摘要
目的观察吴茱萸次碱对葡聚糖硫酸钠（DSS）诱导的溃疡性结肠炎（UC）小鼠治疗作用，以及观察吴茱萸次碱（Rut）对溃疡性结肠炎小鼠肠道降钙素基因相关肽（CGRP）mRNA与蛋白质表达的变化情况，探讨CGRP是否在Rut保护过程中发挥作用。方法将50只SPF级BALB／c雌性小鼠随机分为五组。除了正常组外，其余四组均用DSS溶液自由饮用7d诱导UC小鼠模型，随后随机选三组分别给予Rut30mg／kg、Rut100mg／kg、泼尼松药物治疗，观察各组小鼠的体重变化、大便性状及便血情况，进行疾病活动指数（DAI）评分及组织病理学评分，应用实时荧光定量PCR方法和免疫组化方法来检测各组小鼠肠黏膜组织中的CGRPmRNA和CGRP蛋白质表达情况。结果实验总进程的第7天，给予5％DSS溶液的四组小鼠[模型组（8．9±0．9），Rut低剂量组（8．9±0．6），Rut高剂量组（8．2±0．8），泼尼松组（8．7±1．6）]与正常组（0±0）相比，DAI评分均增高，差异有统计学意义（P〈0．01）；实验总进程的第14天，Rut低剂量组、Rut高剂量组、泼尼松组小鼠DAI评分均较第7天降低（3．2±0．6、0．9±0．6、3．1±0．7比8．9±0．6、8．2±0．8、8．7±1．6，均P〈0．05），其中Rut高剂量组DAI评分在三个治疗组中最低。与正常组相比，模型组、Rut低剂量组、Rut高剂量组、泼尼松组小鼠结肠组织学评分均升高（0．2±0．4比6．9±0．9、4．5±0．9、2．8±0．8、5．7±0．7，均P〈0．01），其中Rut高剂量组组织病理学评分在三组治疗组中最低。与正常组相比，模型组、Rut低剂量组、泼尼松组小鼠肠黏膜CGRPmRNA及蛋白质表达量均降低（0．32±0．03比0．15±0．02、0．18±0．01、0．22±0．01，均P〈0．01），但正常组与Rut高剂量组CGRPmRNA及蛋白质表达量相比差异无统计学意义（0．32±0．03比0．31±0．02，P〉0．05）。CGRPmRNA及蛋白质表达与UC小鼠DAI评分及组织病理学评分呈负相关（r=-0．797、-0．819、-0．863、-0．845，均P〈0．01）。结论Rut能改善UC小鼠模型DAI评分及组织病理学评分；Rut能上调UC小鼠结肠黏膜CGRPmRNA及蛋白质的表达；CGRPmRNA及蛋白质表达与UC小鼠DAI评分及组织病理学评分呈负相关。
Objective To determine the therapic effects of rutaecarpine in dextran sodium sulfate （DSS） induced experimental colitis and explore whether the protective role of rutaecarpine is related to the synthesis and release of CGRP. Methods Fifty female BABL/c strain mice were randomly divided into untreated model control group and DSS-exposed groups. DSS-exposed groups were given administration of 5% DSS for 7 days and respectively treated with vehicle, rutacarpine（ 30 mg/kg, 100 mg/kg） , prednisone by intragastric administration from day 8 to day 14. The disease activity index （DAI） scores, the histological scores, the mRNA and protein concentrations of CGRP in colonic tissues were measured. Results On day 7, the DAI scores of the DSS-exposed groups[ vehicle group （8.9±0. 9） ,low-dose Rut group（8.9 ±0. 6）, high-dose Rut group （ 8.2 ± 0. 8 ）, prednisone group （ 8.7 ± 1.6 ）] were much higher, compared with the untreated model control group（0 ±0） （P 〈0. 01 ）. The DAI scores on day 14 of the vehicle, rutaecarpine or prednisone treated groups were respectively markedly lower than on day 7（3.2 ±0. 6,0. 9 ±0. 6,3.1 ±0. 7 vs 8.9 ±0. 6,8.2 ±0. 8,8.7 ± 1.6, P 〈0. 05）. The DAI score of mice treated with high-dose rutaecarpine was significantly lower, compared with those treated with low-dose rutaecarpine and prednisone. Compared to the untreated model control group, the histological scores in other four groups significantly increased. Comparisons of values among the post-treatment groups had statistical significance （0. 2±0.4 vs 6. 9 ±0. 9,4. 5 ±0. 9,2. 8 ±0. 8,5.7 ±0. 7,P 〈0. 01 ）, while the high-dose rutaecarpine group presented the lowest score. The colonic mucosal CGRP mRNA and CGRP protein expressions in groups receiving vehicle,low-dose rutaecarpine and prednisone were significantly reduced than those in the untreated model control group （0. 32 ± 0. 03 vs 0. 15 ± 0. 02,0.18± 0. 01,0. 22 ± 0. 01, P 〈 0. 01 ）. The CGRP mRNA and CGRP protein expressions in the untreated model control group was similar to those in the DSS ＋ high-dose rutaecarpine group with no statistic significance between them （0. 32 ±0. 03 vs 0. 31 ± 0. 02, P 〉 0. 05 ）. Pearson correlation analysis between CGRP mRNA levels, CGRP immunohistochemisty levels and DAI, histological scores showed a statistically negative relationship respectively（ r = -0. 797, -0. 819, -0. 863, - 0. 845, all P 〈 0. 01 ）. Conclusions Rutaecarpine can ameliorate the DAI scores and histological scores of ulcerative colitis in mice. Rutaecarpine can upregulate the expressions of CGRP mRNA and CGRP protein. Correlation between CGRP mRNA, CGRP protein levels and DAI scores, histological scores respectively showed a statistically negative relationship.
作者
罗丹妮
李富军
邹益友
L Luo Danni;Li Fujun;Zou Yiyou(Department of Gastroenterology, Xiangya Hospital, Central South University, Changsha 410008, China)
出处
《中华医学杂志》
CAS
CSCD
北大核心
2018年第7期533-538,共6页
National Medical Journal of China
作者简介
通信作者：李富军，Email：13787181581@163．com
