

浏览全部资源
扫码关注微信
1.成都中医药大学,成都 611137
2.成都中医药大学 附属医院,成都 610075
Received:26 May 2020,
Published Online:15 September 2020,
Published:05 December 2020
移动端阅览
杨显娟,付尹,王建等.黄连-厚朴配伍对溃疡性结肠炎模型大鼠的保护作用及对凋亡因子的影响[J].中国实验方剂学杂志,2020,26(23):83-91.
YANG Xian-juan,FU Yin,WANG Jian,et al.Protective Effect of Compatibility of Coptidis Rhizoma and Magnoliae Officinalis Cortex on Rat Model of Ulcerative Colitis and Influence of Apoptosis[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(23):83-91.
杨显娟,付尹,王建等.黄连-厚朴配伍对溃疡性结肠炎模型大鼠的保护作用及对凋亡因子的影响[J].中国实验方剂学杂志,2020,26(23):83-91. DOI: 10.13422/j.cnki.syfjx.20202238.
YANG Xian-juan,FU Yin,WANG Jian,et al.Protective Effect of Compatibility of Coptidis Rhizoma and Magnoliae Officinalis Cortex on Rat Model of Ulcerative Colitis and Influence of Apoptosis[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(23):83-91. DOI: 10.13422/j.cnki.syfjx.20202238.
目的
2
研究不同剂量的单味黄连、厚朴及配伍对溃疡性结肠炎(UC)模型大鼠的保护作用及结肠B淋巴细胞瘤-2相关X蛋白(Bax),半胱氨酸的天冬氨酸蛋白水解酶(Caspase-3)及炎性细胞因子等表达的影响。
方法
2
将120只健康成年SD大鼠随机分为空白组,模型组,柳氮磺胺吡啶组,黄连2.00,1.00,0.50 g·kg
-1
组,厚朴2.00,1.00,0.50 g·kg
-1
组,黄连-厚朴配伍4.00,2.00,1.00 g·kg
-1
组,共12组,每组10只。采用2,4,6-三硝基苯磺酸/乙醇(TNBS/乙醇)制备UC模型,造模24 h后,均按10 mL·kg
-1
灌胃,1次/d,并观察造模后各组大鼠的精神、活动状态、毛发光泽、大便性状和便血情况;连续给药6 d,末次给药24 h后取结肠组织和脾脏,计算结肠质量长度比和脾脏指数,根据结肠黏膜损伤指数(CMDI)评分标准评价结肠损伤程度,采用苏木素-伊红(HE)染色进行组织病理学观察;酶联免疫吸附测定(ELISA)检测空白组、模型组,优选的黄连2.00 g·kg
-1
组,厚朴2.00 g·kg
-1
组,黄连-厚朴配伍4.00 g·kg
-1
组血清中肿瘤坏死因子-
α
(TNF-
α
),白细胞介素-6(IL-6),白细胞介素-10(IL-10),髓过氧化物酶(MPO)的表达量;蛋白免疫印迹法(Western blot)检测大鼠结肠Bax,Caspase-3蛋白的表达。
结果
2
与空白组比较,模型组大鼠萎靡少动,结肠质量长度比、脾脏指数,CMDI,结肠组织病理损伤极显著增加,血清TNF-
α
,IL-6,MPO的表达量极显著升高,而血清IL-10表达水平则极显著降低(
P
<
0.01);与模型组比较,柳氮磺胺吡啶组,黄连2.00,1.00 g·kg
-1
,厚朴2.00 g·kg
-1
,黄连-厚朴配伍3个剂量组大鼠结肠重量长度比,CMDI均明显降低(
P
<
0.05,
P
<
0.01),黄连0.50 g·kg
-1
组,厚朴0.50,1.00 g·kg
-1
组结肠质量长度比,CMDI指数与模型组差异无统计学意义,与黄连、厚朴0.50 g·kg
-1
组比较,黄连-厚朴配伍1.00 g·kg
-1
组结肠质量长度比极显著降低(
P
<
0.01);与模型组比较,柳氮磺胺吡啶组,黄连2.00 g·kg
-1
,黄连-厚朴配伍4.00 g·kg
-1
组脾脏指数明显降低(
P
<
0.05);与模型组比较,柳氮磺胺吡啶组,黄连2.00,1.00 g·kg
-1
,厚朴2.00 g·kg
-1
,黄连-厚朴配伍3个剂量组可明显改善组织病理学损伤、组织坏死的深度和范围;与模型组比较,优选的黄连2.00 g·kg
-1
组,厚朴2.00 g·kg
-1
组,黄连-厚朴配伍4.00 g·kg
-1
组血清TNF-
α
,IL-6,MPO的表达水平显著降低,血清IL-10的水平显著升高(
P
<
