这项工作得到了吉林省科学技术厅（YDZJ202201ZYTS181）的支持。

该动物程序已获得长春中医药大学伦理委员会批准（备案号：2021214）。将无特定病原体的C57BL / 6J小鼠（8-10周，体重18-22g），雄性和雌性，饲养在22°C和65%相对湿度的独立通风笼中。小鼠在适应性喂养7天后开始实验，在此期间它们可以自由获得水和饮食。
1.药物制备
<ol><li>LJZD的制备 注：所用中药购自长春中医药大学附属医院，经鉴定为正品中药（见 表...

六君子汤：预防 UC-CRC 的有效治疗方法

结直肠癌是全球最常见的癌症之一，每年约有 1,148,000 例新发病例和超过 576,000 例死亡。结直肠癌根据病因不同可分为3种类型，包括遗传性、散发性和UC-CRC31。UC等炎症性肠病患者的CRC发病率明显高于一般人群。UC 通过炎症-癌症通路刺激 CRC 的发展，这与典型的腺瘤-腺癌通路不同 6。目前UC-CRC的病因不明，主要由长期复发性慢性炎症引起，死亡率高达60%<sup class="...

结直肠癌（CRC）是一种常见的胃肠道恶性肿瘤，是全球第三大常见恶性肿瘤，也是全球第二大常见死因，占全球癌症发病率的10%，占癌症相关死亡总数的9.4%1,2。遗传因素、慢性炎症、高脂肪饮食、糖尿病和肠道菌群异常是 CRC 3,4 的危险因素。其中，炎症性肠病，尤其是溃疡性结肠炎（UC），是CRC 5,6的明显危险因素。UC相关结直肠癌（UC-CRC）是基于结直肠慢性炎症的炎症、非典型增生和癌症的过渡过程，与CRC 7,8的典型腺瘤-腺癌发展模型不同。与一般人群相比，炎症性肠病患者患CRC的风险大约高出10-40倍9。
目前，手术仍然是 CRC 的标准治疗方法，并且根据肿瘤的位置和阶段，放射治疗、全身药物治疗或两者的组合是可能的10。虽然这些传统的治疗方式已经取得了很大的进展，但由于结直肠癌的异质性和复发率高，预后较差，治疗效果不理想11,12。因此，早期发现、早期诊断、综合治疗是提高结直肠癌患者生存率的关键，关注结直肠癌向结直肠癌的转化尤为重要。多年来，中医药因其副作用有限、疗效显著而在治疗UC-CRC或慢性胃炎方面备受关注。历代著名中医在辩证法的基础上，创作了大量的经典药方，如黄旗建中汤13、四君子汤14、思神丸15等。
流君子汤（LJZD）起源于明代编纂的易雪正传著作，是中医16中的经典处方。如表1所示，LJZD由六种传统草本植物组成，包括党参（Codonopsis pilosula，Franch.）南夫。（当神）、茯苓（Schw.）、狼（涪陵）、苍术（Atractylodes macrocephala Koidz）。（白术）， Glycyrrhiza uralensis Fisch.（Gancao）、柑橘（Citrus reticulata Blanco）和Pinellia ternata（Thunb.）蒲公英（半夏），具有补气健脾、干湿化痰的作用。在现代临床实践中，常用于治疗慢性胃炎、胃溃疡和十二指肠溃疡。现代药理学研究表明，LJZD和改良的LJZD在UC和消化道癌的辅助治疗中具有较高的应用价值17,18,19。
目前UC-CRC小鼠模型的构建方法很多，但偶氮甲烷（AOM）/葡聚糖硫酸钠（DSS）诱导的小鼠模型是应用最广泛的UC-CRC模型;临床症状、形态学和病理学观察证明，该模型与人类UC-CRC20,21非常相似。其基本原理是先用化学致癌物AOM诱导致癌作用，然后连续将小鼠暴露在DSS的炎症刺激环境中，模拟肠粘膜上皮的连续损伤和修复，从而构建UC-CRC小鼠模型22。本研究的目的是在短期内通过腹腔注射AOM和DSS的循环刺激建立UC-CRC的小鼠模型，并评价该药物和LJZD对UC-CRC的分子机制，以期为UC-CRC的治疗提供科学依据。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Azoxymethane</td>
			<td>Sigma</td>
			<td>A5486</td>
			<td></td>
		</tr>
		<tr>
			<td>5-amino salicylic acid</td>
			<td>Kuihua Pharmaceuticals Group Jiamusi Luling Pharmaceutical Co., Ltd</td>
			<td>3819413</td>
			<td></td>
		</tr>
		<tr>
			<td>C57BL/6J mice</td>
			<td>Liaoning Changsheng Biotechnology Co., Ltd</td>
			<td>NO 210726210100853716</td>
			<td></td>
		</tr>
		<tr>
			<td>Cover slip</td>
