维生素D治疗大鼠结肠炎模型疗效观察及免疫调节机制的初探
发布时间：2019-05-28 16:12
【摘要】：研究背景 炎症性肠病是一组发病原因尚不明确的亚急性和慢性肠道炎症性疾病,其临床类型主要包括UC和CD。现在普遍认为CD是Thl细胞异常激活引起的疾病。维生素D近年来被发现在多种自身免疫性疾病中有重要的免疫调节作用,尤其是Thl细胞介导的自身免疫疾病。维生素D对免疫的调控可以通过多个途径达到,其中之一就是调控APC上的TLRs家族的表达及功能。通过TLRs信号通路可影响初始T细胞的分化,改变Thl细胞和Treg细胞在组织中的比例。目前有国外研究小组在动物实验上使用1,25(OH)2D3干预,发现1,25(OH)2D3有抑制炎症性肠病肠道炎症的作用。但尚无使用维生素D3及其不同剂量对于IBD疗效比较的研究。 研究目的 探索维生素D3和1,25(OH)2D3对IBD动物模型的疗效及其可能的免疫机制。 研究方法 54只SD大鼠随机分为9组,每组6只,分为：空白对照组,TNBS组,5-ASA (0.4g/kg/d)组,活性VD(1,25(OH)2D30.2ug/kg/d)组,活性VD(1,25(OH)2D30.2ug/kg/d)+5-ASA(0.4g/kg/d)组,大剂量维生素D3(7500IU/d)组,大剂量维生素D3(7500IU/d)+5-ASA(0.4g/kg/d)组,小剂量维生素D3(1800IU once)组,小剂量维生素D3(1800IU once)+5-ASA(0.4g/kg/d)组。治疗9天后于第10天处死取材。采用DAI评分,大体炎症评分,组织病理评分和结肠组织MPO测定来进行炎症程度的评估。采用免疫组化的方法评估TLR4、TLR9、Foxp3在各组大鼠结肠局部中的表达。并用RT-PCR法测定大鼠结肠组织中TLR9mRNA的含量。 实验结果 1、各TNBS处理组DAI评分、大体评分、组织病理学评分和MPO测定均较空白对照组明显升高(P0.01)。各治疗组结肠组织MPO值均显著低于TNBS组(P0.01),但仍高于空白对照组。各治疗组DAI评分、大体评分和组织病理学评分较TNBS组无明显下降(P0.05)。各治疗组之间疗效无明显差异(P0.05)。大剂量VD组和大剂量VD+5-ASA组血清钙显著高于其余各组(P0.01)。大剂量VD组血清肌酐高于其余各组(P0.05)。 2、空白对照组结肠固有层和粘膜下层炎性细胞胞膜和胞质仅有少量TLR4和TLR9表达。TNBS造模组可见大量TLR4和TLR9表达,较空白组明显增多(P0.01)。在空白对照组中仅有散在的Foxp3表达,主要不均匀分布于粘膜固有层淋巴细胞的细胞核内。TNBS组中Foxp3表达较空白组明显增多(P0.01)。单纯5-ASA治疗组TLR4、TLR9和Foxp3表达较TNBS组弱(P0.05)。各维生素D干预组TLR9和Foxp3表达较TNBS组弱(P0.05),而TLR4表达较TNBS组无明显差异(P0.05)。各维生素D干预组之间TLR4、TLR9和Foxp3表达无明显差异(P0.05)。TLR9mRNA在空白对照组中表达较少。各组之间条带灰度比值无差异(P0.05)。 实验结论 1、5-ASA、维生素D3和1,25(OH)2D能缓解大鼠结肠炎症,联合使用维生素D3和5-ASA及联合使用1,25(OH)2D和5-ASA疗效无增强。维生素D3抑制炎症的作用和剂量无关。 2、正常大鼠结肠中TLR4、TLR9和Foxp3的表达较少。 3、TNBS诱导的大鼠结肠组织中TLR4、TLR9和Foxp3表达增多。与TNBS组比较,5-ASA干预后上述三者表达均可被下调；1,25(OH)2D3或维生素D3干预后TLR9和Foxp3表达减少,TLR4表达无明显差异；不同剂量维生素D3干预后TLR4、TLR9和Foxp3的表达无明显差异。联合使用1,25(OH)2D3和5-ASA、维生素D3和5-ASA较单独使用5-ASA结肠TLR4、TLR9和Foxp3的表达无明显差异。各组TLR9在mRNA层面表达无明显差异。
[Abstract]:Study Background Inflammatory bowel disease is a group of sub-acute and chronic intestinal inflammatory diseases that are not well-defined in a group, and its clinical types mainly include UC and C D. It is widely believed that CD is a disease caused by the abnormal activation of Thl cells Vitamin D has been found to be an important immunomodulatory activity in a variety of autoimmune diseases, especially Thl cell-mediated autoimmune diseases. The regulation of vitamin D to immunity can be achieved through multiple pathways, one of which is to regulate the expression and work of the TLRs on the APC Energy. The differentiation of the initial T cells can be affected by the TLRs signaling pathway, and the ratio of the Thl cells and Treg cells to the tissue is changed The results of this study were as follows:1,25 (OH) 2D3 was used to control the intestinal inflammation of inflammatory bowel disease. A study on the efficacy of vitamin D3 and its different doses in the treatment of IBD To study. Objective To explore the effect of vitamin D3 and 1,25 (OH) 2D3 on the animal model of IBD and its possible effect Immunologic machine Methods 54 SD rats were randomly divided into 9 groups, each group was divided into two groups: the blank control group, the TNBS group, the 5-ASA (0.4 g/ kg/ day) group, the active VD (1,25 (OH) 2D30.2 ug/ kg/ day) group, the active VD (1,25 (OH) 2D30.2 ug/ kg/ day) + 5-ASA (0.4 g/ kg/ day) group, the large-dose vitamin D3 (7500 IU/ d) group, the large-dose vitamin D3 (7500 IU/ d) + 5-ASA (0.4 g/ kg/ day) group, the low-dose vitamin D3 (180 0 IU on), small dose of vitamin D3 (1800 IU on) + 5-ASA (0. 4 g/ kg/ day).9 days after treatment The samples were sacrificed on Day 10. The DAI score, the gross inflammatory score, the tissue pathology score, and the colon tissue MPO assay were used. The assessment of the degree of inflammation was carried out. TLR4, TLR9 and Foxp3 were assessed by immunohistochemistry. Expression of the local part of the colon of the rat, and the TL-PCR method was used to determine the TL in the colon tissue of the rat. R9mR The results were as follows:1. The scores of DAI, the general score, the histopathological score and the MPO in the TNBS treatment group were all blank. The number of MPO in the colon of each treatment group was significantly lower than that of the TNBS group (P0.01). 