Abstract：
Heat shock proteins (HSPs), which function as chaperones, are activated in response to various environmental stressors. In addition to their role in diverse aspects of protein production, HSPs protect against harmful protein-related stressors. Calycosin exhibits numerous beneficial properties. This study aims to explore the protective effects of calycosin in the heart under heat shock and determine its underlying mechanism. H9c2 cells, western blot, TUNEL staining, flow cytometry, and immunofluorescence staining were used. The time-dependent effects of heat shock analyzed using western blot revealed increased HSP expression for up to 2[Formula: see text]h, followed by protein degradation after 4[Formula: see text]h. Hence, a heat shock damage duration of 4[Formula: see text]h was chosen for subsequent investigations. Calycosin administered post-heat shock demonstrated dose-dependent recovery of cell viability. Under heat shock conditions, calycosin prevented the apoptosis of H9c2 cells by upregulating HSPs, suppressing p-JNK, enhancing Bcl-2 activation, and inhibiting cleaved caspase 3. Calycosin also inhibited Fas/FasL expression and activated cell survival markers (p-PI3K, p-ERK, p-Akt), indicating their cytoprotective properties through PI3K/Akt activation and JNK inhibition. TUNEL staining and flow cytometry confirmed that calycosin reduced apoptosis. Moreover, calycosin reversed the inhibitory effects of quercetin on HSF1 and Hsp70 expression, illustrating its role in enhancing Hsp70 expression through HSF1 activation during heat shock. Immunofluorescence staining demonstrated HSF1 translocation to the nucleus following calycosin treatment, emphasizing its cytoprotective effects. In conclusion, calycosin exhibits pronounced protective effects against heat shock-induced damages by modulating HSP expression and regulating key signaling pathways to promote cell survival in H9c2 cells.
摘要：
热休克蛋白(HSPs),作为监护人，被激活以响应各种环境压力。除了它们在蛋白质生产的各个方面的作用，HSPs可防止有害的蛋白质相关应激源。木脂素表现出许多有益的性质。本研究旨在探讨毛囊素在热休克心脏中的保护作用，并确定其潜在机制。H9c2细胞，westernblot,TUNEL染色,流式细胞术,和免疫荧光染色。使用蛋白质印迹分析的热休克的时间依赖性效应显示HSP表达增加高达2[公式：见文本]h，随后在4[公式：见文本]h后进行蛋白质降解。因此，选择热冲击损伤持续时间为4[公式：参见文本]h用于后续研究。在热休克后施用的花叶素证明了细胞活力的剂量依赖性恢复。在热冲击条件下,calycosin通过上调HSPs阻止H9c2细胞凋亡，抑制p-JNK，增强Bcl-2激活，并抑制裂解的胱天蛋白酶3.毛黄蛋白酶还抑制Fas/FasL表达和激活的细胞存活标志物（p-PI3K，p-ERK,p-Akt),通过PI3K/Akt激活和JNK抑制表明它们的细胞保护特性。TUNEL染色和流式细胞术证实calycosin减少细胞凋亡。此外，环毛素逆转槲皮素对HSF1和Hsp70表达的抑制作用,说明其在热休克期间通过HSF1激活增强Hsp70表达中的作用。免疫荧光染色显示HSF1易位到细胞核后，强调其细胞保护作用。总之,calycosin通过调节HSP表达和调节关键信号通路以促进H9c2细胞的细胞存活，对热休克诱导的损伤具有明显的保护作用。
