香连丸对急性溃疡性结肠炎小鼠氧化应激介导的细胞自噬的影响研究
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摘要
目的观察香连丸对急性溃疡性结肠炎大鼠氧化应激反应及其介导的细胞自噬的影响。方法 60只C57BL/6小鼠随机分为正常组、模型组、香连丸低、中、高剂量组和美沙拉嗪组。除正常对照组外,余下各组大鼠均采用DSS的方法复制UC模型。分别观察各组大鼠结肠组织病理学形态,生化法检测氧化应激指标(MDA、SOD和GSH-px)的含量,Western blotting法检测自噬相关蛋白(Beclin-1、LC3Ⅱ、LC3Ⅰ和p62)的表达。结果香连丸能够显著改善UC小鼠结肠组织形态,降低DAI及组织学损伤评分,与模型组比较差异均具有显著统计学意义(P <0.01);此外,香连丸还能显著下调结肠组织中MDA和LC3Ⅰ的表达,增加SOD及GSH-px的活性、Beclin-1、LC3Ⅱ、p62蛋白的表达及LC3Ⅱ/LC3Ⅰ的比值,与模型组比较差异均具有统计学意义(P <0.05和P <0.01)。结论香连丸可以明显降低氧化应激反应,促进细胞自噬,减轻肠道炎性损伤,促进组织修复。
Objective: To investigate the effect of Xianglian Pill on oxidative stress response and autophagy mediated by oxidative stress response in rats with acute ulcerative colitis. Methods: 60 C57 BL/6 mice were randomly divided into the control group,the model group,Xianglian Pill low,medium and high dose groups and mesalazine group. Except the control group,UC model was replicated by DSS in the rest groups. The histopathological morphology of colon was observed;the levels of oxidative stress indexes(MDA,SOD and GSH-px) were detected by biochemical method,and the expressions of autophagy-related proteins(Beclin-1,LC3Ⅱ,LC3Ⅰ and p62) were detected by Weatern blotting. Results: Xianglian Pill significantly improved the colon histomorphology,and reduced DAI and histological damage score of UC mice,with significant differences when compared with the model group(P < 0.01). In addition,Xianglian Pill could significantly down-regulate the expressions of MDA and LC3Ⅰ in colon tissue,and increase the activity of SOD,GSH-px,the expressions of Beclin-1,LC3 II and p62 proteins,and LC3Ⅱ/LC3Ⅰ ratio;the differences were significantly when compared with the model group(P < 0.05 and P < 0.01). Conclusion: Xianglian Pill can significantly reduce oxidative stress and promote autophagy,alleviate intestinal inflammatory damage and promote tissue repair.
Objective: To investigate the effect of Xianglian Pill on oxidative stress response and autophagy mediated by oxidative stress response in rats with acute ulcerative colitis. Methods: 60 C57 BL/6 mice were randomly divided into the control group,the model group,Xianglian Pill low,medium and high dose groups and mesalazine group. Except the control group,UC model was replicated by DSS in the rest groups. The histopathological morphology of colon was observed;the levels of oxidative stress indexes(MDA,SOD and GSH-px) were detected by biochemical method,and the expressions of autophagy-related proteins(Beclin-1,LC3Ⅱ,LC3Ⅰ and p62) were detected by Weatern blotting. Results: Xianglian Pill significantly improved the colon histomorphology,and reduced DAI and histological damage score of UC mice,with significant differences when compared with the model group(P < 0.01). In addition,Xianglian Pill could significantly down-regulate the expressions of MDA and LC3Ⅰ in colon tissue,and increase the activity of SOD,GSH-px,the expressions of Beclin-1,LC3 II and p62 proteins,and LC3Ⅱ/LC3Ⅰ ratio;the differences were significantly when compared with the model group(P < 0.05 and P < 0.01). Conclusion: Xianglian Pill can significantly reduce oxidative stress and promote autophagy,alleviate intestinal inflammatory damage and promote tissue repair.
引文
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[15]王文静,孙昱皓,戴敏超,等.氧化应激对脑缺血再灌注损伤后自噬调控机制[J].中国微侵袭神经外科杂志,2013,18(6):275-279.
