Effects of Shenshuaikang Enema Liquid on the Wnt/β-catenin pathway for inhibiting the hypoxia reoxygenation injury in HK-2 cells
10.3969/j.issn.1001-1528.2018.05.006
- VernacularTitle:Wnt/β-catenin通路在肾衰康灌肠液抑制HK-2细胞缺氧/复氧损伤的作用
- Author:
Rong-Rong DENG
1
;
Xin-Yi LUO
;
Nai-Jing YE
;
Xin-Ping WU
;
Yi-Jun WANG
;
Ming-Quan LI
Author Information
1. 成都中医药大学,四川成都610075
- Keywords:
Shenshuaikang Enema Liquid;
hypoxia/reoxygenation;
H/R;
HK-2;
Wnt/β-catenin
- From:
Chinese Traditional Patent Medicine
2018;40(5):1036-1042
- CountryChina
- Language:Chinese
-
Abstract:
AIM To investigate the protective effect of serum containing Shenshuaikang Enema Liquid (Rhei Radix et Rhizoma,Salviae miltiorrhizae Radix et Rhizoma,Carthami Flos,Astragali Radix) on HK-2 cells injured by hypoxia/reoxygenation (H/R) and its effect on Wnt/β-catenin pathway.MEHTODS Six New Zealand white rabbits randomly divided into control group,Shenshuaikang high dose group and PBS group (n =2) were treated accordingly for serum collection after 3 days' enema.The HK-2 cells injured by hypoxia/reoxygenation were then administered with serum of rabbits from the control group,PBS group,Shenshuaikang groups (high dose,middle dose and low dose groups due to the differently diluted concentrations) respectively.The H/R damage was determined by ROS fluorescence probe,the cellular damage/apoptosis were analyzed by CFSE/PI method and flow cytometry combined with Annexin V-FITC/PI,and the investigation on effects of drugs on expression of Wnt4 mRNA and β-catenin mRNA were accomplished by fluorescence quantitative PCR.RESULTS The fluorescence intensity of intracellular ROS expression was significantly increased after modeling.CFSE/PI double staining showed that the variant Shenshuaikang dose groups displayed obvious proportional mortality superiority to either the control group or PBS group,and the high dose group achieved the lowest mortality.Annexin V-FITC/PI and flow cytometry showed that,at 12 h,compared with the control group (45.6 ± 2.2)％ and PBS group (41.6 ±0.7) ％,Shenshuaikang groups [low dose group (14.8 ± 0.3) ％,middle dose group (10.3 ± 0.6) ％,high dose group (12.9 ± 0.9)％] obviously inhibited apoptosis/death.Shenshuaikang Enema Liquid medicated serum demonstrated its significant effect on the increase of the Wnt4 mRNA expression and a dual-directional regulation on the expression of β-caterin mRNA by quantitative PCR (P ＜ 0.05).CONCLUSION Inhibition of the apoptosis/death of HK-2 cells with hypoxia/reoxygenation injury due to the serum containing Shenshuaikang Enema Liquid suggests the agent's influence on the activation of Wnt/β-catenin pathway.
