Hebei University of Chinese Medicine; The First Affiliated Hospital of Hebei University of Chinese Medicine.
The First Affiliated Hospital of Hebei University of Chinese Medicine.
J Vis Exp. 2023 Jun 30(196). doi: 10.3791/65271.
This research aims to explore the therapeutic effect and potential mechanisms of Huazhuojiedu decoction (HZJD) for alleviating precancerous lesions of gastric cancer (PLGC) both in vivo and in vitro. HZJD is a traditional Chinese herbal formula consisting of 11 herbs. Sprague-Dawley (SD) rats were randomly divided into four subgroups: control group, model group, positive drug group, and HZJD group. Hematoxylin-eosin (H&E) staining, high iron diamine-alcian blue (HID-AB) staining, alcian blue-periodic acid Schiff (AB-PAS) staining, immunohistochemistry, immunofluorescence, RT-qPCR, and Western blot assays were performed after 10 weeks of HZJD treatment. In vitro, the cell counting kit-8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) assays were used to detect cell proliferation. RT-qPCR and Western blot assays were performed to evaluate mitophagy levels. The results indicated that HZJD could retard the pathological progression in PLGC rats and reduce PLGC cell proliferation. Treatment with HZJD significantly increased the mRNA and protein expression levels of Sirt3, Foxo3a, Parkin, and LC3 II/I, while decreasing the mRNA and protein expression levels of p62 and Tomm20. HZJD was found to have the ability to reverse the decline in mitophagy activity both in vivo and in vitro. In conclusion, the study assessed the impact of HZJD and provided evidence regarding its potential molecular mechanism.
本研究旨在探讨化浊解毒方（HZJD）对胃癌前病变（PLGC）的体内外治疗作用及潜在机制。HZJD 是一种由 11 种草药组成的中药方剂。SD 大鼠随机分为 4 组：对照组、模型组、阳性药物组和 HZJD 组。经过 10 周的 HZJD 治疗后，进行苏木精-伊红（H&E）染色、高铁二胺-阿尔辛蓝（HID-AB）染色、阿尔辛蓝-过碘酸希夫（AB-PAS）染色、免疫组织化学、免疫荧光、RT-qPCR 和 Western blot 检测。在体外，使用细胞计数试剂盒-8（CCK-8）和 5-乙炔基-2'-脱氧尿苷（EdU）检测细胞增殖。进行 RT-qPCR 和 Western blot 检测以评估自噬水平。结果表明，HZJD 可延缓 PLGC 大鼠的病理进展，降低 PLGC 细胞增殖。HZJD 治疗可显著增加 Sirt3、Foxo3a、Parkin 和 LC3 II/I 的 mRNA 和蛋白表达水平，同时降低 p62 和 Tomm20 的 mRNA 和蛋白表达水平。HZJD 被发现具有在体内外逆转自噬活性下降的能力。综上所述，本研究评估了 HZJD 的影响，并提供了其潜在分子机制的证据。
