背景:慢性萎缩性胃炎(CAG)是一种慢性消化系统疾病。现代研究表明,CAG的进展与氧化应激诱导的胃粘膜DNA损伤和细胞凋亡的发生密切相关。此外,研究表明,Costunolide(COS),在木香中发现的主要活性化合物,一种传统的草药,表现出抗氧化性能。然而,COS治疗CAG的潜力及其分子靶点尚未确定.
目的:本研究的目的是探讨COS对N-甲基-N'-硝基-N-亚硝基胍(MNNG)诱导的CAG的胃粘膜保护作用及其机制。
方法:首先,建立MNNG诱导的大鼠体内CAG模型。通过胃的宏观检查和H&E染色来检测CAG的发生。此外,我们评估了氧化应激,DNA损伤,和使用生化检测的细胞凋亡,蛋白质印迹,免疫组织化学和免疫荧光。然后,建立了一个体外模型来诱导MNNG诱导的GES-1细胞损伤,并通过Hoechst33,342染色和流式细胞术确定细胞损伤的发生。最后,通过分子对接确定了COS治疗CAG的关键靶点,细胞热转移测定(CETSA),和抑制剂ML385。
结果:体内研究表明COS促进胃组织中Nrf2的表达。这导致SOD的表达增加,GSH,HO-1,同时降低MDA的产生。此外,COS通过抑制胃组织中γH2AX和PARP1的表达来抑制DNA损伤和凋亡。体外研究表明,COS可有效逆转MNNG诱导的GES-1细胞凋亡。此外,COS与Nrf2相互作用促进其表达。此外,SOD的表达水平,GSH,HO-1被增强,而ROS和MDA的生成减少。
结论:我们的结果表明,COS通过促进Nrf2的表达,抑制氧化应激和DNA损伤对CAG具有治疗作用。因此,COS有潜力为CAG的医治供给新药。
BACKGROUND: Chronic atrophic gastritis (CAG) is a chronic digestive disease. Modern research has revealed substantial evidence indicating that the progression of CAG is closely linked to the occurrence of oxidative stress-induced DNA damage and apoptosis in the gastric mucosa. Additionally, research has indicated that Costunolide (COS), the primary active compound found in Aucklandiae Radix, a traditional herb, exhibits antioxidant properties. Nevertheless, the therapeutic potential of COS in treating CAG and its molecular targets have not yet been determined.
OBJECTIVE: The objective of this research was to explore the potential gastric mucosal protective effects and mechanisms of COS against N-Methyl-N´-nitro-N-nitrosoguanidine (MNNG)-induced CAG.
METHODS: Firstly, the MNNG-induced rat CAG model was established in vivo. Occurrence of CAG was detected through macroscopic examination of the stomachs and H&E staining. Additionally, we assessed oxidative stress, DNA damage, and apoptosis using biochemical detection, Western blot, immunohistochemistry and immunofluorescence. Then, an in vitro model was developed to induce MNNG-induced damage in GES-1 cells, and the occurrence of cell damage was determined by Hoechst 33,342 staining and flow cytometry. Finally, the key targets of COS for the treatment of CAG were identified through molecular docking, cellular thermal shift assay (CETSA), and inhibitor ML385.
RESULTS: In vivo studies demonstrated that COS promotes the expression of Nrf2 in gastric tissues. This led to an increased expression of SOD, GSH, HO-1, while reducing the production of MDA. Furthermore, COS inhibited DNA damage and apoptosis by suppressing the expression of γH2AX and PARP1 in gastric tissues. In vitro studies showed that COS effectively reversed apoptosis induced by MNNG in GES-1 cells. Additionally, COS interacted with Nrf2 to promote its expression. Furthermore, the expression levels of SOD, GSH, and HO-1 were augmented, while the generation of ROS and MDA was diminished.
CONCLUSIONS: Our results indicate that COS exhibits therapeutic effects on CAG through the promotion of Nrf2 expression and inhibition of oxidative stress and DNA damage. Therefore, COS has the potential to provide new drugs for the treatment of CAG.