目的探究内质网应激(ERS)相关凋亡通路在单侧输尿管梗阻(UUO)大鼠肾间质纤维化中的作用机制。方法将大鼠随机分为假手术组及UUO模型组,UUO组行左侧输尿管结扎术,于术后3、7和14 d HE和Masson染色观察肾脏病理变化;眼底静脉丛取血,分离...目的探究内质网应激(ERS)相关凋亡通路在单侧输尿管梗阻(UUO)大鼠肾间质纤维化中的作用机制。方法将大鼠随机分为假手术组及UUO模型组,UUO组行左侧输尿管结扎术,于术后3、7和14 d HE和Masson染色观察肾脏病理变化;眼底静脉丛取血,分离血清测定血尿素氮及肌酐;Western blot检测葡萄糖调节蛋白78(GRP78)、内质网源性转录因子(CHOP)、凋亡相关蛋白半胱氨酸天门冬氨酸蛋白酶3(caspase-3)及caspase-12蛋白表达。结果与假手术组相比,UUO模型组可见:1)肾小管扩张和肾间质纤维化程度随输尿管梗阻时间延长而渐进性加重;2)GRP78、CHOP、caspase-3及caspase-12蛋白表达在术后3 d均有上调,随着梗阻时间延长,上述蛋白表达更显著(P<0.01)。结论内质网应激相关标志性蛋白在UUO大鼠肾间质纤维化的早期即存在表达上调,可能促进了肾间质纤维化不断进展。展开更多
Huanglian Jiedu Decoction(HLJDD)is a quintessential prescription renowned for its heat-clearing and detoxifying properties.It is primarily prescribed to counteract the syndrome characterized by the excessive heat of t...Huanglian Jiedu Decoction(HLJDD)is a quintessential prescription renowned for its heat-clearing and detoxifying properties.It is primarily prescribed to counteract the syndrome characterized by the excessive heat of the Sanjiao fire.Notably,the hyperactivity of liver fire is frequently linked with hypertension,where wind fire and wind toxicity emerge as pivotal pathogenic factors.This study aimed to investigate the impact of HLJDD on the endoplasmic reticulum in spontaneously hypertensive rats(SHR),further delving into the interplay between endoplasmic reticulum stress(ERS)and myocardial remodeling and damage.Fifty SHR rats were stratified randomly into five cohorts:model,low-dose HLJDD,medium-dose HLJDD,high-dose HLJDD,and captopril groups.For comparison,a set of Wistar-Kyoto(WKY)rats served as the baseline control group,with each group comprising 10 rats.While the model and control groups received equivalent volumes of normal saline via gavage,the other groups were administered the respective drug dosages through the same route daily for a span of 6 weeks.Upon the experiment’s conclusion,metrics such as the heart mass index(HWI)and left ventricular mass index(LVWI)were assessed.Cardiac tissue anomalies were identified using H&E staining,while ERS-related protein and mRNA expression levels were ascertained via Western blotting analysis and qPCR.Moreover,TUNEL staining was employed to detect cardiomyocyte apoptosis.The findings indicated that increasing HLJDD concentrations corresponded with escalated HWI and LVWI in rat hearts(P<0.05).There was a marked enhancement in myocardial structural integrity,accompanied by a notable reduction in collagen fibers.The mRNA and protein expressions of myocardial inositol-dependent enzyme 1α(IRE1α),X-box binding protein 1(XBP1),glycoregulatory protein 78(GRP78),and CCAAT enhancer binding protein homologous protein(CHOP)in the medium and high-dose groups saw significant declines(P<0.05).These effects mirrored those observed in the captopril group.The study underscored HLJDD’s efficacy in mitigating myocardial tissue damage in SHR.This therapeutic effect was potentially attributed to the downregulation of IRE1α,XBP1,GRP78,and CHOP,curbing excessive ERS,diminishing cardiomyocyte apoptosis,and thereby conferring cardioprotection.展开更多
文摘目的探究内质网应激(ERS)相关凋亡通路在单侧输尿管梗阻(UUO)大鼠肾间质纤维化中的作用机制。方法将大鼠随机分为假手术组及UUO模型组,UUO组行左侧输尿管结扎术,于术后3、7和14 d HE和Masson染色观察肾脏病理变化;眼底静脉丛取血,分离血清测定血尿素氮及肌酐;Western blot检测葡萄糖调节蛋白78(GRP78)、内质网源性转录因子(CHOP)、凋亡相关蛋白半胱氨酸天门冬氨酸蛋白酶3(caspase-3)及caspase-12蛋白表达。结果与假手术组相比,UUO模型组可见:1)肾小管扩张和肾间质纤维化程度随输尿管梗阻时间延长而渐进性加重;2)GRP78、CHOP、caspase-3及caspase-12蛋白表达在术后3 d均有上调,随着梗阻时间延长,上述蛋白表达更显著(P<0.01)。结论内质网应激相关标志性蛋白在UUO大鼠肾间质纤维化的早期即存在表达上调,可能促进了肾间质纤维化不断进展。
基金The National Natural Science Foundation of China(Grant No.81860831)the Shandong Provincial Key Project of Traditional Chinese Medicine Science and Technology(Grant Nos.M-2023170,M-2022233).
文摘Huanglian Jiedu Decoction(HLJDD)is a quintessential prescription renowned for its heat-clearing and detoxifying properties.It is primarily prescribed to counteract the syndrome characterized by the excessive heat of the Sanjiao fire.Notably,the hyperactivity of liver fire is frequently linked with hypertension,where wind fire and wind toxicity emerge as pivotal pathogenic factors.This study aimed to investigate the impact of HLJDD on the endoplasmic reticulum in spontaneously hypertensive rats(SHR),further delving into the interplay between endoplasmic reticulum stress(ERS)and myocardial remodeling and damage.Fifty SHR rats were stratified randomly into five cohorts:model,low-dose HLJDD,medium-dose HLJDD,high-dose HLJDD,and captopril groups.For comparison,a set of Wistar-Kyoto(WKY)rats served as the baseline control group,with each group comprising 10 rats.While the model and control groups received equivalent volumes of normal saline via gavage,the other groups were administered the respective drug dosages through the same route daily for a span of 6 weeks.Upon the experiment’s conclusion,metrics such as the heart mass index(HWI)and left ventricular mass index(LVWI)were assessed.Cardiac tissue anomalies were identified using H&E staining,while ERS-related protein and mRNA expression levels were ascertained via Western blotting analysis and qPCR.Moreover,TUNEL staining was employed to detect cardiomyocyte apoptosis.The findings indicated that increasing HLJDD concentrations corresponded with escalated HWI and LVWI in rat hearts(P<0.05).There was a marked enhancement in myocardial structural integrity,accompanied by a notable reduction in collagen fibers.The mRNA and protein expressions of myocardial inositol-dependent enzyme 1α(IRE1α),X-box binding protein 1(XBP1),glycoregulatory protein 78(GRP78),and CCAAT enhancer binding protein homologous protein(CHOP)in the medium and high-dose groups saw significant declines(P<0.05).These effects mirrored those observed in the captopril group.The study underscored HLJDD’s efficacy in mitigating myocardial tissue damage in SHR.This therapeutic effect was potentially attributed to the downregulation of IRE1α,XBP1,GRP78,and CHOP,curbing excessive ERS,diminishing cardiomyocyte apoptosis,and thereby conferring cardioprotection.
