β-谷甾醇和豆甾醇对小鼠急性结肠炎的治疗作用研究
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摘要
为研究β-谷甾醇和豆甾醇对由葡聚糖酸钠(DSS)诱导C57BL/6J雄性小鼠结肠炎的抑制作用。将20只小鼠随机分为4组,分别为阴性对照组、DSS处理组(1.5%DSS处理7 d),DSS+β-谷甾醇治疗组以及DSS+豆甾醇治疗组(植物甾醇溶于纯净玉米油40 mg/mL,按400 mg/kg/day的剂量灌胃小鼠)。结果显示:经过13 d的试验,DSS处理使小鼠出现严重的急性结肠炎,伴随有明显溃疡、结肠上皮组织损伤及结肠短缩等现象。豆甾醇能显著抑制结肠缩短,降低粪便中的血红素含量,降低结肠炎病理学评分(P <0.05)。并且显著降低了结肠组织炎症因子IL-1β、IL-6、MCP-1和环氧合酶COX-2的mRNA表达。β-谷甾醇也显著降低了结肠炎病理学评分,但只显著抑制IL-6和COX-2的表达。豆甾醇比β-谷甾醇表现出了更好的抗急性结肠炎活性。饮食中摄入β-谷甾醇和豆甾醇能有效改善DSS引起的小鼠结肠炎,但其在人体中的作用有待进一步调查研究。
To study the inhibiting effect of β-sitosterol and stigmasterol on dextran sulfate sodium(DSS)-induced colitis in C57 BL/6 J mice. About 20 mice were randomly divided into 4 groups, and maintained on the negative control group, DSS treatment group(1.5% DSS in water for 7 days), DSS + β-sitosterol treatment group, and DSS +stigmasterol treatment group(phytosterol in pure corn oil 40 mg/mL, given by i. g. with the dose of 400 mg/kg/day).After 13-day trial, DSS treatment induced severe acute colitis, with a marked distortion and injury of colonic epithelial and also shortening of colon. The stigmasterol significantly inhibited colon shortening, lowered the content of hemoglobin in feces, and reduced the histopathological score of colitis in the colon(P <0. 05). Stigmaterol significantly decreased the expression of several pro-inflammatory cytokines such as interleukin(IL)-1β, IL-6, monocyte chemotactic protein-1(MCP-1) and also enzyme cyclooxygenase(COX-2). β-Sitosterol also significantly reduced the severity of colitis, but it only decreased the expression of COX-2 and IL-6. Stigmasterol has higher anti-inflammatory properties than that of the β-Sitosterol. These results indicate that dietary intake of β-sitosterol and stigmasterol can effectively ameliorate colitis in mice,but their activity and in humans and the effect in human body remain to be investigated.
To study the inhibiting effect of β-sitosterol and stigmasterol on dextran sulfate sodium(DSS)-induced colitis in C57 BL/6 J mice. About 20 mice were randomly divided into 4 groups, and maintained on the negative control group, DSS treatment group(1.5% DSS in water for 7 days), DSS + β-sitosterol treatment group, and DSS +stigmasterol treatment group(phytosterol in pure corn oil 40 mg/mL, given by i. g. with the dose of 400 mg/kg/day).After 13-day trial, DSS treatment induced severe acute colitis, with a marked distortion and injury of colonic epithelial and also shortening of colon. The stigmasterol significantly inhibited colon shortening, lowered the content of hemoglobin in feces, and reduced the histopathological score of colitis in the colon(P <0. 05). Stigmaterol significantly decreased the expression of several pro-inflammatory cytokines such as interleukin(IL)-1β, IL-6, monocyte chemotactic protein-1(MCP-1) and also enzyme cyclooxygenase(COX-2). β-Sitosterol also significantly reduced the severity of colitis, but it only decreased the expression of COX-2 and IL-6. Stigmasterol has higher anti-inflammatory properties than that of the β-Sitosterol. These results indicate that dietary intake of β-sitosterol and stigmasterol can effectively ameliorate colitis in mice,but their activity and in humans and the effect in human body remain to be investigated.
