盐酸青藤碱介导Hedgehog信号通路治疗小鼠溃疡性结肠炎分子机制的研究

徐峰; 徐琳霞; 刘伟; 王珂珂; 陈彩凤; 谢丽; 柯希权

doi:10.16766/j.cnki.issn.1674-4152.002671

盐酸青藤碱介导Hedgehog信号通路治疗小鼠溃疡性结肠炎分子机制的研究

doi: 10.16766/j.cnki.issn.1674-4152.002671

蚌埠医学院第一附属医院消化内科，安徽蚌埠 233004

基金项目:

安徽省教育厅2018年度高校优秀青年骨干人才国内外访学研修项目 gxgwfx2018052

详细信息

通讯作者:
柯希权, E-mail: kexq2006@sina.com

中图分类号: R574.62 R-332
计量
- 文章访问数: 271
- HTML全文浏览量: 170
- PDF下载量: 8
- 被引次数: 0
出版历程
- 收稿日期: 2022-03-30
- 网络出版日期: 2022-11-30

The molecular mechanism of sinomenine hydrochloride treat ulcerative colitis through Hedgehog signalling pathway in mice

Department of Gastroenterology, the First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui 233004, China

摘要

摘要: 目的研究盐酸青藤碱(SH)对小鼠溃疡性结肠炎(UC)的保护作用及可能机制。方法将24只小鼠采用随机数字表法分为正常组、模型组、SH低、高剂量组(SH 20 mg/kg、60 mg/kg)4组，每组6只；记录疾病活动指数(DAI)和结肠组织病理学评分，检测Shh mRNA及相关蛋白和细胞因子的表达。结果与正常组相比，模型组DAI评分[(3.44±0.27)分vs. 0分, P＜0.001]升高，组织病理学评分升高[(7.67±0.52)分vs. 0分，P＜0.001]，Shh mRNA降低(P＜0.05)，Shh、Smo、Ptch1、Gli1蛋白表达水平降低(均P＜0.01)，TNF-α、IL-1β、IL-6升高(均P＜0.001)；与模型组相比, SH低、高剂量组DAI评分[(2.50±0.18)分、(1.89±0.17)分vs. (3.44±0.27)分, P < 0.001]降低, 组织病理学评分降低[(5.17±0.75)分、(3.33±0.52)分vs. (7.67±0.52)分, P < 0.001]，Shh、Smo、Ptch1、Gli1蛋白升高(均P＜0.001)，TNF-α、IL-1β、IL-6降低(均P＜0.01)。结论 SH可治疗小鼠UC，其机制可能是SH上调了Hedgehog信号通路活性，从而降低了TNF-α、IL-1β、IL-6含量。
- 盐酸青藤碱 /
- 溃疡性结肠炎 /
- 葡聚糖硫酸钠 /
- Hedgehog信号通路
Abstract: Objective To investigate the protective effect and possible mechanism of sinomenine hydrochloride (SH) on ulcerative colitis (UC) in mice. Methods Twenty-four mice were randomly divided into 4 groups: normal group, model group, low and high dose SH group (SH 20 mg/kg, 60 mg/kg), with 6 mice in each group. Disease activity index (DAI) and colon histopathology (HI) scores were recorded, and the expression levels of Shh mRNA and related proteins and cytokines were detected. Results Compared with the normal group, the DAI score [(3.44±0.27) points vs. 0, P < 0.001] and HI score [(7.67±0.52) points vs. 0, P < 0.001] increased, Shh mRNA (P < 0.05), Shh, Smo, Ptch1 and Gli1 protein (all P < 0.01) decreased, and TNF-α, IL-1β and IL-6 (all P < 0.001) increased in the model group. Compared with the model group, the DAI score [(2.50±0.18) points, (1.89±0.17) points vs. (3.44±0.27) points, P < 0.001], HI score [(5.17±0.75) points, (3.33±0.52) points vs. (7.67±0.52) points, P < 0.001] decreased, Shh, Smo, Ptch1 and Gli1 protein (all P < 0.001) increased, and TNF-α, IL-1β, IL-6 (all P < 0.01) decreased in the SH treatment group. Conclusion SH can treat ulcerative colitis in mice. The mechanism may be that it up regulates the activity of Hedgehog signalling pathway, thereby reduces the content of TNF-α, IL-1β, IL-6.
- Sinomenine hydrochloride /
- Ulcerative colitis /
- Dextran sodium sulfate /
- Hedgehog signalling pathway

HTML全文

图 1 各组小鼠体重下降百分比及DAI评分比较

注：A为各组小鼠体重相当于初始值百分比；B为各组小鼠DAI评分。与同时间点正常组比较，^aP＜0.001；与同时间点模型组比较，^bP＜0.001；与SH低剂量组比较，^cP＜0.001。

