桑黄菌袋提取物对葡聚糖硫酸钠诱导小鼠结肠炎的防治作用
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摘要
为了将人工栽培桑黄后的废弃物菌袋应用于动物疾病防控,研究桑黄菌袋提取物(SHM)对葡聚糖硫酸钠(DSS)诱导的小鼠结肠炎的防治效果及其作用机制。结果显示:与正常对照组相比,DSS组的小鼠体质量极显著降低(P<0.01),血浆中白细胞介素1β(IL-1β)和髓过氧化物酶(MPO)的含量极显著升高(P<0.01),结肠长度极显著变短(P<0.01),结肠组织增厚水肿,大量的炎性细胞浸润,结肠组织Toll样受体4(TLR4)、肿瘤坏死因子受体相关因子6(TRAF6)、泛素结合酶13(Ubc13)、细胞核因子κB p65(NFκB p65)、肿瘤坏死因子α(TNF-α)、IL-1β、白细胞介素6(IL-6)、核苷酸结合寡聚化结构域样受体热蛋白结构域亚家族成员3(NLRP3)和凋亡相关斑点样蛋白(ASC)的基因表达极显著升高(P<0.01);与DSS组相比,给予375 mg/(kg·d) SHM的小鼠体质量显著增加(P<0.05),血浆中IL-1β和MPO含量显著降低(P<0.05),结肠长度显著增加(P<0.05),结肠组织形态得到恢复,但给予125 mg/(kg·d) SHM的小鼠上述指标变化不显著(P>0.05);与DSS组相比,给予375 mg/(kg·d) SHM的小鼠结肠中IL-1β、IL-6和NLRP3基因的表达显著降低(P<0.05)。上述结果表明,SHM在一定剂量下对DSS诱导的小鼠结肠炎有良好的防治效果,其防治作用可能是通过降低TLR-4通路下游蛋白IL-1β和IL-6的基因表达,以及调控NLRP3的表达以降低IL-1β蛋白的活化来实现的。
This experiment was conducted to investigate the control of colitis in mice induced by dextran sodium sulfate(DSS) with Phellinus igniarius medium extract(SHM), so that wasted medium of Phellinus igniarius can be applied in controlling livestock disease. As the results indicated, in contrast to the control group, the body weight of mice in DSS group significantly decreased(P<0.01), while the contents of interleukin 1 beta(IL-1β) and myeloperoxidase(MPO) in plasma significantly increased(P<0.01). The colon length was significantly shortened(P<0.01), and the colon histology became abnormal as characterized by edema and inflammatory cell infiltration. There were also sharp rises in the expression levels of genes encoding Toll-like receptor 4(TLR4), tumor necrosis factor receptor-associated factor 6(TFR6), ubiquitin-conjugating enzyme 13(Ubc13), nuclear factor kappa B p65(NFκB p65), tumor necrosis factor alpha(TNF-α), IL-1β, interleukin 6(IL-6), nucleotide-binding o￣l￣i￣g￣o￣m￣e￣r￣i￣z￣a￣t￣i￣o￣n domain like receptor family pyrin domain containing 3(NLRP3), and apoptosis-associated speck-like protein containing a CARD(ASC) in colon in DSS group(P<0.01). In contrast to DSS group, body weight of mice treated with 375 mg/(kg·d) SHM increased significantly(P<0.05), content of IL-1β and MPO in plasma decreased signifi-cantly(P<0.05), colon length extended significantly(P<0.05), while its colon histology stayed normal. However, there was no significant change when treated with 125 mg/(kg·d) SHM(P>0.05). RT-PCR results showed that expression levels of IL-1β, IL-6 and NLRP3 in colons of the 375 mg/(kg·d) SHM treated group were much lower than those of DSS group(P<0.05). All above results suggested that SHM can alleviate DSS induced colitis at a certain dose, which might be achieved by reducing the expression levels of IL-1β, IL-6 in TLR4 pathway, and deactivating IL-1β by means of NLRP3 regulation.
This experiment was conducted to investigate the control of colitis in mice induced by dextran sodium sulfate(DSS) with Phellinus igniarius medium extract(SHM), so that wasted medium of Phellinus igniarius can be applied in controlling livestock disease. As the results indicated, in contrast to the control group, the body weight of mice in DSS group significantly decreased(P<0.01), while the contents of interleukin 1 beta(IL-1β) and myeloperoxidase(MPO) in plasma significantly increased(P<0.01). The colon length was significantly shortened(P<0.01), and the colon histology became abnormal as characterized by edema and inflammatory cell infiltration. There were also sharp rises in the expression levels of genes encoding Toll-like receptor 4(TLR4), tumor necrosis factor receptor-associated factor 6(TFR6), ubiquitin-conjugating enzyme 13(Ubc13), nuclear factor kappa B p65(NFκB p65), tumor necrosis factor alpha(TNF-α), IL-1β, interleukin 6(IL-6), nucleotide-binding o￣l￣i￣g￣o￣m￣e￣r￣i￣z￣a￣t￣i￣o￣n domain like receptor family pyrin domain containing 3(NLRP3), and apoptosis-associated speck-like protein containing a CARD(ASC) in colon in DSS group(P<0.01). In contrast to DSS group, body weight of mice treated with 375 mg/(kg·d) SHM increased significantly(P<0.05), content of IL-1β and MPO in plasma decreased signifi-cantly(P<0.05), colon length extended significantly(P<0.05), while its colon histology stayed normal. However, there was no significant change when treated with 125 mg/(kg·d) SHM(P>0.05). RT-PCR results showed that expression levels of IL-1β, IL-6 and NLRP3 in colons of the 375 mg/(kg·d) SHM treated group were much lower than those of DSS group(P<0.05). All above results suggested that SHM can alleviate DSS induced colitis at a certain dose, which might be achieved by reducing the expression levels of IL-1β, IL-6 in TLR4 pathway, and deactivating IL-1β by means of NLRP3 regulation.