引文
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[7]郭咪咪,王瑛瑶,栾霞,等.植物甾醇的提取、生理功能及在食品中的应用综述[J].食品安全质量检测学报,2014,5(9):2771-2775GU M M, WANG Y Y, LUAN X, et al. Review of the extraction, physiological function and application in food of phytosterol[J]. Journal of Food Safety Quality Inspection,2014, 5(9):2771-2775
[8] FENG S, LIU S B, LUO Z, et al. Direct saponification preparation and analysis of free and conjugated phytosterols in sugarcane(Saccharum officinarum L)by reversed-phase high-performance liquid chromatography[J]. Food Chemistry, 2015, 181:9-14
[9] KIM K A, LEE I A and GU W, et al p-Sitosterol attenuates high-fat diet-induced intestinal inflammation in mice by inhibiting the binding of lipopolysaccharide to toll-like receptor 4 in the NF-kappa B pathway[J]. Molecular Nutrition&Food Research, 2014, 58(5):963-972
[10]LEE I A, KIM E J and KIM D H. Inhibitory effect of beta-sitosterol on TNBS-induced colitis in mice[J]. Planta Medica, 2012, 78(9):896-898
[11]CHEON J H, KIM J S, KIM J M, et al. Plant sterol guggulsterone inhibits nuclear factor-kappa B signaling in intestinal epithelial cells by blocking I kappa B kinase and ameliorates acute murine colitis[J]. Inflammatory bowel diseases, 2006, 12(12):1152-1161
[12]李辉.米糠谷维素抗急性肠炎的功能评价及初步机理研究[D].长沙:中南林业科技大学,2013:37-41LI H. Progress in the Anticancer studies ofγ-tocopherol[D]. Changsha:South Central Forestry S&T university,2013:37-41
[13]ELMASHADL M N,DINA H Z,HYPERLINK EIMAN A H, et al. Immunohistochemical expression of proinflammatory enzyme COX-2 and p53 in ulcerative colitis and its associated dysplasia and colorectal carcinoma[J]. Journal of Microscopy and Ultrastructure, 2016, 4(4):195-202
[14]史红,程丽艳,郑晓亮,等.植物甾醇对慢性非细菌性前列腺炎的保护作用及其机制研究[J].中国中药杂志,2010,35(22):3033-3037SHI H, CHENG Y L, ZHENG X L, et al. Study of efects and mechanism of phytosterols on chronic abacterial prostatitis[J]. Journal of Chinese Materia Medica, 2010, 35(22):3033-3037
[15]陈超.没药甾酮抑制RAW264.7细胞炎症反应的机制研究[D].福州:福建师范大学,2011:21-33GHEN C. Inhibition of RAW264.7 macrophage inflammatory cytokines release by Guggulsterone[D]. Fuzhou:Fujian normal university, 2011, 21-33
[16] WEN X D, WANG C Z, YU C H, et al. Panax notoginseng attenuates experimental colitis in the azoxymethane/dextran sulfate sodium mouse model[J]. Phytotherapy research,2014, 28(6):892-898