0.01);与空白组比较,模型组结肠Bax,Caspase-3蛋白表达量显著升高(
P
<
0.01),与模型组比较,优选的黄连2.00 g·kg
-1
组,厚朴2.00 g·kg
-1
组,黄连-厚朴配伍4.00 g·kg
-1
组结肠Bax,Caspase-3蛋白表达量显著降低(
P
<
0.01)。
结论
2
单味黄连、厚朴及黄连-厚朴配伍可能通过下调Bax,Caspase-3蛋白的表达、抑制炎性细胞因子的释放和促进抗炎因子的释放而改善TNBS/乙醇诱导的UC模型大鼠的病理损伤,发挥保护结肠黏膜组织的作用,黄连与厚朴配伍效应有优于单味药、黄连有优于厚朴的趋势。
Objective
2
To study the protective effect of different doses of single-flavored Coptis, Magnoliae Officinalis Cortex, and their compatibility on ulcerative colitis (UC) model rats and the colonic B lymphoblastoma-2 associated X protein (Bax) and cysteine-containing aspartame-3(Caspase-3) protein, inflammatory cytokines, and other expressions.
Method
2
The 120 healthy adult SD rats were randomly divided into blank group, model group, sulfasalazine group, Coptidis Rhizoma 2.00, 1.00, 0.50 g·kg
-1
group, Magnoliae Officinalis Cortex 2.00, 1.00, 0.50 g·kg
-1
group, Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00, 2.00, 1.00 g·kg
-1
group, 12 groups with 10 rats in each group. The UC model was prepared by 2,4, 6-trinitrobenzene sulfonic acid/ethanol (TNBS/ethanol). After 24 h of modeling, the rats were gavaged at 10 mL·kg
-1
for one time/d. After modeling, the mental state, activity state, hair luster, stool characteristics, and blood in the stool of each group were observed. After continuous administration for 6 days, colon tissues and spleen were taken after the last administration for 24 h. The ratio of colonic weight to length and spleen index was calculated. The degree of colonic injury was evaluated according to the colonic mucosal injury index (CMDI) score criteria. the histopathological observation was performed using hematoxylin-eosin staining (HE). The expression levels of tumor necrosis factor (TNF)-
α
, interleukin-6 (IL-6), interleukin-10 (IL-10), and myeloperoxidase (MPO) in the serum of Coptidis Rhizoma 2.00 g·kg
-1
group, Magnoliae Officinalis Cortex 2.00 g·kg
-1
group, Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00 g·kg
-1
were detected by enzyme-linked immunosorbent assay(ELISA) in blank group and model group. Western blot was used to detect the expression of Bax and Caspase-3 proteins in the colon of rats.
