目的 探讨刺五加注射液(CWJI)对慢性萎缩性胃炎(CAG)癌前病变模型大鼠的作用及机制。方法 采用 N-甲基-N'-硝基-N-亚硝基 胍(MNNG)复合法构建CAG癌前病变大鼠模型,模型大鼠随机分为模型组、胃复春胶囊(400 g·kg^-1,ig给药)组和 CWJI 低、中、高剂量(15、30、60 mg·kg^-1,临床等效剂量的 0.5、1、2 倍,ip 给药)组,每组 12 只,另取 12 只大鼠设为对照组,对照组和模型组 ip 给予等体积 0.9% 氯化钠溶液,每天给药 1 次,共给药 12 周。HE 染色后光学显微镜观察胃黏膜病理变化,醋酸铀和枸橼酸铅双染色后电子显微镜观察胃黏膜上皮细胞超微结构变化;ELISA 法检测血清胃蛋白酶原 I(PG I)、PG II、胃泌素(GAS)、白细胞介素(IL)^-1β、IL-6、肿瘤坏死因子-α(TNF-α)水平;流式细胞术检测外周血中自然杀伤(NK)细胞和 T 淋巴细胞亚群比例;实时荧光定量 PCR(qRT-PCR)法、Western blotting 法检测胃黏膜缺氧诱导因子1α(HIF^-1α)、血管内皮生长因子 A(VEGFA)、血管内皮生长因子受体 2(VEGFR2)、核因子-κB(NF-κB)mRNA 和蛋白表达。结果 与模型组比较,胃复春胶囊组和 CWJI 中、高剂量组大鼠胃黏膜变薄、腺泡萎缩、新生血管增生、炎性细胞浸润以及肠上皮化生、上皮内瘤变等病理改变明显改善,胃黏膜上皮细胞胞核大小不等、核分裂相增多、染色质结构异常等超微结构改变明显减轻;血清 PG I、PG II、GAS 水平和 PG I/PG II值显著提高,IL^-1β、IL-6、TNF-α 水平显著降低(P<0.05);外周血 NK 细胞、CD³⁺ T 淋巴细胞、CD⁴⁺ T淋巴细胞比例和 CD⁴⁺/CD⁸⁺值显著升高(P<0.05),CD8+ T 淋巴细胞比例显著降低(P<0.05);胃黏膜HIF^-1α、VEGFA、VEGFR2、NF-κB mRNA和蛋白表达量显著降低(P<0.05)。结论 CWJI 可阻断或逆转大鼠CAG癌前病变,其机制可能与提高细胞免疫功能以及调控 HIF^-1α/VEGFA/NF-κB信号通路,抑制血管新生和炎症反应有关。

Objective To investigate the effect of Ciwujia Injection (CWJI) on precancerous lesions model rats of chronic atrophic gastritis (CAG) and explore its possible mechanism. Methods The CAG precancerous lesion model rats were established by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) compound method. The model rats were randomly divided into model group, Weifuchun Capsule (400 g·kg^-1, ig administration) group, and CWJI low, medium and high dose (15, 30, 60 mg·kg^-1, respectively, 0.5, 1, 2 times of the clinical equivalent dose, ip administration) group, another 12 rats in each group, 12 rats were selected as control group, rats in control group and model group were ip given the same volume of 0.9% sodium chloride solution, once a day, a total of 12 weeks of administration. The pathological changes of gastric mucosa was observed by optical microscope after HE staining, the ultrastructural changes of gastric mucosa epithelial cells was observed by electron microscope after double staining with uranyl acetate and lead citrate. The contents of pepsinogen I (PG I), PG II, gastric (GAS), interleukin (IL)-1β, IL-6, tumor necrosis factor α (TNF-α) were detected by ELISA method. The proportion of natural killer (NK) cells and T lymphocyte subsets in peripheral blood were detected by flow cytometry. The mRNA and protein expressions of hypoxia-inducible factor 1α (HIF-1α), vascular endothelial growth factor A (VEGFA), vascular endothelial growth factor receptor 2 (VEGFR2), nuclear factor κB (NF-κB) in gastric mucosa were detected by real-time quantitative fluorescence PCR (qRT-PCR) and Western blotting method. Results Compared with model group, the pathological changes of gastric mucosa such as gastric mucosal became thinner, acinar atrophy, neovascularization, inflammatory cell infiltration, intestinal metaplasia, intraepithelial neoplasia of the rats in Weifuchun Capsule group and CWJI medium, high dose groups were significantly improved. Compared with model group, the ultrastructural changes such as unequal size of nuclei, mitotic phase increased, chromatin structure abnormal of epithelial cells of the rats in Weifuchun Capsule group and CWJI medium, high dose groups were significantly alleviated. Compared with model group, the contents of PG I, PG II, GAS in serum and the ratio of PG I/PG II were significantly increased, the contents of IL-1β, IL-6, TNF-α in serum were significantly decreased (P<0.05), the proportion of NK cells, CD³⁺, CD⁴⁺ T lymphocytes in peripheral blood and the ratio of CD⁴⁺/CD⁸⁺ were significantly increased (P<0.05), while the proportion of CD⁸⁺ T lymphocyte was significantly decreased (P<0.05), the mRNA and protein expression of HIF-1α, VEGFA, VEGFR2, NF-κB in gastric mucosa were significantly decreased (P<0.05) in Weifuchun Capsule group and CWJI medium, high dose groups. Conclusion CWJI can block or revers CAG precancerous lesions in rats, which mechanism may be related to the improvement of cellular immune function and regulating HIF-1α/VEGFA/NF-κB signaling pathway, inhibiting angiogenesis and inflammation.

R285.5

河北省邯郸市科学技术研究与发展计划项目(21422083020)