01), but still higher than the blank control group. The DAI score, general score and histopathological score of each treatment group were lower than that of the TNBS There was no significant decrease in the treatment group (P0.05). There was no significant difference between the large-dose VD group and the large-dose VD + 5-ASA group. The serum of the large-dose VD group was higher than that of the other groups (P0.01). It was higher than that of the other groups (P0.05). The cells and the cytoplasm of the lamina propria and the submucosal inflammatory cells in the blank control group were higher than that of the other groups (P0.05). There is only a small amount of TLR4 and TLR9 expression. A large number of TLR4 and TLR9 expression is visible in the TNBS fabrication module, In the blank group, only the expression of Foxp3 in the blank group was significantly increased (P0.01). The expression of Foxp3 in TNBS group The number of TLR4, TLR9 and Foxp3 in the 5-ASA treatment group was significantly higher than that in the blank group (P0.01). The expression of TLR9 and Foxp3 in each vitamin D group was weaker than that of the TNBS group (P0.05), while the expression of TLR4 was higher than that of the TNBS group (P0.05). There was no significant difference in the expression of TLR4, TLR9 and Foxp3 among the groups of vitamin D (P0.05). TLR9 MRNA was expressed in the blank control group. band-to-gray ratio The results suggest that 1,5-ASA, vitamin D3 and 1,25 (OH) 2D can relieve the inflammation of the colon of the rat, and the combination of vitamin D3 and 5-ASA and the combination of 1, 25 (OH) 2D and 5-ASA with no increase in efficacy strong. Vitamin D3 inhibits the role and dose of inflammation. Normal rat colon The expression of TLR4, TLR9 and Foxp3 was less. The expression of TLR4, TLR9 and Foxp3 in intestinal tissue was increased. Compared with TNBS group, the expression of TLR4, TLR9 and Foxp3 could be reduced after 5-ASA intervention. The expression of TLR9 and Foxp3 decreased after the intervention of 1,25 (OH) 2D3 or vitamin D3, and the expression of TLR4 was not significant. There was no significant difference in the expression of TLR4, TLR9 and Foxp3. The combined use of 1,25 (OH) 2D3 and 5-ASA, vitamin D3, and 5-ASA used alone in the 5-ASA colon The expression of TLR4, TLR9 and Foxp3 was not obvious.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R-332;R574.62
本文编号：2487190
[Abstract]:Study Background Inflammatory bowel disease is a group of sub-acute and chronic intestinal inflammatory diseases that are not well-defined in a group, and its clinical types mainly include UC and C D. It is widely believed that CD is a disease caused by the abnormal activation of Thl cells Vitamin D has been found to be an important immunomodulatory activity in a variety of autoimmune diseases, especially Thl cell-mediated autoimmune diseases. The regulation of vitamin D to immunity can be achieved through multiple pathways, one of which is to regulate the expression and work of the TLRs on the APC Energy. The differentiation of the initial T cells can be affected by the TLRs signaling pathway, and the ratio of the Thl cells and Treg cells to the tissue is changed The results of this study were as follows:1,25 (OH) 2D3 was used to control the intestinal inflammation of inflammatory bowel disease. A study on the efficacy of vitamin D3 and its different doses in the treatment of IBD To study. Objective To explore the effect of vitamin D3 and 1,25 (OH) 2D3 on the animal model of IBD and its possible effect Immunologic machine Methods 54 SD rats were randomly divided into 9 groups, each group was divided into two groups: the blank control group, the TNBS group, the 5-ASA (0.4 g/ kg/ day) group, the active VD (1,25 (OH) 2D30.2 ug/ kg/ day) group, the active VD (1,25 (OH) 2D30.2 ug/ kg/ day) + 5-ASA (0.4 g/ kg/ day) group, the large-dose vitamin D3 (7500 IU/ d) group, the large-dose vitamin D3 (7500 IU/ d) + 5-ASA (0.4 g/ kg/ day) group, the low-dose vitamin D3 (180 0 IU on), small dose of vitamin D3 (1800 IU on) + 5-ASA (0. 