Result
2
Compared with blank group, rats in model group were sluggish and less active. The colon weight-length ratio, spleen index, CMDI, and colon tissue pathological damage increased significantly, and the expression of serum TNF-
α
, IL-6, and MPO increased significantly. Serum IL-10 expression levels were extremely significantly reduced (
P
<
0.01). Compared with model group, the sulfasalazine group, the Coptidis Rhizoma 2.00, 1.00 g·kg
-1
group, the Magnoliae Officinalis Cortex 2.00 g·kg
-1
group, and the three-dose groups of Coptidis Rhizoma combine with Magnoliae Officinalis Cortex, their colon weight-length ratio and CMDI were significantly reduced (
P
<
0.05,
P
<
0.01). The colon weight length ratio and CMDI index of the Coptidis Rhizoma 0.50 g·kg
-1
group, Magnoliae Officinalis Cortex 0.50 and 1.00 g·kg
-1
group were not significantly different from the model group but compared with Coptidis Rhizoma and Magnolia 0.50 g·kg
-1
group, the ratio of colon weight to length in the group of Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 1.00 g·kg
-1
group was significantly reduced (
P
<
0.01). Compared with model group, the spleen index of the sulfasalazine group, the Coptidis Rhizoma 2.00 g·kg
-1
, and the Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00 g·kg
-1
group were significantly lower
(P
<
0.05), compared with model group, the sulfasalazine group, Coptidis Rhizoma 2.00, 1.00 g·kg
-1
and Magnoliae Officinalis Cortex 2.00 g·kg
-1
, thre dose groups of Coptidis Rhizoma combine with Magnoliae Officinalis Cortex can significantly improve the depth and scope of histopathological damage and tissue necrosis. Compared with the model group, the preferred Coptidis Rhizoma 2.00 g·kg
-1
group, Magnoliae Officinalis Cortex 2.00 g·kg
-1
group, Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00 g·kg
-1
group serum TNF-
α
, IL-6, MPO expression levels are extremely significantly reduced, the level of IL-10 increased significantly (
P
<
0.01).Compared with blank group, the expression of Bax and Caspase-3 protein in the colon of model group was significantly increased (
P
<
0.01). Compared with model group, the expression of Bax and Caspase-3 protein in preferred Coptidis Rhizoma 2.00 g·kg
-1
group and Magnoliae Officinalis Cortex 2.00 g·kg
-1
group, Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00 g·kg
-1
group were significantly reduced (
P
<
0.01).
Conclusion
2
The compatibility of single-flavored Coptidis Rhizoma, Magnoliae Officinalis Cortex, and Coptidis Rhizoma combine with Magnoliae Officinalis Cortex may improve the pathology of UC model rats induced by TNBS/ethanol by down-regulating the expression of Bax and Caspase-3 protein, inhibiting the release of inflammatory cytokines and promoting the release of anti-inflammatory factors injury, it plays a role in protecting colonic mucosa. The compatibility effect of Coptidis Rhizoma and Magnoliae Officinalis Cortex is better than that of single medicine, and Coptidis Rhizoma has a tendency to be better than Magnoliae Officinalis Cortex.
UNGARO R , MEHANDRU S , ALLEN P B , et al . Ulcerative colitis [J]. The Lancet , 2017 , 389 ( 10080 ): 1756 - 1770 .
EISENSTEIN M . Ulcerative colitis:towards remission [J]. Nature , 2018 , 563 ( 7730 ): S33 - S33 .
EISENSTEIN M . Refractory ulcerative proctitis:how to treat it? [J]. Nature , 2018 , doi: 10.1016/j.bpg.2018.05.009 http://dx.doi.org/10.1016/j.bpg.2018.05.009 .
郑新光 , 傅勇 , 王建 , 等 . 黄连配厚朴对小鼠胃肠动力及止泻作用的影响 [J]. 时珍国医国药 , 2014 , 25 ( 7 ): 1585 - 1587 .
孟斌 , 张振秋 , 黄连与厚朴配伍对盐酸小檗碱在大鼠体内药动学的影响 [J]. 中成药 , 2009 , 31 ( 6 ): 943 - 945 .
郑新光 . 黄连配厚朴减副存效及部分机制研究 [D]. 成都 : 成都中医药大学 , 2014 .
ZHU L , LU M D , HONG S , et al . Qing Chang Hua Shi granule ameliorate inflammation in experimental rats and cell model of ulcerative colitis through MEK/ERK signaling pathway [J]. Biomed Pharmacother , 2019 , 116 : 108967 .
