基于转录组学探讨参芪抑瘤方干预胃癌癌前病变模型大鼠分子机制

Exploration on the Mechanism of Shenqi Yiliu Prescription in a Rat Model of Precancerous Lesions of Gastric Carcinoma Based on Transcriptomics

目的 基于转录组学探讨参芪抑瘤方对胃癌前病变(PLGC)模型大鼠的干预机制。方法 采用复合因素造模法构建PLGC大鼠模型。将大鼠随机分为空白组、模型组、叶酸组(0.002 g/kg)和参芪抑瘤方高、中、低剂量组(39.6、19.8、9.9 g/kg),每组10只,分别予相应溶液灌胃,连续90 d。观察大鼠一般状况,HE染色观察胃黏膜形态,免疫荧光染色检测胃组织增殖细胞核抗原(PCNA)蛋白表达,转录组学筛选差异表达mRNA并富集差异表达通路,ELISA检测胃组织Bcl-xL、C-myc、周期蛋白D1(Cyclin D1)含量,RT-qPCR检测胃组织Bcl-xL、C-myc、Cyclin D1 mRNA表达,Western blot检测胃组织白细胞介素-6(IL-6)、Janus酪氨酸激酶2(JAK2)、信号传导与转录激活因子3(STAT3)、p-JAK2、p-STAT3蛋白表达。结果 与空白组比较,模型组大鼠体质量降低(P<0.05),胃黏膜结构紊乱,胃组织PCNA蛋白表达升高(P<0.05),胃组织Bcl-xL、C-myc、Cyclin D1含量及mRNA表达升高(P<0.05),IL-6、JAK2、STAT3、p-JAK2、p-STAT3蛋白表达升高(P<0.05);与模型组比较,参芪抑瘤方高、中剂量组大鼠体质量不同程度升高(P<0.05),各给药组大鼠胃黏膜异常形态均有不同程度改善,参芪抑瘤方各剂量组PCNA蛋白表达降低(P<0.05);转录组测序结果显示,JAK-STAT信号通路在空白组与模型组、模型组与参芪抑瘤方高剂量组中均有显著差异;参芪抑瘤方高、中剂量组大鼠胃组织Bcl-xL、C-myc、Cyclin D1含量及m RNA表达降低(P<0.05),IL-6、JAK2、STAT3、p-JAK2、p-STAT3蛋白表达降低(P<0.05)。结论 参芪抑瘤方能改善PLGC模型大鼠胃黏膜异常形态,其机制与调控IL-6/JAK2/STAT3信号通路从而抑制细胞增殖有关。

Objective To investigate the intervention mechanism of Shenqi Yiliu Prescription in a rat model of precancerous lesions of gastric carcinoma(PLGC)based on transcriptomics.Methods A PLGC rat model was constructed using composite factor modeling method.Rats were randomly divided into blank group,model group,folic acid group(0.002 g/kg),and Shenqi Yiliu Prescription high-,medium-and low-dosage groups(39.6,19.8 and 9.9 g/kg),with 10 rats in each group.They were given corresponding solutions for gavage for 90 consecutive days.The general condition of rats was observed,HE staining was used to observe the morphology gastric mucosa,immunofluorescence staining was used to detect the expression of PCNA protein in gastric tissue,transcriptomics obtains differentially expressed mRNA in gastric tissue and enriches differentially expressed pathways,ELISA was used to detect the contents of Bcl-xL,C-myc and Cyclin D1 in gastric tissue,RT-qPCR was used to detect the expression of Bcl-xL,C-myc and Cyclin D1 mRNA in gastric tissue,Western blot was used to detect the expressions of IL-6,JAK2,STAT3,p-JAK2 and p-STAT3 protein in gastric tissue.Results Compared with the blank group,rats in the model group showed decreased body mass(P<0.05),with structural disorders of the gastric mucosa,the expression of PCNA protein in gastric tissue increased(P<0.05),the contents and mRNA expression Bcl-xL,C-myc and Cyclin D1 in gastric tissue significantly increased(P<0.05),the expression of IL-6,JAK2,STAT3,p-JAK2,p-STAT3 protein significantly increased(P<0.05).Compared with the model group,the body mass of rats in Shenqi Yiliu Prescription high-and medium-dosage groups increased to varying degrees(P<0.05),the abnormal morphology of the gastric mucosa were improved to different degrees,and the expression of PCNA protein in Shenqi Yiliu Prescription high-,medium-and low-dosage groups significantly decreased(P<0.05).The results of transcriptomics experiments confirmed that the JAK-STAT signalling pathway showed significant differences between the blank group and model group,as well as the model group and Shenqi Yiliu Prescription high-dosage group.The content and mRNA expression of Bcl-xL,C-myc and Cyclin D1 in gastric tissue of Shenqi Yiliu Prescription high-and medium-dosage groups significantly decreased(P<0.05),and the protein expression of IL-6,JAK2,STAT3,p-JAK2 and p-STAT3 significantly decreased(P<0.05).Conclusion Shenqi Yiliu Prescription can improve the abnormal morphology of gastric mucosa in PLGC model rats,and its mechanism is related to regulating the IL-6/JAK2/STAT3 signaling pathway,thereby inhibiting cell proliferation.