Abstract：
BACKGROUND: Achyranthes aspera, a widely recognized medicinal plant, is used in various cultures for treating different ailments, including renal dysfunction; however, there is a lack of comprehensive understanding of its protective effects and the underlying signaling networks involved.
OBJECTIVE: This study aimed to investigate the molecular mechanisms of the action of A. aspera by employing an integrative approach including functional and tissue imaging as well as comprehensive genomics analysis.
METHODS: Cisplatin-induced nephrotoxicity is a well-established animal model for acute kidney injury (AKI). In this study, we investigated the protective effects and underlying mechanisms of the action of A. aspera water-soluble extract (AAW) on a murine model of cisplatin-induced AKI. The evaluation includes measurements of blood urea nitrogen (BUN) and serum creatinine (SCr) levels, histology examination, and transcriptome analysis using RNA sequencing.
RESULTS: In male ICR mice, oral administration of AAW at doses of 0.5-1.0 g/kg significantly reduced cisplatin-induced nephrotoxicity. This effect included the amelioration of tubular injury, renal fibrosis, and the lowering of BUN and SCr levels. AAW also effectively decreased oxidative markers, such as malondialdehyde (MDA) and nitrotyrosine (NT), along with inflammation markers, including COX-2, iNOS, NLRP3, and pP65NFκB. Moreover, AAW administration induced a dose-dependent increase in the expression of two protective factors, Nrf2 and BcL2, and suppressed apoptosis, as evidenced by reduced levels of truncated caspase 3 (t-Casp3). To explore the underlying molecular mechanisms and signaling networks, next-generation sequencing (NGS) analysis was employed. The results revealed that AAW mitigated apoptosis, necroptosis, and PANoptosis pathways by inhibiting inflammation signaling pathways, such as the TNFα-, NFκB-, NETs-, and leukocyte transendothelial migration pathways. Additionally, AAW was found to enhance protective signaling pathways, including the cGMP/PKG-, cAMP-, AMPK-, and mTOR-dependent activation of autophagy and mitophagy pathways. The primary bioactive compound found in AAW was identified as 20-hydroxyecdysone (0.36%).
CONCLUSIONS: Our study demonstrates that AAW reduces cisplatin-induced nephrotoxicity. The protective effects of AAW are attributed to its modulation of multiple molecular signaling networks. Specifically, AAW downregulates genes and signaling pathways associated with oxidative stress and endoplasmic reticulum (ER) stress, inflammation, and PANoptosis. Simultaneously, it upregulates genes and signaling pathways associated with cell survival, including autophagy and mitophagy pathways.
OBJECTIVE: This study aimed to investigate the molecular mechanisms of the action of A. aspera by employing an integrative approach including functional and tissue imaging as well as comprehensive genomics analysis.
METHODS: Cisplatin-induced nephrotoxicity is a well-established animal model for acute kidney injury (AKI). In this study, we investigated the protective effects and underlying mechanisms of the action of A. aspera water-soluble extract (AAW) on a murine model of cisplatin-induced AKI. The evaluation includes measurements of blood urea nitrogen (BUN) and serum creatinine (SCr) levels, histology examination, and transcriptome analysis using RNA sequencing.
RESULTS: In male ICR mice, oral administration of AAW at doses of 0.5-1.0 g/kg significantly reduced cisplatin-induced nephrotoxicity. This effect included the amelioration of tubular injury, renal fibrosis, and the lowering of BUN and SCr levels. AAW also effectively decreased oxidative markers, such as malondialdehyde (MDA) and nitrotyrosine (NT), along with inflammation markers, including COX-2, iNOS, NLRP3, and pP65NFκB. Moreover, AAW administration induced a dose-dependent increase in the expression of two protective factors, Nrf2 and BcL2, and suppressed apoptosis, as evidenced by reduced levels of truncated caspase 3 (t-Casp3). To explore the underlying molecular mechanisms and signaling networks, next-generation sequencing (NGS) analysis was employed. The results revealed that AAW mitigated apoptosis, necroptosis, and PANoptosis pathways by inhibiting inflammation signaling pathways, such as the TNFα-, NFκB-, NETs-, and leukocyte transendothelial migration pathways. Additionally, AAW was found to enhance protective signaling pathways, including the cGMP/PKG-, cAMP-, AMPK-, and mTOR-dependent activation of autophagy and mitophagy pathways. The primary bioactive compound found in AAW was identified as 20-hydroxyecdysone (0.36%).
