[關(guān)鍵詞]
[摘要]
目的 探討蒼術(shù)-白術(shù)藥對(duì)及各單味藥醇提物對(duì)葡聚糖硫酸鈉( DSS)溶液誘導(dǎo)潰瘍性結(jié)腸炎( UC)的小鼠腸黏膜損傷的影響。方法 將81只Balb/c雄性小鼠隨機(jī)分為對(duì)照組,模型組( 3.5% DSS),蒼術(shù)醇提物( ALEE)低、高劑量(555、1 110 mg·kg-1)組,白術(shù)醇提物( AMEE)低、高劑量( 555、1 110 mg·kg-1)組,蒼術(shù)-白術(shù)藥對(duì)醇提物( AL-AMEE)低、高劑量( 555、1 110 mg·kg-1)組,柳氮磺吡啶( SASP,陽性藥,250 mg·kg-1)組,每組9只。在7 d實(shí)驗(yàn)中,除對(duì)照組外,其余所有組的小鼠均自由飲用3.5% DSS溶液,以建立UC模型。造模第2天開始ig給藥,每天1次,連續(xù)7 d。每天對(duì)小鼠的體質(zhì)量和便血狀況進(jìn)行觀察,計(jì)算疾病活動(dòng)指數(shù)( DAI)評(píng)分;測量小鼠結(jié)腸長度、計(jì)算脾臟系數(shù);采用蘇木精-伊紅染色( HE)法觀察結(jié)腸組織的病理狀態(tài),并運(yùn)用阿利新藍(lán)/過碘酸雪夫染色( AB/PAS)評(píng)估杯狀細(xì)胞的數(shù)量;通過免疫組化法檢測結(jié)腸組織中腫瘤壞死因子( TNF)-α、白細(xì)胞介素( IL)-1β、IL-6、基質(zhì)金屬蛋白酶( MMP)-2和MMP-9蛋白的表達(dá)情況,實(shí)時(shí)熒光定量PCR( qRT-PCR)法測定TNF-α、IL-1β、IL-6、結(jié)腸組織緊密連接蛋白1( ZO-1)、閉合蛋白( Occludin)mRNA的表達(dá)。結(jié)果 與模型組相比,各給藥組體質(zhì)量、結(jié)腸長度、DAI評(píng)分、杯狀細(xì)胞數(shù)目顯著增加( P<0.05、0.01),脾臟系數(shù)顯著降低、結(jié)腸病理評(píng)分顯著降低( P<0.05、0.01);結(jié)腸組織中炎癥因子TNF-α、IL-1β、IL-6轉(zhuǎn)錄水平及其蛋白表達(dá),以及金屬基質(zhì)蛋白酶MMP-2、MMP-9蛋白表達(dá)水平顯著降低( P<0.05、0.01),且緊密連接蛋白ZO-1、Occludin的mRNA表達(dá)水平顯著升高( P<0.05、0.01),AL-AMEE組的作用最明顯。結(jié)論 蒼術(shù)-白術(shù)及其組成藥味均能改善DSS誘導(dǎo)所引起的UC炎癥,緩解結(jié)腸黏膜損傷,促進(jìn)腸道屏障修復(fù),且蒼術(shù)-白術(shù)藥對(duì)療效更為顯著。
[Key word]
[Abstract]
Objective To investigate the effects of Atractylodes lancea-A. macrocephala pair and their individual components on intestinal mucosal injury in mice with ulcerative colitis (UC) induced by dextran sulfate sodium (DSS) solution. Methods Eightyone male Balb/c mice were randomly divided into a control group, a model group (3.5% DSS), low and high dose A. lancea ethanol extract (ALEE) groups (555, 1 110 mg·kg-1), low and high dose A. macrocephala ethanol extract (AMEE) groups (555, 1 110 mg·kg-1), low and high dose A. lancea-A. macrocephala ethanol extract (AL-AMEE) groups (555, 1110 mg·kg-1), and a sulfasalazine (SASP, positive drug, 250 mg·kg-1) group, with nine mice in each group. During the seven day experiment, except for the control group, all other groups of mice freely drank 3.5% DSS solution to establish the UC model. From the second day of modeling, intragastric administration was started once a day for 7 consecutive days. The body weight and bloody stool status of the mice were observed daily, and the disease activity index (DAI) score was calculated; The colon length and spleen coefficient of the mice were measured; The pathological state of the colon tissue was observed by hematoxylin-eosin staining (HE), and the number of goblet cells was evaluated by alcian blue/periodic acid-Schiff staining (AB/PAS); The expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, matrix metalloproteinase (MMP)-2 and MMP-9 proteins in the colon tissue was detected by immunohistochemistry, and the expression of TNF-α, IL-1β, IL-6, tight junction protein 1 (ZO-1), and Occludin mRNA in the colon tissue was determined by real-time fluorescence quantitative PCR (qRT-PCR). Results Compared with the model group, the body weight, colon length, DAI score, and the number of goblet cells in each treatment group were significantly increased (P < 0.05, 0.01), the spleen coefficient was significantly decreased, and the colon pathological score was significantly decreased (P < 0.05, 0.01); The transcriptional and protein expression levels of inflammatory factors TNF-α, IL-1β, IL-6, and the protein expression levels of MMP-2 and MMP-9 in the colon tissue were significantly decreased (P < 0.05, 0.01), and the mRNA expression levels of tight junction proteins ZO-1 and Occludin were significantly increased (P < 0.05, 0.01), with the AL-AMEE group showing the most significant effect. Conclusion A. macrocephala -A. lancea and their individual components can all improve the UC inflammation induced by SS, alleviate colonic mucosal injury, and promote intestinal barrier repair, and the effect of the A. macrocephala-A. lancea pair is more significant.
[中圖分類號(hào)]
R285.5
[基金項(xiàng)目]
湖北省教育廳科學(xué)研究計(jì)劃重點(diǎn)項(xiàng)目(D20232001)