The hemostatic effect and its mechanism of modified Sijunzi Decoction on the zebrafish intestinal bleeding model through 5-hydroxytryptamine and its receptors
10.3969/j.issn.1006-2157.2024.04.012
- VernacularTitle:加味四君子汤通过影响5-羟色胺及其受体对斑马鱼肠道出血模型的止血效应及其机制探讨
- Author:
Jun WANG
1
;
Haiyan LANG
;
Jiali ZHOU
;
Xueying ZHANG
;
Yanhong KANG
;
Yun ZHANG
;
Xinyi CHEN
;
Wei MA
Author Information
1. 浙江中医药大学附属第一医院(浙江省中医院) 杭州 310000
- Keywords:
immune thrombocytopenia;
modified Sijunzi Decoction;
5-hydroxytryptamine;
5-hydroxytryptamine receptors;
protein kinase B;
extracellular signal-regulated protein kinase;
zebrafish
- From:
Journal of Beijing University of Traditional Chinese Medicine
2024;47(4):516-522
- CountryChina
- Language:Chinese
-
Abstract:
Objective We aimed to investigate the hemostatic effect and mechanism of modified Sijunzi Decoction(MSJZT),consisting of tangshen,India bread,largehead atractylodes rhizome,liquorice root,milkvetch root,ass hide glue,and India madder root,on the zebrafish intestinal bleeding model through 5-hydroxytryptamine(5-HT)and its receptors(5-HT2aR,5-HT2bR).Methods Zebrafish at 4 days post fertilization were used as the research object.An intestinal bleeding model was established by induction with 6 μmol/L simvastatin for 24 hours.The zebrafish were divided into normal group,model group,MSJZT low-,mid-and high-concentration groups(55.6,167,500 mg/L),and after modeling,the corresponding concentration of MSJZT was administered.The samples were collected after 24 hours.Platelet count(PLT)and bleeding status were observed.The content of 5-HT and Caspase 3/7 activity were detected by colorimetry.The gene expressions of 5-HT2aR,5-HT2bR and serotonin transporter(SERT)were detected by real-time PCR.The protein expressions of protein kinase B(AKT),phosphorylated AKT(p-AKT),extracellular signal-regulated protein kinase(ERK),and phosphorylated ERK(p-ERK)were detected by Western blotting.Results The intestinal bleeding rate in the model group was 70.0%,and that in the MSJZT low-,mid-and high-concentration groups was 36.7%,40.0%,and 80.0%,respectively;the intestinal hemostatic effect was 54%,52%,and 7%,respectively.Compared with the model group,the PLT in the MSJZT low-and mid-concentration groups decreased;the content of 5-HT in all MSJZT groups increased,and the gene expressions of 5-HT2aR and 5-HT2bR were up-regulated.The gene expression of SERT was up-regulated in the low-and mid-concentration groups;the Caspase 3/7 activity in the mid-and high-concentration groups was reduced;the expressions of p-AKT and p-ERK proteins were up-regulated in the low-and mid-concentration groups(all P<0.05).Conclusion MSJZT can reduce the incidence and severity of intestinal bleeding in zebrafish intestinal bleeding models.The mechanism may be achieved by activating AKT and ERK signaling pathways through 5-HT and its receptors.
