清胰汤对急性坏死性胰腺炎大鼠基因表达谱的影响

Effects of Qing-Yi Decoction on gene expression profile in rats with acute necrotizing pancreatitis

目的探讨清胰汤（QYD）对牛磺胆酸钠诱导的急性坏死性胰腺炎（ANP）大鼠胰腺基因表达谱的影响。方法60只sD大鼠按随机数字法分为假手术组（s0组）、ANP组和QYD组，每组20只。4％牛磺胆酸钠胰胆管逆行注射复制ANP模型，s0组注射生理盐水，QYD组3次灌胃治疗（0．75ml／100g）。观测大鼠存活率、血清淀粉酶和C．反应蛋白（CRP）变化；HE染色观察胰腺、肺组织病理改变；Illumina大鼠全基因组表达谱基因芯片分析胰腺表达谱变化；荧光定量RT—PCR验证部分基因（热休克蛋白A8和热休克蛋白b1）。结果QYD组存活率较ANP组高（80％比65％，P=0．031），而血清淀粉酶、CRP明显下降[（5789±798）比（7256±1221）U／L，P=0．001；（78．6±18．5）比（126．4±24．3）mg／L，P=0．012]，Schmidt胰腺病理评分好转。与ANP组比较，QYD组筛选出575个差异基因，其中上调92个，下调483个。基因本体论（GO）功能分析涉及到转录调节因子活性负调节、氧化还原酶类活性、酶抑制剂活性等。KEGG生物学通路主要涉及丝裂原活化蛋白激酶（MAPK）信号通路、NOD受体样信号通路、细胞周期、代谢通路、氧化还原酶类活性等。定量RT．PCR（热休克蛋白A8和热休克蛋白blmRNA）验证了基因芯片结果。结论QYD可有效治疗实验性ANP，其机制涉及到MAPK信号通路、NOD受体样信号通路、细胞周期、代谢通路、氧化还原酶类活性等。

[Abstract] Objective To explore the effects of Qing-Yi Decoction （QYD） on gene expression profile in rats with sodium taurocholate-induced acute necrotizing pancreatitis （ANP）. Methods Sixty SD rats were randomly assigned to Sham operation group（group SO, n=20）, ANP group（n=20） and QYD group （n=20）. The ANP model was established by pancreatic duct retrograde injection with 4% sodium taurocholate. SO group was treated with normal saline and QYD group received intragastric injection of QYD （0.75 ml/100 g） for thrice. The survival rates and changes in serum amylase and C-reactive protein （CRP） were determined. HE staining was employed for assessment of pathological changes in the pancreas and lung tissues. Measurement of the altered pancreatic RNA expression was conducted by using Illumina whole- genome expression biochips. Changes in particular genes, namely, Hspa8 and Hspbl, were verified via quantitative reverse transcriptase polymerase chain reaction （QRT-PCR）. Results Compared with group ANP, a higher survival rate （80% vs 65%, P=0.031 ） and Schmidt pancreas pathological scores yet reducedserum amylase[ （5789±798） U/L vs （7256±1221） U/L, P=0.001] and CRPI （78.6±18.5） mg/L vs （126.4± 24.3） mg/L, P=0.012] were noted in group QYD. Of the 575 differential genes, when compared with group ANP, group QYD yielded 92 up-regulated and 483 down-regulated genes. The gene ontology was employed to analyze the negative modulation of transcription regulator, oxidoreductase and enzyme inhibitor activity. Effects of QYD on ANP were mainly linked to KEGG metabolic pathways that involved mitogen-activated protein kinase （MAPK） and NOD receptor- like signaling pathway, cell cycle, metabolic pathways and oxidoreductase activity. Changes of Hspa8 and Hspbl mRNA in microarray were verified by QRT- PCR. Conclusion QYD is effective for the treatment of experimental ANP as a consequence of altered MAPK and NOD-like receptor signaling pathways, cell cycle, metabolic pathways and oxidoreduetase activity.