目的 探討刺五加注射液(CWJI)對(duì)慢性萎縮性胃炎(CAG)癌前病變模型大鼠的作用及機(jī)制。方法 采用 N-甲基-N'-硝基-N-亞硝基 胍(MNNG)復(fù)合法構(gòu)建CAG癌前病變大鼠模型,模型大鼠隨機(jī)分為模型組、胃復(fù)春膠囊(400 g·kg^-1,ig給藥)組和 CWJI 低、中、高劑量(15、30、60 mg·kg^-1,臨床等效劑量的 0.5、1、2 倍,ip 給藥)組,每組 12 只,另取 12 只大鼠設(shè)為對(duì)照組,對(duì)照組和模型組 ip 給予等體積 0.9% 氯化鈉溶液,每天給藥 1 次,共給藥 12 周。HE 染色后光學(xué)顯微鏡觀察胃黏膜病理變化,醋酸鈾和枸櫞酸鉛雙染色后電子顯微鏡觀察胃黏膜上皮細(xì)胞超微結(jié)構(gòu)變化;ELISA 法檢測(cè)血清胃蛋白酶原 I(PG I)、PG II、胃泌素(GAS)、白細(xì)胞介素(IL)^-1β、IL-6、腫瘤壞死因子-α(TNF-α)水平;流式細(xì)胞術(shù)檢測(cè)外周血中自然殺傷(NK)細(xì)胞和 T 淋巴細(xì)胞亞群比例;實(shí)時(shí)熒光定量 PCR(qRT-PCR)法、Western blotting 法檢測(cè)胃黏膜缺氧誘導(dǎo)因子1α(HIF^-1α)、血管內(nèi)皮生長(zhǎng)因子 A(VEGFA)、血管內(nèi)皮生長(zhǎng)因子受體 2(VEGFR2)、核因子-κB(NF-κB)mRNA 和蛋白表達(dá)。結(jié)果 與模型組比較,胃復(fù)春膠囊組和 CWJI 中、高劑量組大鼠胃黏膜變薄、腺泡萎縮、新生血管增生、炎性細(xì)胞浸潤(rùn)以及腸上皮化生、上皮內(nèi)瘤變等病理改變明顯改善,胃黏膜上皮細(xì)胞胞核大小不等、核分裂相增多、染色質(zhì)結(jié)構(gòu)異常等超微結(jié)構(gòu)改變明顯減輕;血清 PG I、PG II、GAS 水平和 PG I/PG II值顯著提高,IL^-1β、IL-6、TNF-α 水平顯著降低(P<0.05);外周血 NK 細(xì)胞、CD³⁺ T 淋巴細(xì)胞、CD⁴⁺ T淋巴細(xì)胞比例和 CD⁴⁺/CD⁸⁺值顯著升高(P<0.05),CD8+ T 淋巴細(xì)胞比例顯著降低(P<0.05);胃黏膜HIF^-1α、VEGFA、VEGFR2、NF-κB mRNA和蛋白表達(dá)量顯著降低(P<0.05)。結(jié)論 CWJI 可阻斷或逆轉(zhuǎn)大鼠CAG癌前病變,其機(jī)制可能與提高細(xì)胞免疫功能以及調(diào)控 HIF^-1α/VEGFA/NF-κB信號(hào)通路,抑制血管新生和炎癥反應(yīng)有關(guān)。

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

河北省邯鄲市科學(xué)技術(shù)研究與發(fā)展計(jì)劃項(xiàng)目(21422083020)