[17]武晓云.甘蔗糖厂滤泥中植物甾醇的提取及豆甾醇和β-谷甾醇的分离[D].南宁:广西大学,2007:12-14WU X Y. Extraction of phytosterols from filter mud of sugarcane factory and seperation of stigmasterol andβ-sitosterol[D]. Nanning:Guangxi university, 2007:12-14
[18]田媛媛.谷维素抑制小鼠结肠炎症及其分子机理研究[D].长沙:中南林业科技大学,2014:58-59TIAN Y Y. Anti-inflammatory effects of oryzanol in colitis induced by dextran sulphate sodium(DSS)in mice and its mechanism study[D]. Changsha:Central South University of Forestry and Technology, 2014:58-59
[19]ECKMAN L, STENSON W F, SAVIDGE T C, et al. Role of intestinal epithelial cells in the host secretory response to infection by invasive bacteria-Bacterial entry induces epithelial prostaglandin h synthase-2 expression and prostaglandin E-2 and F-2 alpha production.[J]. Journal of Clinical Investigation, 1997, 100(2):296-309
[20]陆红,戈之铮,刘文忠,等.环氧合酶-2蛋白在结肠疾病中的表达[J].胃肠病学,2000,5(3):158-160LU H, GE Z Z, LIU W Z, et al. Expression of cyclooxygenase-2 protein in colon diseases[J]. Journal of Gastroenterology, 2000, 5(3):158-160
[21]李弼民,吕农华,谢勇,等.NF-κB p65反义寡核苷酸对TNBS结肠炎小鼠肠黏膜细胞因子和NF-κB表达的影响[J].世界华人消化杂志,2008,16(17):1926-1931LI B M, LU N H, XIE Y, et al. The effect of NF-κB p65 antisense oligonucleotide on intestinal mucosal cytokines and NF-κB expression in TNBS colitis mice. World Chinese Journal of Digestology, 2008, 16(17):1926-1931
[22]OH H, GRINBERG-BLEYERr Y, LIAO W, et al. An NF-κB transcription-factor-dependent lineage-specific transcriptional program promotes regulatory T cell identity and function[J]. Immunty,2017, 47(3):450-465
[23]黄松明,张爱华,丁桂霞,等.NF-KB/IKB信号通路介导血管紧张素n诱导的系膜细胞促炎性细胞因子表达[J].南京医科大学学报(自然科学版),2003,23(6):530-532HUANG S M, ZHANG A H, DING G X, et al. NF-κB/IκB Signal pathway regulated AngⅡ-indueed Proin flammatoyr Cytokines Synthesis[J]. Journal of Nanjing Medical University(Natural Science Edition), 2003, 23(6):530-532
[24]马天宇,富光明,俞腾飞.核因子-κB与溃疡性结肠炎相关性的研究进展[J].国际消化病杂志,2014,34(5):307-309+312MA T Y, FU G M, YU T F, et al. Research progress on the correlation of NF-κB with ulcerative colitis[J]. International Journal of Digestive Diseases, 2014, 34(5):307-309+312
[25]张超贤,郭李柯,郭晓凤.三硝基苯磺酸/乙醇灌肠液诱导大鼠溃疡性结肠炎TLR4/NF-κB和PI3K/AKT/NF-κB信号通路的表达及电针的干预作用[J].西安交通大学学报(医学报),2015,36(2):263-270ZHANG C X, GUO L K, GUO X F. The expressions of toll-like receptor 4/NF-κB and PI3K/AKT/NF-κB signaling pathways in ratulcerative colitis induced by the combinedema of trinitrobenzene sulphonic acid and ethano and theinterventional effect of electroacupuncture on them[J].Journal of Xi'an Jiaotong University(Medical Journal),2015, 36(2):263-270.
