基于IL-6/JAK2/STAT3通路探讨化瘀消痞汤对胃癌前病变大鼠胃黏膜损伤的作用及机制

Exploration of Huayu Xiaopi Decoction on Improving Gastric Mucosal Injury in Rats with Precancerous Lesions of Gastric Cancer Based on IL-6/JAK2/STAT3 Pathway

目的基于白细胞介素6(IL-6)/Janus激酶2(JAK2)/信号转导和转录激活因子3(STAT3)通路探讨化瘀消痞汤对胃癌前病变(PLGC)大鼠胃黏膜损伤的作用及机制。方法随机选取10只大鼠作为空白组,其余50只大鼠作为造模组,采用多因素综合造模。将模型复制成功的大鼠随机分为模型组、叶酸组(0.002 g·kg^(-1))及化瘀消痞汤低、中、高剂量组(6.2、12.4、24.8 g·kg^(-1))。灌胃给药,每日1次,连续干预3个月。观察大鼠一般状况并测定体质量、3 h进食量;采用HE染色法观察胃组织病理变化;ELISA法检测血清中IL-6、肿瘤坏死因子α(TNF-α)、IL-1β水平;免疫荧光法检测胃组织中B细胞淋巴瘤/白血病-xL(Bcl-xL)、血管内皮生长因子(VEGF)蛋白表达水平;WesternBlot法检测胃组织中IL-6、p-JAK2、p-STAT3、细胞周期素D1(CyclinD1)、原癌基因(c-Myc)蛋白表达水平。结果与空白组比较,模型组大鼠体质量、3 h进食量明显降低(P<0.05);胃黏膜明显变薄且不完整,腺体排列紊乱且数量减少,部分区域可见胃上皮细胞肠化的杯状细胞及炎性细胞浸润;血清IL-6、TNF-α、IL-1β水平明显升高(P<0.05);胃组织中Bcl-xL、VEGF、IL-6、p-JAK2、p-STAT3、CyclinD1、c-Myc蛋白表达明显上调(P<0.05)。与模型组比较,化瘀消痞汤中、高剂量组大鼠的体质量、3 h进食量明显升高(P<0.05);胃黏膜损伤有不同程度改善,腺体排列趋于整齐,胃上皮细胞肠化情况有所改善,炎性细胞较少;血清IL-6、TNF-α、IL-1β水平明显降低(P<0.05);胃组织中Bcl-xL、VEGF、IL-6、p-JAK2、p-STAT3、CyclinD1、c-Myc蛋白表达明显下调(P<0.05)。结论化瘀消痞汤能明显改善PLGC大鼠的胃黏膜病理损伤,可能与其抑制炎症反应及调控IL-6/JAK2/STAT3通路抑制细胞增殖有关。

Objective To explore the effect and mechanism of Huayu Xiaopi Decoction on gastric mucosal injury in rats with precancerous lesions of gastric cancer(PLGC)based on the interleukin-6(IL-6)/Janus kinase 2(JAK2)/signal transduction and transcription activating factor 3(STAT3)pathway.Methods Totally 10 SD rats were randomly selected as a blank group and the remaining 50 rats were used as a model group,which were replicated using a multi-factor comprehensive modeling method.After the PLGC model was successfully constructed,rats were randomly divided into model group,folic acid group(0.002 g·kg^(-1)),low-,medium-and high-dose of Huayu Xiaopi Decoction(6.2,12.4,24.8 g·kg^(-1))groups,and then the corresponding drug was orally administered once a day for three months.The general condition,the body weight and 3-hour food intake of rats were observed.The pathological changes of gastric tissues in each group of rats were observed using hematoxylin-eosin(HE)staining.Enzyme linked immunosorbent assay(ELISA)was used to detect the expression of serum IL-6,interleukin-1β(IL-1β),and tumor necrosis factor alpha(TNF-α)in each rat group.The expression of B-cell lymphoma/leukemia xL(Bcl-xL)and vascular endothelial growth factor(VEGF)in the gastric tissue of rats in each group was detected by fluorescence immunohistochemistry.Western Blot was used to test the protein expression of IL-6,p-JAK2,p-STAT3,Cyclin D1 and c-Myc in the gastric tissues of rats in each group.Results Compared with the blank group,the body weight and 3-hour food intake of rats in the model group were significantly reduced(P<0.05),the gastric mucosa became thinner and incomplete.Disordered glandular arrangement and reduced glandular number were found.Goblet cells of intestinal metaplasia in gastric epithelial cells and infiltration of inflammatory cells were visible in partial area.The levels of IL-6,IL-1βand TNF-αin the rat serum were also significantly up-regulated(P<0.05),and the protein expression of IL-6,p-JAK2,p-STAT3,CyclinD1,c-Myc,Bcl-xL and VEGF in gastric tissue was obviously up-regulated(P<0.05).Compared with the model group,the body weight and 3-hour food intake of rats in medium-and high-dose of Huayu Xiaopi Decoction groups increased(P<0.05),the gastric mucosal injury were improved with different degree.Glandular arrangement was in order,intestinal metaplasia of gastric epithelial cells was improved,as well as less inflammatory cells were found.The expression of IL-6,TNF-αand IL-1βin the serum of rats remarkably decreased(P<0.05),while the protein expression of Bcl-xL,VEGF,IL-6,p-JAK2,p-STAT3,CyclinD1 and c-Myc in gastric tissue obviously down-regulated(P<0.05).Conclusion Huayu Xiaopi Decoction can effectively improve pathological damage of gastric mucosa in PLGC rats.Its mechanism maybe related to inhibiting inflammatory reaction,and suppressing cell proliferation by regulating the IL-6/JAK2/STAT3 pathway.