引文
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[11] 李荣芳,李晓文,王吉,等.细菌脂多糖及其免疫应激研究进展[J].中兽医医药杂志,2015(4):23-26
[12] XAVIER R J,PODOLSKY D K.Unravelling the pathogenesis of inflammatory bowel disease[J].Nature,2007,448(7152):427-434
[13] 李晓东,张学彦,郑磊,等.白细胞介素在溃疡性结肠炎中的作用[J].胃肠病学和肝病学杂志,2015,24(1):4-7
[14] KORDJAZY N,HAJ-MIRZAIAN A,HAJ-MIRZAIAN A,et al.Role of toll-like receptors in inflammatory bowel disease[J].Pharmacol Res,2018,129:204-215
[15] AGUILERA M,DARBY T,MELGAR S.The complex role of inflammasomes in the pathogenesis of Inflammatory Bowel Diseases:lessons learned from experimental models[J].Cytokine Growth Factor Rev,2014,25(6):715-730
[16] LIN C J,LIEN H M,CHANG H Y,et al.Biological evaluation of Phellinus linteus-fermented broths as anti-inflammatory agents[J].J Biosci Bioeng,2014,118(1):88-93
[17] 昝立峰,包海鹰,李丹花.“桑黄”类真菌中多酚物质及其生物活性研究进展[J].天然产物研究与开发,2016,28(1):147-155
[2] HSIEH M J,CHIEN S Y,CHOU Y E,et al.Hispolon from Phellinus linteus possesses mediate caspases activation and induces human nasopharyngeal carcinomas cells apoptosis through ERK1/2,JNK1/2 and p38 MAPK pathway[J].Phytomedicine,2014,21(12):1746-1752
[3] LIN C J,LIEN H M,LIN H J,et al.Modulation of T cell response by Phellinus linteus[J].J Biosci Bioeng,2016,121(1):84-88
[4] LI Y G,JI D F,ZHONG S,et al.Polysaccharide from Phellinus linteus induces S-phase arrest in HepG2 cells by decreasing calreticulin expression and activating the P27kip1-cyclin A/D1/E-CDK2 pathway[J].J Ethnopharmacol,2013,150(1):187-195
[5] LI Y G,JI D F,ZHONG S,et al.Anti-tumor effects of proteoglycan from Phellinus linteus by immunomodulating and inhibiting Reg IV/EGFR/Akt signaling pathway in colorectal carcinoma[J].Int J Biol Macromol,2011,48(3):511-517
[6] 赵国平,戴慎,陈仁寿.中药大辞典[M].上海:上海科学技术出版社,2009:2784-2785
[7] KIM G Y,ROH S I,PARK S K,et al.Alleviation of experimental septic shock in mice by acidic polysaccharide isolated from the medicinal mushroom Phellinus linteus[J].Biol Pharm Bull,2003,26(10):1418-1423
[8] KIM S H,SONG Y S,KIM S K,et al.Anti-inflammatory and related pharmacological activities of the n-BuOH subfraction of mushroom Phellinus linteus[J].J Ethnopharmacol,2004,93(1):141-146
[9] LEE J B,SHIN Y O,BAE J S,et al.Effect of Phellinus linteus,extract supplementation on cortisol and related cytokines in young male adults[J].Food Sci Biotechnol,2010,19(3):671-675
[10] SONG M,PARK H J.Anti-inflammatory effect of Phellinus linteus grown on germinated brown rice on dextran sodium sulfate-induced acute colitis in mice and LPS-activated macrophages[J].J Ethnopharmacol,2014,154(2):311-318
[11] 李荣芳,李晓文,王吉,等.细菌脂多糖及其免疫应激研究进展[J].中兽医医药杂志,2015(4):23-26
[12] XAVIER R J,PODOLSKY D K.Unravelling the pathogenesis of inflammatory bowel disease[J].Nature,2007,448(7152):427-434
[13] 李晓东,张学彦,郑磊,等.白细胞介素在溃疡性结肠炎中的作用[J].胃肠病学和肝病学杂志,2015,24(1):4-7
[14] KORDJAZY N,HAJ-MIRZAIAN A,HAJ-MIRZAIAN A,et al.Role of toll-like receptors in inflammatory bowel disease[J].Pharmacol Res,2018,129:204-215
[15] AGUILERA M,DARBY T,MELGAR S.The complex role of inflammasomes in the pathogenesis of Inflammatory Bowel Diseases:lessons learned from experimental models[J].Cytokine Growth Factor Rev,2014,25(6):715-730
[16] LIN C J,LIEN H M,CHANG H Y,et al.Biological evaluation of Phellinus linteus-fermented broths as anti-inflammatory agents[J].J Biosci Bioeng,2014,118(1):88-93
[17] 昝立峰,包海鹰,李丹花.“桑黄”类真菌中多酚物质及其生物活性研究进展[J].天然产物研究与开发,2016,28(1):147-155