林汉杰 , 韩凌 . 四君子汤对TNBS诱导的UC大鼠结肠紧密连接蛋白表达的影响研究 [D]. 广州 : 广州中医药大学 , 2015 .
ORDÁS I , ECKMANN L , TALAMINI M , et al . Ulcerative colitis [J]. Lancet , 2012 , 380 ( 9853 ): 1606 - 1619 .
胡宗仁 , 郑文江 , 严倩 , 等 . 溃疡性结肠炎差异表达基因及中药预测的生物信息学分析 [J]. 中国中药杂志 , 2020 , 45 ( 7 ): 1684 - 1690 .
YU X T , XU Y F , HUANG Y F , et al . Berberrubine attenuates mucosal lesions and inflammation in dextran sodium sulfate-induced colitis in mice [J]. PLoS One , 2018 , 13 ( 3 ): e0194069 .
LI C , XI Y , LI S , et al . Berberine ameliorates TNBS induced colitis by inhibiting inflammatory responses and Th1/Th17 differentiation [J]. Mol Immunol , 2015 , 67 ( 2 ): 444 - 454 .
DKHIL M A , METWALY M S , AL-QURAISHY S . Berberine improves the intestinal antioxidant status of laboratory mice,Mus musculus [J]. Saudi J Biol Sci , 2017 , 24 ( 7 ): 1567 - 1573 .
ZHANG X J , YUAN Z W , QU C , et al . Palmatine ameliorated murine colitis by suppressing tryptophan metabolism and regulating gut microbiota [J]. Pharmacol Res , 2018 , 137 : 34 - 46 .
SHEN P , ZHANG Z , HE Y , et al . Magnolol treatment attenuates dextran sulphate sodium-induced murine experimental colitis by regulating inflammation and mucosal damage [J]. Life Sci , 2018 , 196 : 69 - 76 .
祁向争 , 刘洁 . 溃疡性结肠炎动物模型研究进展 [J]. 天津中医药大学学报 , 2010 , 29 ( 4 ): 220 - 222 .
CHAMANARA M , RASHIDIAN A , MEHR S E , et al . Melatonin ameliorates TNBS-induced colitis in rats through the melatonin receptors:involvement of TLR4/MyD88/NF- κ B signalling pathway [J]. Inflammopharmacology , 2019 , 27 ( 2 ): 361 - 371 .
LOPES DE OLIVEIRA G A , ALARCÓN DE LA LASTRA C , ROSILLO M Á , et al . Preventive effect of bergenin against the development of TNBS-induced acute colitis in rats is associated with inflammatory mediators inhibition and NLRP3/ASC inflammasome signaling pathways [J]. Chem Biol Interact , 2019 , 297 : 25 - 33 .
SOUBH A A , ABDALLAH D M , EL-ABHAR H S . Geraniol ameliorates TNBS-induced colitis: Involvement of Wnt/ β -catenin,p38 MAPK,NF- κ B,and PPAR γ signaling pathways [J]. Life Sci , 2015 , 136 : 142 - 150 .
葛飞 , 孙志广 . 金荞麦对溃疡性结肠炎的抗炎作用及机制研究 [D]. 南京 : 南京中医药大学 , 2016 .
陈冠儒 , 张怡 . 芪附理中灌肠方对溃疡性结肠炎大鼠血清IL-8、IL-10及肠黏膜ICAM-1表达的影响 [D]. 成都 : 成都中医药大学 , 2018 .
乔翠霞 , 曾莉 . 溃愈散调控TLR4/MyD88/NF- κ B p65信号通路治疗溃疡性结肠炎的机制研究 [D]. 南京 : 南京中医药大学 , 2019 .
SHI L , DAI Y , JIA B , et al . The inhibitory effects of Qingchang Wenzhong granule on the interactive network of inflammation,oxidative stress,and apoptosis in rats with dextran sulfate sodium‐induced colitis [J]. J Cell Biochem , 2019 , 120 ( 6 ): 9979 - 9991 .
0
Views
14
下载量
6
CSCD
Publicity Resources
Related Articles
Related Author
Related Institution
京公网安备11010802024621