[2]柏亚妹,宋玉磊,卞秋桂,等.炎症性肠病患者服药依从性及影响因素分析研究[J].中国全科医学,2017,20(32):4018-4022BAI Y M, SONG Y L, BIAO Q G, et al. Medication adherence and its influencing factors among inflammatory bowel disease patients[J]. Chinese General Practice, 2017, 20(32):4018-4022
[3]赵平,董蕾,罗金燕,等.DSS与TNBS诱导大鼠实验性结肠炎模型的对比研究[J].胃肠病学,2015,20(11):667-671ZHAO P, DONG L, LUO JY, et al. Experimental colitis model induced by DSS and TNBS inrats[J]. Journal of Gastroenterology, 2015, 20(11):667-671
[4]华玉婷,李先平,王淑京,等.小鼠肠道粘膜固有层Lineage~-CD127~+淋巴细胞参与DSS诱导的早期急性结肠炎[J].中国人兽共患病学报,2016,32(11):959-964+1033HUA Y L, LI X P, WANG S J,et al. Lineage~-CD127~+lymphocytes in mouse intestinal lamina propria are involved in the earlyacute colitis induced by DSS[J]. Chinese Journal of Zoonoses, 2016, 32(11):959-964+1033
[5]饶艳霞,陈洁,吴亦栋,等.DSS诱导溃疡性结肠炎大鼠模型肝脏紧密连接蛋白的表达改变[J].临床儿科杂志,2012,30(10):967-972RAO Y X, CHEN J, WU YD, et al. Changes in the expression of liver tight junction protein in dextran sulfate sodium induced ulcerative colitis rat model[J]. The Journal of Clinical Pediatrics, 2012, 30(10):967-972
[6]张斌,郁听,栗磊,等.植物甾醇的研究进展[J].食品与发酵工业,2015,41(1):190-195ZHANG B, YU T, SU L, et al. The research progress of phytosterol[J]. Food and Fermentation Industry, 2015, 41(1):190-195
[7]郭咪咪,王瑛瑶,栾霞,等.植物甾醇的提取、生理功能及在食品中的应用综述[J].食品安全质量检测学报,2014,5(9):2771-2775GU M M, WANG Y Y, LUAN X, et al. Review of the extraction, physiological function and application in food of phytosterol[J]. Journal of Food Safety Quality Inspection,2014, 5(9):2771-2775
[8] FENG S, LIU S B, LUO Z, et al. Direct saponification preparation and analysis of free and conjugated phytosterols in sugarcane(Saccharum officinarum L)by reversed-phase high-performance liquid chromatography[J]. Food Chemistry, 2015, 181:9-14
[9] KIM K A, LEE I A and GU W, et al p-Sitosterol attenuates high-fat diet-induced intestinal inflammation in mice by inhibiting the binding of lipopolysaccharide to toll-like receptor 4 in the NF-kappa B pathway[J]. Molecular Nutrition&Food Research, 2014, 58(5):963-972
[10]LEE I A, KIM E J and KIM D H. Inhibitory effect of beta-sitosterol on TNBS-induced colitis in mice[J]. Planta Medica, 2012, 78(9):896-898
[11]CHEON J H, KIM J S, KIM J M, et al. Plant sterol guggulsterone inhibits nuclear factor-kappa B signaling in intestinal epithelial cells by blocking I kappa B kinase and ameliorates acute murine colitis[J]. Inflammatory bowel diseases, 2006, 12(12):1152-1161
[12]李辉.米糠谷维素抗急性肠炎的功能评价及初步机理研究[D].长沙:中南林业科技大学,2013:37-41LI H. Progress in the Anticancer studies ofγ-tocopherol[D]. Changsha:South Central Forestry S&T university,2013:37-41
[13]ELMASHADL M N,DINA H Z,HYPERLINK EIMAN A H, et al. Immunohistochemical expression of proinflammatory enzyme COX-2 and p53 in ulcerative colitis and its associated dysplasia and colorectal carcinoma[J]. Journal of Microscopy and Ultrastructure, 2016, 4(4):195-202
[14]史红,程丽艳,郑晓亮,等.植物甾醇对慢性非细菌性前列腺炎的保护作用及其机制研究[J].中国中药杂志,2010,35(22):3033-3037SHI H, CHENG Y L, ZHENG X L, et al. Study of efects and mechanism of phytosterols on chronic abacterial prostatitis[J]. Journal of Chinese Materia Medica, 2010, 35(22):3033-3037
