中国中西医结合杂志2025，Vol.45Issue(8)：952-960,9.DOI:10.7661/j.cjim.20250503.079
狼疮定合并激素抑制eIF2α-ATF4-CHOP内质网应激信号通路治疗狼疮性肾炎小鼠
Langchuangding Combined with Glucocorticoids Treats Lupus Nephritis via Inhibiting in MRL/Ipr Mice Based on the eIF2α-ATF4-CHOP Endoplasmic Reticulum Stress Signaling Pathway
摘要
Abstract
Objective To reveal the mechanisms for Langchuangding(LCD)combined with glucocorticoids(GCs)in the treatment of lupus nephritis(LN),based on the endoplasmic reticulum(ER)stress pathway involving eukaryotic translation-initiation factor 2α(eIF2α),activating transcription factor 4(ATF4),and C/EBP homologous protein(CHOP)in renal tissue of MRL/Ipr mice.Methods Six-week-old female MRL/Ipr model mice were randomly divided into four groups,including model,GCs,LCD,and LCD combined with GCs groups(n=10).Female MRL/MpJ mice were used as the control(n=10).The mice were sacrificed at 8 and 14 weeks,respectively.The pathological morphological changes were detected by HE staining,and the deposition of complement C3 and IgG in renal tissues were also determined by immunofluorescence and immunohistochemical method.The expression levels of ER stress marker proteins,such as glucose regulated protein(GRP78),eIF2α,p-eIF2α,ATF4 and CHOP,and their mRNA were measured by Western Blot and RT-qPCR analysis,respectively.Results In addition to no significant pathological changes,no obvious deposition of complement C3 and IgG was observed in the kidney of 8-week-old model mice compared with the normal group,while at 14 weeks,there existed significant complement C3 and IgG deposition,noticeable damages in glomeruli,interstitium,and blood vessels,and extensive pathological changes(e.g.,inflammatory cell infiltration).However,the expression levels of eIF2α,p-eIF2α,ATF4,and CHOP proteins,and the mRNA levels of GRP78,eIF2α,ATF4,and CHOP significantly increased in the renal tissue of both 8-and 14-week-old model mice(P＜0.01).Compared with the 8-week model group,the levels of eIF2α and ATF4 protein,and ATF4 mRNA were reduced in GCs group(P＜0.05,P＜0.01).The levels of eIF2α,ATF4,and CHOP proteins reduced in LCD group(P＜0.05,P＜0.01).Meanwhile the levels of eIF2α,p-eIF2α,ATF4,CHOP(P＜0.01)and the level of GRP78 and ATF4 mRNA(P＜0.05)decreased in LCD combined with GCs group.At 14 weeks,accompanying with the reduced deposition of immune complexes in renal tissues,the pathological changes,including glomerular swelling and inflammatory cell infiltration,were effectively alleviated in all of the intervention groups.Compared with the 14-week model group,the levels of GRP78,eIF2α,p-eIF2α,ATF4,CHOP proteins and their mRNA decreased in LCD combined with GCs group(P＜0.05,P＜0.01).The proteins levels of GRP78,p-eIF2α,ATF4,CHOP and the levels of GRP78,ATF4,CHOP mRNA reduced in both GCs and LCD groups(P＜0.01).Furthermore,compared with GCs group,the expression of ATF4 protein and GRP78 mRNA reduced in LCD combined with GCs group at 8 weeks of age(P＜0.01);while at 14 weeks,LCD combined with GCs group displayed the lower pathological damage score and the reduced deposition of complement C3 and IgG(P＜0.05,P＜0.01).Compared with LCD group,the expressions of p-eIF2α protein and ATF4 mRNA reduced in both GCs and LCD combined with GCs groups at 8 weeks,and the expression level of GRP78 mRNA in the combined group also decreased(P＜0.01);at 14 weeks,GCs group showed decreased expressions of ATF4 and CHOP proteins as well as eIF2α mRNA level,while LCD combined with GCs group displayed reduced deposition of complement C3 and IgG,and the much lower expressions of CHOP protein,and GRP78,eIF2α,ATF4 mRNA(P＜0.05,P＜0.01).Conclusions ER stress was already present in the early stage of LN and worsened with disease progression.LCD combined with GCs treatment could significantly relieve ER stress,effectively alleviating LN.Moreover,its therapeutic effect was better than glucocorticoid treatment.关键词
狼疮肾炎/内质网应激/狼疮定/糖皮质激素/中西医结合Key words
lupus nephritis/endoplasmic reticulum stress/Langchuangding/glucocorticoid/integrative medicine引用本文复制引用
陈艺霖,许小芬,卢璐,谢冠群,杜羽,胡长锋..狼疮定合并激素抑制eIF2α-ATF4-CHOP内质网应激信号通路治疗狼疮性肾炎小鼠[J].中国中西医结合杂志,2025,45(8):952-960,9.基金项目
国家自然科学基金资助项目(No.82274230,No.82104849) （No.82274230,No.82104849）
浙江省自然科学基金资助项目(No.LQ23H290002) （No.LQ23H290002）
