Hebei University of Chinese Medicine, Shijiazhuang, China.
Hebei University of Chinese Medicine, Shijiazhuang, China.
J Ethnopharmacol. 2022 Jan 30;283:114635. doi: 10.1016/j.jep.2021.114635. Epub 2021 Oct 12.
Huazhuojiedu decoction, a Chinese herbal preparation, has been proven to be clinically effective in treating precancerous lesions in gastric cancer (PLGC). This formula is optimized from a classic formula called "Ganluxiaodu Dan." Although some experiments have shown that Huazhuojiedu decoction is effective against PLGC, the mechanism remains unclear.
To investigate the treatment of PLGC with Huazhuojiedu decoction from the perspective of lncRNA in vitro and in vivo.
A PLGC rat model was prepared and randomly divided into a Huazhuojiedu decoction group (HG), a vitacoenzyme group (VG), a model group (MG), and a normal group (CG). Each group was given a corresponding concentration of medicine and distilled water for 10 weeks. The pathological changes in the gastric mucosa were observed by hematoxylin-eosin staining (HE). High-throughput sequencing was performed to detect the differentially expressed lncRNAs in the HG, MG, and CG. Quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR) was used to verify differentially expressed lncRNAs, and rat-human homology information was obtained from the University of California, Santa Cruz (UCSC) Genome Database. Human gastric mucosal epithelial cells (GES-1) were used to prepare precancerous lesions of gastric cancer cells (MC). A Huazhuojiedu decoction drug-containing serum was prepared to treat the MC cells. The effects of the Huazhuojiedu decoction and the lncRNA ENST00000517368 (lnc 517368) knockdown or overexpression on PLGC cell proliferation and apoptosis were evaluated in vitro using CCK-8, flow cytometry, and RT-qPCR.
The HE results showed that gastric mucosal pathology was significantly improved in the HG. High-throughput sequencing results showed that compared with the CG, 91 lncRNAs upregulated in the MG were restored and downregulated in the HG (P < 0.05), and 115 lncRNAs downregulated in the MG were restored and upregulated in the HG (P < 0.05). The results of RT-qPCR were consistent with the sequencing results. The differentially expressed genomic rat lncRNA ENSRNOT00000079699 is homologous to human lnc 517368. In cell experiments, high expression of lnc 517368 promoted proliferation and reduced apoptosis in PLGC cells, while the Huazhuojiedu decoction reduced the expression of lnc 517368 and improved cell morphology.
Huazhuojiedu decoction inhibited cell proliferation and promoted apoptosis in PLGC cells, and its effect may be partially dependent on the downregulation of lnc 517368.
化浊解毒汤是一种中药制剂，已被证明在治疗胃癌前病变（PLGC）方面具有临床疗效。该配方是从一种名为“肝瘤消毒丹”的经典配方优化而来。尽管一些实验表明化浊解毒汤对 PLGC 有效，但作用机制尚不清楚。
从长链非编码 RNA（lncRNA）的角度探讨化浊解毒汤治疗 PLGC 的作用机制。
建立 PLGC 大鼠模型，随机分为化浊解毒汤组（HG）、维酶素组（VG）、模型组（MG）和正常组（CG），每组给予相应浓度的药物和蒸馏水，连续给药 10 周。采用苏木精-伊红（HE）染色观察胃黏膜的病理变化。对 HG、MG 和 CG 中的差异表达 lncRNAs 进行高通量测序。采用实时荧光定量逆转录聚合酶链反应（RT-qPCR）验证差异表达的 lncRNAs，并从加利福尼亚大学圣克鲁兹分校（UCSC）基因组数据库获取人源胃黏膜上皮细胞（GES-1）的同源信息。制备化浊解毒汤含药血清，处理人胃癌前病变细胞（MC）。采用 CCK-8、流式细胞术和 RT-qPCR 评价化浊解毒汤和 lncRNA ENST00000517368（lnc517368）敲低或过表达对 PLGC 细胞增殖和凋亡的影响。
HE 结果显示，HG 组胃黏膜病理明显改善。高通量测序结果显示，与 CG 相比，MG 中 91 个上调的 lncRNAs 在 HG 中恢复为下调，115 个下调的 lncRNAs在 HG 中恢复为上调（P<0.05）。RT-qPCR 结果与测序结果一致。差异表达的基因组大鼠 lncRNA ENSRNOT00000079699 与人类 lnc517368 同源。细胞实验结果显示，lnc517368 高表达促进 PLGC 细胞增殖，减少细胞凋亡，而化浊解毒汤降低 lnc517368 的表达，改善细胞形态。
化浊解毒汤抑制 PLGC 细胞增殖，促进细胞凋亡，其作用可能部分依赖于下调 lnc517368。