[15]陈超.没药甾酮抑制RAW264.7细胞炎症反应的机制研究[D].福州:福建师范大学,2011:21-33GHEN C. Inhibition of RAW264.7 macrophage inflammatory cytokines release by Guggulsterone[D]. Fuzhou:Fujian normal university, 2011, 21-33
[16] WEN X D, WANG C Z, YU C H, et al. Panax notoginseng attenuates experimental colitis in the azoxymethane/dextran sulfate sodium mouse model[J]. Phytotherapy research,2014, 28(6):892-898
[17]武晓云.甘蔗糖厂滤泥中植物甾醇的提取及豆甾醇和β-谷甾醇的分离[D].南宁:广西大学,2007:12-14WU X Y. Extraction of phytosterols from filter mud of sugarcane factory and seperation of stigmasterol andβ-sitosterol[D]. Nanning:Guangxi university, 2007:12-14
[18]田媛媛.谷维素抑制小鼠结肠炎症及其分子机理研究[D].长沙:中南林业科技大学,2014:58-59TIAN Y Y. Anti-inflammatory effects of oryzanol in colitis induced by dextran sulphate sodium(DSS)in mice and its mechanism study[D]. Changsha:Central South University of Forestry and Technology, 2014:58-59
[19]ECKMAN L, STENSON W F, SAVIDGE T C, et al. Role of intestinal epithelial cells in the host secretory response to infection by invasive bacteria-Bacterial entry induces epithelial prostaglandin h synthase-2 expression and prostaglandin E-2 and F-2 alpha production.[J]. Journal of Clinical Investigation, 1997, 100(2):296-309
[20]陆红,戈之铮,刘文忠,等.环氧合酶-2蛋白在结肠疾病中的表达[J].胃肠病学,2000,5(3):158-160LU H, GE Z Z, LIU W Z, et al. Expression of cyclooxygenase-2 protein in colon diseases[J]. Journal of Gastroenterology, 2000, 5(3):158-160
[21]李弼民,吕农华,谢勇,等.NF-κB p65反义寡核苷酸对TNBS结肠炎小鼠肠黏膜细胞因子和NF-κB表达的影响[J].世界华人消化杂志,2008,16(17):1926-1931LI B M, LU N H, XIE Y, et al. The effect of NF-κB p65 antisense oligonucleotide on intestinal mucosal cytokines and NF-κB expression in TNBS colitis mice. World Chinese Journal of Digestology, 2008, 16(17):1926-1931
[22]OH H, GRINBERG-BLEYERr Y, LIAO W, et al. An NF-κB transcription-factor-dependent lineage-specific transcriptional program promotes regulatory T cell identity and function[J]. Immunty,2017, 47(3):450-465
[23]黄松明,张爱华,丁桂霞,等.NF-KB/IKB信号通路介导血管紧张素n诱导的系膜细胞促炎性细胞因子表达[J].南京医科大学学报(自然科学版),2003,23(6):530-532HUANG S M, ZHANG A H, DING G X, et al. NF-κB/IκB Signal pathway regulated AngⅡ-indueed Proin flammatoyr Cytokines Synthesis[J]. Journal of Nanjing Medical University(Natural Science Edition), 2003, 23(6):530-532
[24]马天宇,富光明,俞腾飞.核因子-κB与溃疡性结肠炎相关性的研究进展[J].国际消化病杂志,2014,34(5):307-309+312MA T Y, FU G M, YU T F, et al. Research progress on the correlation of NF-κB with ulcerative colitis[J]. International Journal of Digestive Diseases, 2014, 34(5):307-309+312
[25]张超贤,郭李柯,郭晓凤.三硝基苯磺酸/乙醇灌肠液诱导大鼠溃疡性结肠炎TLR4/NF-κB和PI3K/AKT/NF-κB信号通路的表达及电针的干预作用[J].西安交通大学学报(医学报),2015,36(2):263-270ZHANG C X, GUO L K, GUO X F. The expressions of toll-like receptor 4/NF-κB and PI3K/AKT/NF-κB signaling pathways in ratulcerative colitis induced by the combinedema of trinitrobenzene sulphonic acid and ethano and theinterventional effect of electroacupuncture on them[J].Journal of Xi'an Jiaotong University(Medical Journal),2015, 36(2):263-270.