基于内质网应激探讨毛蕊异黄酮对慢性肾衰竭大鼠的影响

Effects of Calyx Isoflavones on Chronic Renal Failure in Rats Based on Endoplasmic Reticulum Stress

目的:探讨毛蕊异黄酮治疗慢性肾衰竭(CRF)大鼠的作用及其机制。方法:SPF级健康SD大鼠共48只,随机抽取8只为假手术组,其余40只为造模组,造模大鼠采用5/6肾切除建立大鼠CRF模型,造模成功后将造模大鼠随机分为模型组、开同组、毛蕊异黄酮低、中、高剂量组,开同组按1 g/kg灌胃,毛蕊异黄酮低、中、高剂量组分别按60、120、240 mg/kg灌胃,假手术组及模型组灌胃相应体积的生理盐水,每天1次,连续给药28天,采样。采用全自动血液分析仪检测血肌酐(Scr)、尿素氮(BUN)水平;H&E染色和Masson染色观察肾脏组织病理变化及纤维化程度;TUNEL法观察肾脏组织细胞凋亡;WB检测肾脏组织ATF6、Bax、caspase-3、CHOP、GRP78、Bcl-2蛋白表达。结果:毛蕊异黄酮可明显改善肾脏组织病理变化、纤维化程度及肾脏组织细胞凋亡;毛蕊异黄酮高剂量组较模型组可明显降低大鼠SCr、BUN水平及ATF6、Bax、caspase-3、CHOP、GRP78蛋白表达,明显升高Bcl-2蛋白表达(P<0.05)。结论:毛蕊异黄酮调控内质网应激,延缓肾损害进展,且高剂量组更有效。

Objective:To explore the effect and mechanism of calyx isoflavones in the treatment of chronic renal failure in(CRF)rats.Methods:A total of 48 healthy SD rats of SPF grade were randomly selected eight rats as sham operation group and the other 40 rats as model group,the model rats were established by 5/6 nephrectomy,after successful modeling,the rats were randomly divided into model group,Kaitong group,low-dose,middle-dose and high-dose groups of calyx isoflavones.The rats in the Kaitong group were given intragastric administration according to 1 g/kg,the low,middle and high dose groups of calyx isoflavones were intragastrically perfused with 60,120,and 240 mg/kg respectively,and the sham operation group and model group were intragastrically infused with the corresponding volume of normal saline,once a day for 28 days,and samples were collected.The levels of serum creatinine(Scr)and urea nitrogen(BUN)were detected by automatic hematology analyzer,the pathological changes of kidney tissue and the degree of renal fibrosis were observed by HE staining and Masson staining,the apoptosis of renal tissue was detected by TUNEL method,and the protein expressions of ATF6,Bax,caspase-3,CHOP,GRP78 and Bcl-2 in kidney tissue were detected by WB.Results:Calyx isoflavones could significantly improve the pathological changes,fibrosis degree and apoptosis of renal tissue,compared with the model group,the levels of Scr,BUN and the expression of ATF6,Bax,caspase-3,CHOP and GRP78 protein were significantly decreased and the expression of Bcl-2 protein was significantly increased in the high dose calyx isoflavone group.Conclusion:Calyx isoflavones regulate endoplasmic reticulum stress and delay the progression of renal damage,and the high dose group is more effective.