all P < 0. 01）. Compared with the treatment with the serum contained 10% Chinese medicine

the mRNA and protein expression of CatL were lower and the protein expression of dynamin was higher by treatment with the serum contained 20% Chinese medicine（ P <0. 01）. There was no significant difference in the relative expression of dynamin mRNA among the groups（ P >0. 05）. Conclusion Wenyang Huoxue Lishui Formula can reduce CatL’s enzymolysis of dynamin through reducing CatL expression in podocytes

so as to stabilize microfilaments

microtubules and other structures in the cytoskeleton

reduce fusion of podocytic process and improve proteinuria.

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