苍术挥发油和苍术醇提物对溃疡性结肠炎模型小鼠的改善作用及其效果比较

doi:10.13481/j.1671-587X.20220519

摘要/Abstract

摘要：

目的探讨苍术挥发油和苍术醇提物对葡聚糖硫酸钠（DSS）诱导的小鼠溃疡性结肠炎（UC）的改善作用，为苍术的临床用药提供实验依据。方法 40只雄性BALB/c小鼠随机分为正常组、模型组、苍术挥发油（0.185 g·kg^－1）组、苍术醇提物（1.107 g·kg^－1）组和柳氮磺吡啶（0.250 g·kg^－1）组，每组8只。采用自由饮用3.5%DSS溶液建立UC小鼠模型。检测各组小鼠体质量和疾病活动指数（DAI）评分，解剖小鼠后检测各组小鼠结肠长度并计算脾脏系数，采用髓过氧化物酶（MPO）试剂盒检测各组小鼠结肠组织中MPO活性，HE染色观察各组小鼠结肠组织病理形态表现，实时荧光定量PCR（RT-qPCR）法检测各组小鼠结肠组织中肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）和白细胞介素6（IL-6） mRNA表达水平，免疫组织化学染色检测各组小鼠结肠组织中炎症因子阳性表达情况，阿利新蓝-过碘酸-雪夫（AB-PAS）染色观察各组小鼠结肠组织中杯状细胞数。结果与正常组比较，模型组小鼠第 7和8天时体质量降低（P<0.05），第4~8天时DAI评分升高（P<0.05），结肠长度缩短（P<0.05），脾脏系数增大（P<0.05），结肠组织中MPO活性升高（P<0.05），结肠组织损伤严重，结肠组织中TNF-α、IL-1β和IL-6 mRNA表达水平升高（P<0.05），炎症因子阳性表达增加，杯状细胞数减少。与模型组比较，苍术挥发油组、苍术醇提物组和柳氮磺吡啶组小鼠第7和8天时体质量升高，但差异无统计学意义（P>0.05），第6~8天时DAI评分降低（P<0.05），结肠长度增加（P<0.05），脾脏系数减小（P<0.05），结肠组织中MPO活性降低（P<0.05），结肠组织损伤程度减轻，结肠组织中TNF-α、IL-1β和IL-6 mRNA表达水平降低（P<0.05），炎症因子阳性表达减少，杯状细胞数增多。与苍术挥发油组比较，苍术醇提物组小鼠结肠组织中MPO活性降低，但差异无统计学意义（P>0.05）；TNF-α、IL-1β和IL-6 mRNA表达水平降低（P<0.05），杯状细胞数增多。结论苍术挥发油和苍术醇提物对UC模型小鼠均有改善作用，同等剂量下，苍术醇提物对小鼠UC的改善作用更好。

关键词: 苍术, 溃疡性结肠炎, 肿瘤坏死因子α, 白细胞介素1β, 白细胞介素6, 杯状细胞

Abstract:

Objective To investigate the improvement effects of oil and ethanol extract of Atractylodes rhizome on the dextran sodium sulfate （DSS）-induced ulcerative colitis （UC） in the mice， and to provide the experimental basis for the clinical application of Atractylodes rhizome. Methods Forty male BALB/c mice were randomly divided into normal group， model group， Atractylodes rhizome oil （0.185 g·kg^－1） group， Atractylodes rhizome ethanol extract （1.107 g·kg^－1） group and sulfasalazine （0.250 g·kg^－1） group， with 8 mice in each group.The UC mouse model was established by freely drinking 3.5% DSS solution. The body weights and disease activity index （DAI） scores of the mice in various groups were detected. After dissecting the mice， the colon lengthes were detected and the spleen coefficient of mice in various groups were calculated.The myeloperoxidase（MPO） activities in colon tissue of the mice in various groups were detected by MPO kit， the pathomorphology of colon tissue of the mice in various groups was observed by HE staining， the expression levels of the tumor neosis factor-α（TNF-α），interleukin-1β（IL-1β） and interleukin-6（IL-6） mRNA in colon tissue of the mice in various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） method，the positive expressions of inflammatory factors in colon tissue of the mice in various groups were detected by immunohistochemical staining，and alcian blue-periodic acid-Schiff（AB-PAS） staining was used to observe the number of goblet cells in colon tissue of the mice in various groups. Results Compared with normal group， the body weights of the mice in model group on the 7th and 8th days were decreased （P<0.05）， the DAI scores on the 4th－8th days were increased （P<0.05）， the colon lengthes were decreased （P<0.05）， the spleen coefficients were increased （P<0.05）， the MPO activities in colon tissue were increased （P<0.05）， the colon tissue was seriously damaged， the expression levels of TNF-α，IL-1β and IL-6 mRNA on the 4th－8th days were increased（P<0.05）， the positive expressions of inflammatory factors were increased， and the number of goblet cells was decreased. Compared with model group， the body weights of the mice in Atractylodes rhizome oil group， Atractylodes rhizome ethanol extract group and sulfasalazine group on the 7th and 8th days were increased，but the difference was not statistically significant（P>0.05）， the DAI scores on the 6th－8th days were decreased （P<0.05）， the colon lengthes were increased （P<0.05）， the spleen coefficients were decreased （P<0.05）， the MPO activities in colon tissue were decreased （P<0.05）， the injury degrees of colon tissue were alleviated，the expression levels of TNF-α，IL-1β and IL-6 mRNA in colon tissue were decreased（P<0.05）， the positive expressions of inflammatory factors were decreased， and the number of goblet cells was increased. Compared with Atractylodes rhizome oil group， the MPO activity in colon tissue of Atractylodes rhizome ethanol extract group was decreased， but the difference was not statistically significant（P>0.05）； the expression levels of TNF-α，IL-1β and IL-6 mRNA were decreased（P<0.05）， and the number of goblet cells was increased. Conclusion The oil and ethanol extract of Atractylodes rhizome have improvement effects in the UC model mice. At the same dose， the ethanol extract of Atractylodes rhizome can improve UC in the mice better.

