Abstract：
BACKGROUND: Throughout Chinese history, Hydrangea paniculata Siebold has been utilized as a traditional medicinal herb to treat a variety of ailments associated to inflammation. In a number of immune-mediated kidney disorders, total coumarins extracted from Hydrangea paniculata (HP) have demonstrated a renal protective effect.
OBJECTIVE: To investigate renal beneficial effect of HP on experimental Adriamycin nephropathy (AN), and further clarify whether reversing lipid metabolism abnormalities by HP contributes to its renoprotective effect and find out the underlying critical pathways.
METHODS: After establishment of rat AN model, HP was orally administrated for 6 weeks. Biochemical indicators related to kidney injury were determined. mRNAs sequencing using kidney tissues were performed to clarify the underlying mechanism. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis, western blot, molecular docking, and drug affinity responsive target stability (DARTS) assay was carried out to further explore and confirm pivotal molecular pathways and possible target by which HP and 7-hydroxylcoumarin (7-HC) played their renal protection effect via modulating lipid metabolism.
RESULTS: HP could significantly improve renal function, and restore renal tubular abnormal lipid metabolism and interstitial fibrosis in AN. In vitro study demonstrated that HP and its main metabolite 7-HC could reduce ADR-induced intracellular lipid deposition and fibrosis characteristics in renal tubular cells. Mechanically, HP and 7-HC can activate AMP-activated protein kinase (AMPK) via direct interaction, which contributes to its lipid metabolism modulation effect. Moreover, HP and 7-HC can inhibit fibrosis by inhibiting CCAAT/enhancer binding protein beta (C/EBPβ) expression in renal tubular cells. Normalization of lipid metabolism by HP and 7-HC further provided protection of mitochondrial structure integrity and inhibited the nuclear factor kappa-B (NF-κB) pathway. Long-term toxicity using beagle dogs proved the safety of HP after one-month administration.
CONCLUSIONS: Coumarin derivates from HP alleviate adriamycin-induced lipotoxicity and fibrosis in kidney through activating AMPK and inhibiting C/EBPβ.
OBJECTIVE: To investigate renal beneficial effect of HP on experimental Adriamycin nephropathy (AN), and further clarify whether reversing lipid metabolism abnormalities by HP contributes to its renoprotective effect and find out the underlying critical pathways.
METHODS: After establishment of rat AN model, HP was orally administrated for 6 weeks. Biochemical indicators related to kidney injury were determined. mRNAs sequencing using kidney tissues were performed to clarify the underlying mechanism. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis, western blot, molecular docking, and drug affinity responsive target stability (DARTS) assay was carried out to further explore and confirm pivotal molecular pathways and possible target by which HP and 7-hydroxylcoumarin (7-HC) played their renal protection effect via modulating lipid metabolism.
RESULTS: HP could significantly improve renal function, and restore renal tubular abnormal lipid metabolism and interstitial fibrosis in AN. In vitro study demonstrated that HP and its main metabolite 7-HC could reduce ADR-induced intracellular lipid deposition and fibrosis characteristics in renal tubular cells. Mechanically, HP and 7-HC can activate AMP-activated protein kinase (AMPK) via direct interaction, which contributes to its lipid metabolism modulation effect. Moreover, HP and 7-HC can inhibit fibrosis by inhibiting CCAAT/enhancer binding protein beta (C/EBPβ) expression in renal tubular cells. Normalization of lipid metabolism by HP and 7-HC further provided protection of mitochondrial structure integrity and inhibited the nuclear factor kappa-B (NF-κB) pathway. Long-term toxicity using beagle dogs proved the safety of HP after one-month administration.
CONCLUSIONS: Coumarin derivates from HP alleviate adriamycin-induced lipotoxicity and fibrosis in kidney through activating AMPK and inhibiting C/EBPβ.
摘要：
背景：纵观中国历史，绣球花已被用作传统药草来治疗与炎症相关的各种疾病。在许多免疫介导的肾脏疾病中，从绣球花（HP）中提取的总香豆素具有肾脏保护作用。
目的：研究HP对实验性阿霉素肾病（AN）的肾脏有益作用，并进一步阐明HP逆转脂质代谢异常是否有助于其肾脏保护作用，并找出潜在的关键途径。
方法：建立大鼠AN模型后,HP口服给药6周。测定与肾损伤相关的生化指标。使用肾组织进行mRNA测序以阐明潜在的机制。京都基因和基因组百科全书（KEGG）途径分析，westernblot,分子对接,并进行了药物亲和反应靶标稳定性（DARTS）测定，以进一步探索和确认HP和7-羟基香豆素（7-HC）通过调节脂质代谢发挥肾脏保护作用的关键分子途径和可能靶标。
结果：HP能明显改善肾功能，恢复肾小管异常脂质代谢和间质纤维化。体外研究表明，HP及其主要代谢产物7-HC可以减轻ADR诱导的肾小管细胞内脂质沉积和纤维化特征。机械上,HP和7-HC可以通过直接相互作用激活AMP激活的蛋白激酶（AMPK），这有助于其对脂质代谢的调节作用。此外，HP和7-HC可以通过抑制肾小管细胞中CCAAT/增强子结合蛋白β（C/EBPβ）的表达来抑制纤维化。HP和7-HC使脂代谢正常化进一步保护了线粒体结构的完整性，并抑制了核因子κB（NF-κB）途径。使用比格犬的长期毒性证明了HP在一个月给药后的安全性。
结论：从HP衍生的香豆素通过激活AMPK和抑制C/EBPβ减轻阿霉素诱导的肾脏脂毒性和纤维化。
目的：研究HP对实验性阿霉素肾病（AN）的肾脏有益作用，并进一步阐明HP逆转脂质代谢异常是否有助于其肾脏保护作用，并找出潜在的关键途径。
方法：建立大鼠AN模型后,HP口服给药6周。测定与肾损伤相关的生化指标。使用肾组织进行mRNA测序以阐明潜在的机制。京都基因和基因组百科全书（KEGG）途径分析，westernblot,分子对接,并进行了药物亲和反应靶标稳定性（DARTS）测定，以进一步探索和确认HP和7-羟基香豆素（7-HC）通过调节脂质代谢发挥肾脏保护作用的关键分子途径和可能靶标。
结果：HP能明显改善肾功能，恢复肾小管异常脂质代谢和间质纤维化。体外研究表明，HP及其主要代谢产物7-HC可以减轻ADR诱导的肾小管细胞内脂质沉积和纤维化特征。机械上,HP和7-HC可以通过直接相互作用激活AMP激活的蛋白激酶（AMPK），这有助于其对脂质代谢的调节作用。此外，HP和7-HC可以通过抑制肾小管细胞中CCAAT/增强子结合蛋白β（C/EBPβ）的表达来抑制纤维化。HP和7-HC使脂代谢正常化进一步保护了线粒体结构的完整性，并抑制了核因子κB（NF-κB）途径。使用比格犬的长期毒性证明了HP在一个月给药后的安全性。
结论：从HP衍生的香豆素通过激活AMPK和抑制C/EBPβ减轻阿霉素诱导的肾脏脂毒性和纤维化。