[16]杨其彬,何泳龙,青玉凤,等.自噬相关基因Beclin-1在间歇期痛风性关节炎患者外周血单个核细胞中的表达变化及意义[J].山东医药,2018,58(37):26-29.
[17]Tomoya L,Kei O,Hiroshi N.Role of autophagy in the pathogenesis of inflammatory bowel disease[J].World Journal of Gastroenterology,2017,23(11):1944-1953.
[18]王章流,钟继红,刘英超,等.雷公藤多苷片对溃疡性结肠炎小鼠结肠巨噬细胞自噬的影响[J].中国中西医结合消化杂志,2016,24(6):422-424.
[2]马泰,孙国平,李家斌.细胞自噬的研究方法[J].生物化学与生物物理进展,2012,39(3):204-209.
[3]方健松,马媛萍,刘畅,等.自噬介导肠黏膜屏障维持肠道稳态在炎症性肠病中的作用[J].实用医学杂志,2016,32(8):1367-1369.
[4]Yuan X,Wang B,Yang L,et al.The role of ROS-induced autophagy in hepatocellular carcinoma[J].Clin Res Hepatol Gastroenterol,2018,42(4):306-312.
[5]李昂,邢雅琪,李晓霞,等.氧化应激中ROS对FOXO3a转录因子的调控作用研究进展[J].中国药理学通报,2016,32(9):1203-1207.
[6]杨佳,范恒.复方苦参汤对溃疡性结肠炎大鼠炎性反应和氧化应激的影响[J].中华中医药杂志,2017,32(8):3457-3461.
[7]申睿,刘苗,朱向东,等.溃疡性结肠炎大鼠实验模型研究进展[J].中华中医药杂志,2018,33(9):3998-4001.
[8]董艳,陆金根.香连丸对溃疡性结肠炎大鼠细胞凋亡及Bcl-2、Bax mRNA表达的影响[J].华西医学,2016,31(6):1046-1051.
[9]Arab H,Alshorbagy M,Abdallah D,et al.Telmisartan attenuates colon inflammation,oxidative perturbations and apoptosis in a rat model of experimental inflammatory bowel disease[J].PloS one,2014,9(5):e97193.
[10]郑彩华,张娜,常玉娟,等.溃结爽灌肠组方不同组分对实验性溃疡性结肠炎大鼠的氧化应激及能量代谢水平的影响[J].河北中医药学报,2018,33(3):1-6,14.
[11]谢妲.涩肠青黛散联合美沙拉秦对溃疡性结肠炎患者炎症因子及氧化应激的影响[J].中医学报,2017,32(6):1063-1066.
[12]彭书磊.健脾补肾活血法对糖尿病肾病炎症因子的影响[J].世界中西医结合杂志,2015,10(5):686-687.
[13]Zhang Z,Yang L,Wang B,et al.Protective role of liriodendrin in mice with dextran sulphate sodium-induced ulcerative colitis[J].Int Immunopharmacol,2017,52(11):203-210.
[14]Chu XQ,Wang J,Chen GX,et al.Overexpression of microR-NA-495 improves the intestinal mucosal barrier function by targeting STAT3 via inhibition of the JAK/STAT3 signaling pathway in a mouse model of ulcerative colitis[J].Pathol Res Pract,2018,214(1):151-162.
[15]王文静,孙昱皓,戴敏超,等.氧化应激对脑缺血再灌注损伤后自噬调控机制[J].中国微侵袭神经外科杂志,2013,18(6):275-279.
[16]杨其彬,何泳龙,青玉凤,等.自噬相关基因Beclin-1在间歇期痛风性关节炎患者外周血单个核细胞中的表达变化及意义[J].山东医药,2018,58(37):26-29.
[17]Tomoya L,Kei O,Hiroshi N.Role of autophagy in the pathogenesis of inflammatory bowel disease[J].World Journal of Gastroenterology,2017,23(11):1944-1953.
[18]王章流,钟继红,刘英超,等.雷公藤多苷片对溃疡性结肠炎小鼠结肠巨噬细胞自噬的影响[J].中国中西医结合消化杂志,2016,24(6):422-424.