Celastrol attenuates sodium oxalate-induced acute kidney injury and crystal deposition by inhibiting NF-κB

LIU Yiheng; ZHENG Quanyou; ZHANG Wanyuan; YANG Chenhao; CHEN Siyu; LIN Wenbiao; ZHAO Siyu; XU Guilian; ZHANG Keqin

doi:10.16016/j.2097-0927.202501070

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Celastrol attenuates sodium oxalate-induced acute kidney injury and crystal deposition by inhibiting NF-κB

Basic Medicine | 更新时间：2025-04-14

- Celastrol attenuates sodium oxalate-induced acute kidney injury and crystal deposition by inhibiting NF-κB
- Journal of Army Medical University Vol. 47, Issue 7, Pages: 691-700(2025)
- 作者机构：
  
  1.重庆医科大学附属第二医院泌尿肾病中心，重庆
  2.；陆军军医大学（第三军医大学）：基础医学院免疫学教研室，重庆
  3.陆军军医大学（第三军医大学）：第一附属医院第九五八医院肾病泌尿科，重庆
- 作者简介：
  
  XU Guilian， E-mail： xuguilian2004@163.com
  ZHANG Keqin， E-mail： zhkq2004@163.com
- 基金信息：
  
  Founder： Supported by the General Project of Natural Science Foundation of Chongqing(CSTB2022NSCQ-MSX0099);the Scientific Research Cultivation Project for Undergraduates of Army Medical University(2024XBK24)
- DOI：10.16016/j.2097-0927.202501070
  CLC：
- Received：18 February 2025，
  
  Revised：02 March 2025，
  
  Published：15 April 2025
- 稿件说明：
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刘益亨,郑权友,张湾元,等.雷公藤红素通过抑制NF-κB减轻草酸钠诱导的急性肾损伤和晶体沉积[J].陆军军医大学学报,2025,47(07):691-700.

LIU Yiheng,ZHENG Quanyou,ZHANG Wanyuan,et al.Celastrol attenuates sodium oxalate-induced acute kidney injury and crystal deposition by inhibiting NF-κB[J].J Army Med Univ,2025,47(07):691-700.
刘益亨,郑权友,张湾元,等.雷公藤红素通过抑制NF-κB减轻草酸钠诱导的急性肾损伤和晶体沉积[J].陆军军医大学学报,2025,47(07):691-700. DOI：10.16016/j.2097-0927.202501070.

LIU Yiheng,ZHENG Quanyou,ZHANG Wanyuan,et al.Celastrol attenuates sodium oxalate-induced acute kidney injury and crystal deposition by inhibiting NF-κB[J].J Army Med Univ,2025,47(07):691-700. DOI：10.16016/j.2097-0927.202501070.

摘要

目的

探究雷公藤红素（Celastrol，Cel）在减轻草酸钠（sodium oxalate，NaOx）诱导的急性肾损伤（acute kidney injury，AKI）和晶体沉积中的作用及其机制。

方法

选用C57BL/6雄性小鼠（年龄8~12周，体质量22~24 g），按随机数字表法分为3组：Saline组（对照组）腹腔注射生理盐水，正常饮用水；NaOx组（损伤组），腹腔注射NaOx（75 mg/kg），NaOx溶液（50 μmol/L）作为饮用水；NaOx+Cel组（治疗组），提前24 h腹腔注射Cel（1 mg/kg），腹腔注射NaOx（75 mg/kg）， NaOx溶液（50 μmol/L）作为饮用水，24 h后收集所有标本。选用HK-2细胞构建模型，按随机数字表法分为4组：Medium组不作处理；NaOx组加入NaOx（终浓度500 μmol/L）处理细胞；NaOx+Cel组在预处理Cel（终浓度400 nmol/L）2 h后加入NaOx（终浓度500 μmol/L）处理细胞；NaOx+Cel+BA组在加入NaOx（终浓度500 μmol/L）2 h后加入NF-κB激动剂BA（终浓度8 μmol/L）处理细胞，其他步骤同NaOx+Cel组，24 h后收集各组细胞。采用Von Koosa和细胞黏附实验观察晶体沉积；采用HE染色观察肾脏组织病理并进行损伤评分；采用CCK-8检测NaOx和Cel的最适浓度；采集血清检测尿素和肌酐；采用IHC检测OPN、CD44、KIM-1、NGAL、p65、IL-1β、BAX、Caspase-3的表达；Western blot检测OPN、CD44、KIM-1、p65、P-p65、IL-1β蛋白表达。