4 g/ kg/ day).9 days after treatment The samples were sacrificed on Day 10. The DAI score, the gross inflammatory score, the tissue pathology score, and the colon tissue MPO assay were used. The assessment of the degree of inflammation was carried out. TLR4, TLR9 and Foxp3 were assessed by immunohistochemistry. Expression of the local part of the colon of the rat, and the TL-PCR method was used to determine the TL in the colon tissue of the rat. R9mR The results were as follows:1. The scores of DAI, the general score, the histopathological score and the MPO in the TNBS treatment group were all blank. The number of MPO in the colon of each treatment group was significantly lower than that of the TNBS group (P0.01). 01), but still higher than the blank control group. The DAI score, general score and histopathological score of each treatment group were lower than that of the TNBS There was no significant decrease in the treatment group (P0.05). There was no significant difference between the large-dose VD group and the large-dose VD + 5-ASA group. The serum of the large-dose VD group was higher than that of the other groups (P0.01). It was higher than that of the other groups (P0.05). The cells and the cytoplasm of the lamina propria and the submucosal inflammatory cells in the blank control group were higher than that of the other groups (P0.05). There is only a small amount of TLR4 and TLR9 expression. A large number of TLR4 and TLR9 expression is visible in the TNBS fabrication module, In the blank group, only the expression of Foxp3 in the blank group was significantly increased (P0.01). The expression of Foxp3 in TNBS group The number of TLR4, TLR9 and Foxp3 in the 5-ASA treatment group was significantly higher than that in the blank group (P0.01). The expression of TLR9 and Foxp3 in each vitamin D group was weaker than that of the TNBS group (P0.05), while the expression of TLR4 was higher than that of the TNBS group (P0.05). There was no significant difference in the expression of TLR4, TLR9 and Foxp3 among the groups of vitamin D (P0.05). TLR9 MRNA was expressed in the blank control group. band-to-gray ratio The results suggest that 1,5-ASA, vitamin D3 and 1,25 (OH) 2D can relieve the inflammation of the colon of the rat, and the combination of vitamin D3 and 5-ASA and the combination of 1, 25 (OH) 2D and 5-ASA with no increase in efficacy strong. Vitamin D3 inhibits the role and dose of inflammation. Normal rat colon The expression of TLR4, TLR9 and Foxp3 was less. The expression of TLR4, TLR9 and Foxp3 in intestinal tissue was increased. Compared with TNBS group, the expression of TLR4, TLR9 and Foxp3 could be reduced after 5-ASA intervention. The expression of TLR9 and Foxp3 decreased after the intervention of 1,25 (OH) 2D3 or vitamin D3, and the expression of TLR4 was not significant. There was no significant difference in the expression of TLR4, TLR9 and Foxp3. The combined use of 1,25 (OH) 2D3 and 5-ASA, vitamin D3, and 5-ASA used alone in the 5-ASA colon The expression of TLR4, TLR9 and Foxp3 was not obvious.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R-332;R574.62
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相关期刊论文 前3条
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2 黄继汉,黄晓晖,陈志扬,郑青山,孙瑞元;药理试验中动物间和动物与人体间的等效剂量换算[J];中国临床药理学与治疗学;2004年09期
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