Protective Effect of Tongluo Baoshen Prescription-containing Serum on Lipopolysaccharide-induced Podocyte Injury of Rats
10.13422/j.cnki.syfjx.20250497
- VernacularTitle:通络保肾方含药血清对脂多糖诱导大鼠足细胞损伤的保护作用
- Author:
Yongfang LIU
1
;
Tiantian YIN
2
;
Huiyang LIU
1
;
Rui HUANG
2
;
Zhiying FENG
2
;
Li ZHOU
2
Author Information
1. Jiujiang Hospital of Traditional Chinese Medicine，Jiangxi University of Chinese Medicine，Jiujiang 332000，China
2. Graduate School，Jiangxi University of Chinese medicine，Nanchang 330004，China
- Publication Type:Journal Article
- Keywords:
Tongluo Baoshen prescription;
immunoglobulin A （IgA） nephropathy;
podocyte;
pyroptosis
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2025;31(10):139-148
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo observe the effects of Tongluo Baoshen prescription （TLBS）-containing serum on the rat podocyte injury induced by lipopolysaccharide （LPS） and explore the potential mechanisms. MethodsSD rats were used to prepare the blank serum， losartan potassium-containing serum， and low-， medium-， and high-dose TLBS-containing sera. Rat podocytes were cultured in vitro， and the effects of drug-containing sera on podocyte viability were detected by the cell counting kit-8 （CKK-8） method. The optimal intervention volume fraction of drug-containing sera and the optimal concentration of LPS for inducing the podocyte injury were determined. Rat podocytes were grouped as follows： normal control （NC， 10% blank serum）， model control （MC， 20.00 mg·L-1 LPS+10% black serum）， losartan potassium （LP， 20.00 mg·L-1 LPS+10% losartan potassium-containing serum）， low-dose TLBS （TLBS-L， 20.00 mg·L-1 LPS+10% low-dose TLBS-containing serum）， medium-dose TLBS （TLBS-M， 20.00 mg·L-1 LPS+10% medium-dose TLBS-containing serum）， and high-dose TLBS （TLBS-H， 20.00 mg·L-1 LPS+10% high-dose TLBS-containing serum）， and the interventions lasted for 48 h. The ultrastructure of podocytes was observed under a transmission electron microscope. The podocyte apoptosis was detected by the terminal deoxynucleoitidyl transferase mediated nick-end labeling （TUNEL） kit. Immunofluorescence was used to detect the expression of gasdermin D N-terminal fragment （GSDMD-NT） in podocytes. The mRNA and protein levels of G protein-coupled receptor family C group 5 member B （GPRC5B）， nuclear factor-κB （NF-κB） p50， NF-κB p52， NF-κB p65， Rel B， c-Rel， NOD-like receptor protein 3 （NLRP3）， cysteinyl aspartate-specific protease-1 （Caspase-1）， GSDMD-NT， interleukin （IL）-1β， IL-18， nephrin， integrin α3， and integrin β1 in podocytes were determined by real-time quaritiative polymerase chain reaction （Real-time PCR） and Western blot， respectively. ResultsCompared with the NC group， the MC group showed reduced podocyte protrusions and organelles， segmental missing of cell membranes， increased and swollen mitochondria， irregular nuclear membranes， light chromatin， increased TUNEL fluorescence-positive nuclei （P<0.01）， obviously enhanced fluorescence intensity of GSDMD-NT， up-regulated mRNA and protein levels of GPRC5B， NF-κB p50， NF-κB p52， NF-κB p65， Rel B， c-Rel， NLRP3， caspase-1， GSDMD-NT， IL-1β， and IL-18 （P<0.01）， and down-regulated mRNA and protein levels of nephrin， integrin α3， and integrin β1 （P<0.01） in podocytes. Compared with the MC group， the LP， TLBS-M， and TLBS-H groups showed improved ultrastructure of podocytes with increased protrusions， intact cell membranes， reduced organelles， and alleviated mitochondrial swelling， decreased TUNEL fluorescence-positive nuclei （P<0.01）， weakened fluorescence intensity of GSDMD-NT， down-regulated mRNA and protein levels of GPRC5B， NF-κB p50， NF-κB p52， NF-κB p65， Rel B， c-Rel， NLRP3， caspase-1， GSDMD-NT， IL-1β， and IL-18 （P<0.01）， and up-regulated mRNA and protein levels of nephrin， integrin α3， and integrin β1 （P<0.05， P<0.01）. Moreover， the changes above were the most obvious in the TLBS-H group. ConclusionThe TLBS-containing serum can regulate the GPRC5B/NF-κB/NLRP3 pathway to inhibit pyroptosis， thereby ameliorating the podocyte injury induced by LPS.