Figure 1. Comparison of weight loss percentage and DAI score of mice in each group

下载: 全尺寸图片幻灯片

图 2 各组小鼠HE染色情况(×100)

注：A为正常组，B为模型组，C为SH低剂量组，D为SH高剂量组。

Figure 2. HE staining of mice in each group(×100)

下载: 全尺寸图片幻灯片

图 3 各组小鼠结肠中Hedgehog通路相关蛋白表达情况

Figure 3. Expression of Hedgehog signaling pathway related proteins in colon of mice in each group

下载: 全尺寸图片幻灯片

表 1 各组小鼠结肠组织内Hedgehog信号通路Shh、Smo、Ptch1、Gli1蛋白表达水平比较(x±s)

Table 1. Comparison of the protein expression levels of Shh, Smo, Ptch1 and Gli1 of Hedgehog signaling pathway in colon tissue of mice in each group(x±s)

组别	只数	Shh	Smo	Ptch1	Gli1
正常组	6	1.00±0.00	1.00±0.00	1.00±0.00	1.00±0.00
模型组	6	0.77±0.08^a	0.80±0.09^a	0.73±0.09^a	0.74±0.10^a
SH低剂量组	6	1.44±0.10^ab	1.35±0.15^ab	1.42±0.13^ab	1.56±0.16^ab
SH高剂量组	6	1.83±0.20^abc	1.62±0.22^abc	1.81±0.09^abc	2.05±0.20^abc
F值		94.771	51.484	132.684	97.811
P值		＜0.001	＜0.001	＜0.001	＜0.001
注：与正常组比较，^aP < 0.05；与模型组比较，^bP < 0.05；与SH低剂量组比较，^cP < 0.05。

下载: 导出CSV

表 2 各组小鼠血清中细胞因子TNF-α、IL-1β、IL-6表达水平比较(x±s，pg/mL)

Table 2. Comparison of cytokine TNF-α, IL-1β, IL-6 expression levels in serum of mice in each group (x±s, pg/mL)

1组别	只数	TNF-α	IL-1β	IL-6
正常组	6	164.61±29.09	21.95±1.56	32.78±6.84
模型组	6	718.53±81.81^a	51.62±2.80^a	444.07±67.77^a
SH低剂量组	6	544.72±90.03^ab	34.10±2.90^ab	345.43±43.40^ab
SH高剂量组	6	434.11±71.75^abc	29.44±3.70^abc	236.11±29.35^abc
F值		62.258	72.606	101.150
P值		＜0.001	＜0.001	＜0.001
注：与正常组比较，^aP＜0.001；与模型组比较，^bP＜0.01；与SH低剂量组比较，^cP＜0.05。

下载: 导出CSV

参考文献(20)