结果

动物实验中，与NaOx组相比，NaOx+Cel组小鼠晶体沉积减少（

0.000 1），肾小管受损减轻（

0.01），血清尿素和肌酐降低（

0.05），肾脏黏附分子OPN与CD44、肾脏损伤分子KIM-1与NGAL、炎症相关分子p65与IL-1β、凋亡相关分子BAX、Caspase-3表达均下降（

0.05）。而在细胞实验中，与NaOx组相比，NaOx+Cel组的晶体黏附减少（

0.000 1），黏附分子OPN与CD44表达下降（

0.05），炎症分子IL-1β和P-p65/p65的比值下调（

0.05），肾损伤分子KIM-1下调（

0.05）。与NaOx+Cel组相比，NaOx+Cel+BA组晶体黏附明显增多（

0.000 1），炎症分子IL-1β和P-p65/p65的比值上升（

0.05），肾损伤分子KIM-1增多（

0.05）。

结论

雷公藤红素可能通过抑制NF-κB的活化减轻NaOx诱导的肾组织中晶体的沉积和AKI。

Abstract

Objective‍　‍To investigate the role and possible mechanism of celastrol (Cel) in sodium oxalate (NaOx)-induced acute kidney injury (AKI) and crystal deposition in the kidney tissues in mice. Methods‍　‍Male C57BL/6 mice (aged 8~12 weeks

weighing 22~24 g) were randomly divided into 3 groups. Saline group (control group

intraperitoneal injection with normal saline and drinking water freely)

NaOx group (injured group

intraperitoneal injection of 75 mg/kg NaOx

and drinking water containing 50 μmol/L NaOx)

and NaOx+Cel group (treatment group

intraperitoneal injection of 1 mg/kg Cel firstly and then 75 mg/kg NaOx in 24 h later

drinking water containing 50 μmol/L NaOx). All specimens were collected in 24 h after NaOx injection. HK-2 cells were randomly divided into 4 groups: Medium group (no treatment)

NaOx group (500 μmol/L NaOx)

NaOx+Cel group (400 nmol/L Cel pre-treatment for 2 h followed by 500 μmol/L NaOx treatment)

and NaOx+Cel+BA group [8 μmol/L betulinic acid (BA

NF-κB agonist) after the interventions as the NaOx+Cel group

]

. Cells of each group were collected in 24 h after corresponding treatments. Von Koosa and cell adhesion assays were used to observe crystal deposition. HE staining was employed to observe renal histopathology and score the damage. CCK-8 assay was utilized to detect cell viability to obtain the optimal concentrations of NaOx and Cel. Serum urea and creatinine levels were detected. Immunohisotochemical assay was conducted to detect the expression of OPN

CD44

KIM-1

NGAL

p65

IL-1β

BAX

and Caspase-3

and Western blotting was performed for protein levels of OPN

CD44

KIM-1

p65

P-p65 and IL-1β. Results‍　‍The mice in the NaOx+Cel group showed reduced crystal deposition (

0.0001)

attenuated renal tubular damage (

0.01)

decreased serum urea and creatinine levels (

0.05)

and declined expression levels of the renal adhesion molecules OPN and CD44

the kidney injury molecules KIM-1 and NGAL

the inflammation-associated molecules p65 and IL-1β

and the apoptosis related molecules BAX and Caspase-3 when compared with the NaOx group (

0.05). In

in vitro

study

the NaOx+Cel group showed reduced crystal adhesion (

0.0001)

decreased expression of the adhesion molecules OPN and CD44 (

0.05)

down-regulation of the inflammatory molecule IL-1β and P-p65/p65 ratio (

0.05)

and down-regulation of the renal injury molecule KIM-1 (

0.05) when compared with the NaOx group. In the NaOx+Cel+BA group

crystal adhesion was significantly increased (

0.0001)

the inflammatory molecule IL-1β and the ratio of P-p65/p65 were increased (

0.05)

and the kidney injury molecule KIM-1 was increased when compared with the NaOx+Cel group (

0.05). Conclusion‍　‍Cel may reduce NaOx-induced crystal deposition and AKI by inhibiting NF-κB activation.

关键词

Keywords

references

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