Key words: Atractylodes rhizome, Ulcerative colitis, Tumor necrosis factor-α, Interleukin-1β, Interleukin-6, Goblet cell

中图分类号:

R574.62

林雄,瞿领航,许静,刘春莲,李水清,刘艳菊. 苍术挥发油和苍术醇提物对溃疡性结肠炎模型小鼠的改善作用及其效果比较[J]. 吉林大学学报(医学版), 2022, 48(5): 1247-1255.

Xiong LIN,Linghang QU,Jing XU,Chunlian LIU,Shuiqing LI,Yanju LIU. Improvement effects of Atractylodes rhizome oil and Atractylodes rhizome ethanol extract on ulcerative colitis model mice and comparison of their curative effects[J]. Journal of Jilin University(Medicine Edition), 2022, 48(5): 1247-1255.

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参考文献 21

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Weight change（η/%）	Fecal property	Occult blood or bloody stool	Score
1 <	Normal	Normal	0
1－5	Semi forming	Occult blood positive（+）	1
5－10	Mushy	Occult blood positive（?）	2
10－15	Thin	Bloody stool（+）	3
> 15	Liquid	Bloody stool（?）	4

Group	Body weight
Group	（t/d） 1	2	3	4	5	6	7	8
Normal	21.35±1.08	21.61±1.28	21.63±1.51	22.11±1.60	22.60±1.39	22.70±1.36	22.89±1.41	22.91±1.49
Model	21.75±1.19	22.08±1.09	22.32±1.05	22.60±1.01	22.47±1.29	21.57±1.36	20.38±1.47^*	18.97±1.59^*
Atractylodes rhizomeoil	21.66±0.72	21.61±0.97	21.77±0.48	21.01±1.24	20.94±1.09	21.09±1.01	20.65±0.97	20.21±1.13
Atractylodes ethanol extract	21.46±1.47	21.45±1.97	22.25±2.24	22.29±2.35	22.38±2.02	21.94±2.28	21.19±2.51	20.10±2.89
Sulfasalazine	21.56±0.29	21.92±0.25	22.20±0.12	21.98±0.22	22.12±0.31	21.74±0.39	21.31±0.73	20.22±0.68

Group	DAI
Group	（t/d） 1	2	3	4	5	6	7	8
Normal	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00	0.17±0.41	0.00±0.00	0.00±0.00	0.00±0.00
Model	0.00±0.00	0.57±0.53	1.00±0.82	2.00±0.82^*	2.71±1.38^*	5.29±2.50^*	7.71±1.80^*	11.00±1.41^*
Atractylodes rhizomeoil	0.00±0.00	0.43±0.54	0.14±0.38	1.29±1.11	1.71±1.38	2.43±2.23^△	1.29±2.98^△	5.43±2.94^△
Atractylodes ethanolextract	0.00±0.00	0.50±0.93	0.13±0.35	0.63±0.52	1.13±1.13	2.88±3.31^△	2.38±2.93^△	7.00±2.67^△
Sulfasalazine	0.00±0.00	0.00±0.00	0.00±0.00	1.00±1.56	1.50±1.92	2.50±3.11^△	2.75±4.19^△	5.50±3.87^△
^*P<0.05 compared with normal group；^△P<0.05 compared with model group.

Group	Colon length(l/cm)	Spleen coefficient	MPO activity[λ_B/(U·g^－1)]
Normal	8.97±0.70	3.43±0.20	0.63±0.22
Model	5.70±0.27^*	4.40±0.55^*	2.17±0.25^*
Atractylodes rhizome oil	6.30±0.49^△	3.60±0.30^△	1.60±0.22^△
Atractylodes rhizome ethanol extract	6.48±0.48^△	3.64±0.28^△	1.48±0.12^△
Sulfasalazine	6.83±0.75^△	3.72±0.80^△	1.39±0.12^△
^*P<0.05 compared with normal group；^△P<0.05 compared with model group.

Group	TNF-α	IL-1β	IL-6
Normal	1.00±0.31	1.00±0.51	1.00±0.16
Model	2.63±0.16^*	5.69±1.36^*	53.03±15.26^*
Atractylodes rhizome oil	1.54±0.16^△	2.89±0.96^△	2.31±0.74^△
Atractylodes rhizome ethanol extract	1.22±0.16^△#	2.72±1.05^△#	1.40±0.86^△#
Sulfasalazine	1.06±0.21^△	2.36±0.70^△	4.17±1.23^△
^P<0.05 compared with normal group；^△P<0.05 compared with model group;^#P<0.05 compared with Atractylodes rhizome* oil group.