CONCLUSIONS: Our study demonstrates that AAW reduces cisplatin-induced nephrotoxicity. The protective effects of AAW are attributed to its modulation of multiple molecular signaling networks. Specifically, AAW downregulates genes and signaling pathways associated with oxidative stress and endoplasmic reticulum (ER) stress, inflammation, and PANoptosis. Simultaneously, it upregulates genes and signaling pathways associated with cell survival, including autophagy and mitophagy pathways.
摘要：
背景：牛膝，一种广受认可的药用植物，在各种文化中用于治疗不同的疾病，包括肾功能不全；然而，对其保护作用和所涉及的潜在信令网络缺乏全面的了解。
目的：本研究旨在通过采用包括功能和组织成像以及全面基因组学分析在内的综合方法来研究A.aspera作用的分子机制。
方法：顺铂诱导的肾毒性是一种公认的急性肾损伤（AKI）动物模型。在这项研究中,我们研究了Aspera水溶性提取物（AAW）对顺铂诱导的AKI小鼠模型的保护作用和潜在机制。评估包括测量血尿素氮（BUN）和血清肌酐（SCr）水平，组织学检查,和使用RNA测序的转录组分析。
结果：在雄性ICR小鼠中，口服0.5-1.0g/kg剂量的AAW可显着降低顺铂诱导的肾毒性。这种效果包括改善肾小管损伤，肾纤维化,以及BUN和SCr水平的降低。AAW还有效降低了氧化标志物，如丙二醛（MDA）和硝基酪氨酸（NT），以及炎症标志物，包括COX-2，iNOS，NLRP3和pP65NFκB。此外，AAW给药诱导两个保护因子表达的剂量依赖性增加，Nrf2和BcL2，抑制细胞凋亡，正如截短的半胱天冬酶3(t-Casp3)水平降低所证明的。探索潜在的分子机制和信号网络，采用下一代测序(NGS)分析.结果表明，AAW减轻了细胞凋亡，坏死,通过抑制炎症信号通路，例如TNFα-，NFκB-,网络-,和白细胞跨内皮迁移途径。此外,AAW被发现可以增强保护性信号通路，包括cGMP/PKG-，cAMP-,AMPK-,和mTOR依赖性自噬和线粒体自噬途径的激活。在AAW中发现的主要生物活性化合物被鉴定为20-羟基cedysone(0.36%)。
结论：我们的研究表明，AAW降低了顺铂诱导的肾毒性。AAW的保护作用归因于其对多个分子信号网络的调节。具体来说,AAW下调与氧化应激和内质网（ER）应激相关的基因和信号通路，炎症,和全景。同时,它上调与细胞存活相关的基因和信号通路，包括自噬和线粒体自噬途径。
目的：本研究旨在通过采用包括功能和组织成像以及全面基因组学分析在内的综合方法来研究A.aspera作用的分子机制。
方法：顺铂诱导的肾毒性是一种公认的急性肾损伤（AKI）动物模型。在这项研究中,我们研究了Aspera水溶性提取物（AAW）对顺铂诱导的AKI小鼠模型的保护作用和潜在机制。评估包括测量血尿素氮（BUN）和血清肌酐（SCr）水平，组织学检查,和使用RNA测序的转录组分析。
结果：在雄性ICR小鼠中，口服0.5-1.0g/kg剂量的AAW可显着降低顺铂诱导的肾毒性。这种效果包括改善肾小管损伤，肾纤维化,以及BUN和SCr水平的降低。AAW还有效降低了氧化标志物，如丙二醛（MDA）和硝基酪氨酸（NT），以及炎症标志物，包括COX-2，iNOS，NLRP3和pP65NFκB。此外，AAW给药诱导两个保护因子表达的剂量依赖性增加，Nrf2和BcL2，抑制细胞凋亡，正如截短的半胱天冬酶3(t-Casp3)水平降低所证明的。探索潜在的分子机制和信号网络，采用下一代测序(NGS)分析.结果表明，AAW减轻了细胞凋亡，坏死,通过抑制炎症信号通路，例如TNFα-，NFκB-,网络-,和白细胞跨内皮迁移途径。此外,AAW被发现可以增强保护性信号通路，包括cGMP/PKG-，cAMP-,AMPK-,和mTOR依赖性自噬和线粒体自噬途径的激活。在AAW中发现的主要生物活性化合物被鉴定为20-羟基cedysone(0.36%)。
结论：我们的研究表明，AAW降低了顺铂诱导的肾毒性。AAW的保护作用归因于其对多个分子信号网络的调节。具体来说,AAW下调与氧化应激和内质网（ER）应激相关的基因和信号通路，炎症,和全景。同时,它上调与细胞存活相关的基因和信号通路，包括自噬和线粒体自噬途径。