[1]	FLYNN S, EISENSTEIN S. Inflammatory bowel disease presentation and diagnosis[J]. Surg Clin North Am, 2019, 99(6): 1051-1062. doi: 10.1016/j.suc.2019.08.001
[2]	徐峰, 柯希权. 溃疡性结肠炎与肠道微生物关系研究进展[J]. 胃肠病学和肝病学杂志, 2021, 30(9): 969-971. doi: 10.3969/j.issn.1006-5709.2021.09.003 XU F, KE X Q. Progress of ulcerative colitis and intestinal microorganisms[J]. Chinese Journal of Gastroenterology and Hepatology, 2021, 30(9): 969-971. doi: 10.3969/j.issn.1006-5709.2021.09.003
[3]	NG S C, SHI H Y, HAMIDI N, et al. Worldwide incidence and prevalence of inflammatory bowel disease in the 21st century: A systematic review of population-based studies[J]. Lancet, 2017, 390(10114): 2769-2778. doi: 10.1016/S0140-6736(17)32448-0
[4]	吴开春, 梁洁, 冉志华, 等. 炎症性肠病诊断与治疗的共识意见(2018年·北京)[J]. 中国实用内科杂志, 2018, 38(9): 796-813. doi: 10.19538/j.nk2018090106 WU K C, LIANG J, RAN Z H, et al. Chinese consensus on diagnosis and treatment of inflammatory bowel disease (Beijing, 2018)[J]. Chinese Journal of Practical Internal Medicine, 2018, 38(9): 796-813. doi: 10.19538/j.nk2018090106
[5]	FU Y F, LI L, FANG P, et al. Sinomenine ' s protective role and mechanism in stress load-induced heart failure[J]. J Pharm Pharmacol, 2020, 72(2): 209-217. doi: 10.1111/jphp.13181
[6]	SINGH D, AGRAWAL A, SINGAL C M S, et al. Sinomenine inhibits amyloid beta-induced astrocyte activation and protects neurons against indirect toxicity[J]. Mol Brain, 2020, 13(1): 30. doi: 10.1186/s13041-020-00569-6
[7]	GAO L N, ZHONG B, WANG Y. Mechanism underlying antitumor effects of sinomenine[J]. Chin J Integr Med, 2019, 25(11): 873-878. doi: 10.1007/s11655-019-3151-2
[8]	ZHOU Y, LIU H Y, SONG J, et al. Sinomenine alleviates dextran sulfate sodium-induced colitis via the Nrf2/NQO-1 signaling pathway[J]. Mol Med Rep, 2018, 18(4): 3691-3698.
[9]	ABRAHAM B P, QUIGLEY E M M. Probiotics in inflammatory bowel disease[J]. Gastroenterol Clin North Am, 2017, 46(4): 769-782. doi: 10.1016/j.gtc.2017.08.003
[10]	VEENSTRA J P, VEMU B, TOCMO R, et al. Pharmacokinetic analysis of carnosic acid and carnosol in standardized rosemary extract and the effect on the disease activity index of DSS-induced colitis[J]. Nutrients, 2021, 13(3): 773-789. doi: 10.3390/nu13030773
[11]	KOJIMA F, SEKIYA H, HIOKI Y, et al. Facilitation of colonic T cell immune responses is associated with an exacerbation of dextran sodium sulfate-induced colitis in mice lacking microsomal prostaglandin E synthase-1[J]. Inflamm Regen, 2022, 42(1): 1. doi: 10.1186/s41232-021-00188-1
[12]	KUCHARZIK T. Living guideline on ulcerative colitis[J]. Chirurg, 2022, 93(3): 261-265. doi: 10.1007/s00104-022-01594-y
[13]	WALTON K D, GUMUCIO D L. Hedgehog signaling in intestinal development and homeostasis[J]. Annu Rev Physiol, 2021, 83: 359-380. doi: 10.1146/annurev-physiol-031620-094324
[14]	BUONGUSTO F, BERNARDAZZI C, YOSHIMOTO A N, et al. Disruption of the Hedgehog signaling pathway in inflammatory bowel disease fosters chronic intestinal inflammation[J]. Clin Exp Med, 2017, 17(3): 351-369. doi: 10.1007/s10238-016-0434-1
[15]	EICHELE D D, KHARBANDA K K. Dextran sodium sulfate colitis murine model: An indispensable tool for advancing our understanding of inflammatory bowel diseases pathogenesis[J]. World J Gastroenterol, 2017, 23(33): 6016-6029. doi: 10.3748/wjg.v23.i33.6016
[16]	NEURATH M F, CHIRIAC M T. Targeting immune cell wiring in ulcerative colitis[J]. Immunity, 2019, 51(5): 791-793. doi: 10.1016/j.immuni.2019.10.011
[17]	江海燕, 王新月, 王建云, 等. 解毒活血灌肠方联合中药口服治疗溃疡性结肠炎患者的临床研究[J]. 中华全科医学, 2021, 19(11): 1925-1928, 1955. doi: 10.16766/j.cnki.issn.1674-4152.002202 JIANG H Y, WANG X Y, WANG J Y, et al. Clinical study of Jiedu Huoxue Decoction enema combined with oral Chinese medicine in patients with ulcerative colitis[J]. Chinese Journal of General Practice, 2021, 19(11): 1925-1928, 1955. doi: 10.16766/j.cnki.issn.1674-4152.002202
[18]	LIU W W, ZHANG Y J, ZHU W N, et al. Sinomenine inhibits the progression of rheumatoid arthritis by regulating the secretion of inflammatory cytokines and monocyte/macrophage subsets[J]. Front Immunol, 2018, 9: 2228. doi: 10.3389/fimmu.2018.02228
[19]	GAO T L, SHI T S, WIESENFELD-HALLIN Z, et al. Sinomenine facilitates the efficacy of gabapentin or ligustrazine hydrochloride in animal models of neuropathic pain[J]. Eur J Pharmacol, 2019, 854: 101-108. doi: 10.1016/j.ejphar.2019.03.061
[20]	HELAL M G, ABD ELHAMEED A G. Graviola mitigates acetic acid-induced ulcerative colitis in rats: Insight on apoptosis and Wnt/Hh signaling crosstalk[J]. Environ Sci Pollut Res Int, 2021, 28(23): 29615-29628. doi: 10.1007/s11356-021-12716-0