			<td>Jiangsu Shitai Experimental Equipment Co., Ltd</td>
			<td>10212432C</td>
			<td></td>
		</tr>
		<tr>
			<td>DAB color development kit</td>
			<td>Jiangsu Shitai Experimental Equipment Co., Ltd</td>
			<td>2005289</td>
			<td></td>
		</tr>
		<tr>
			<td>Dewatering machine&#160;</td>
			<td>Wuhan Junjie Electronics Co., Ltd</td>
			<td>JJ-12J</td>
			<td></td>
		</tr>
		<tr>
			<td>Dextran sulfate sodium</td>
			<td>Dalian Meilun Biotechnology Co., Ltd</td>
			<td>MB5535</td>
			<td></td>
		</tr>
		<tr>
			<td>Embedding machine</td>
			<td>Wuhan Junjie Electronics Co., Ltd</td>
			<td>JB-P5</td>
			<td></td>
		</tr>
		<tr>
			<td>Hematoxylin-eosin dye</td>
			<td>Wuhan Hundred Degree Biotechnology Co., Ltd</td>
			<td>B1000</td>
			<td></td>
		</tr>
		<tr>
			<td>IL-6</td>
			<td>Jiangsu Meimian Industrial Co., Ltd</td>
			<td>MM-0163M2</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>RWD Life Science Co., Ltd</td>
			<td>R510-22-10</td>
			<td></td>
		</tr>
		<tr>
			<td>KI67 primary antibody</td>
			<td>Google Biotechnology Inc</td>
			<td>GB121141</td>
			<td></td>
		</tr>
		<tr>
			<td>Neutral gum</td>
			<td>Wuhan Hundred Degree Biotechnology Co., Ltd</td>
			<td>10004160</td>
			<td></td>
		</tr>
		<tr>
			<td>Object slide</td>
			<td>Jiangsu Shitai Experimental Equipment Co., Ltd</td>
			<td>10212432A</td>
			<td></td>
		</tr>
		<tr>
			<td>Occludin primary antibody</td>
			<td>Affnity</td>
			<td>DF7504</td>
			<td></td>
		</tr>
		<tr>
			<td>Orthostatic optical microscope</td>
			<td>Nikon</td>
			<td>Nikon Eclipse CI</td>
			<td></td>
		</tr>
		<tr>
			<td>Pathological microtome</td>
			<td>Shanghai Leica Instrument Co., Ltd</td>
			<td>RM2016</td>
			<td></td>
		</tr>
		<tr>
			<td>ZO-1 primary antibody</td>
			<td>Abcam</td>
			<td>ab221547</td>
			<td></td>
		</tr>
	</tbody>
</table>

establishment of coloproctitis cancer model in mice and evaluation of therapeutic effect of chinese medicine

结直肠癌 （CRC） 是一种常见的消化系统恶性肿瘤，已成为全球第三大常见恶性肿瘤和恶性肿瘤相关死亡的第二大原因。溃疡性结肠直肠炎 （UC） 是一种癌前病变，UC 相关 CRC （UC-CRC） 是 CRC 最常见的亚型。因此，合理的UC-CRC模型是新药研发的基石和保障。中医药（TCM）因其良好的疗效而被广泛用于治疗UC-CRC。柳君子汤（LJZD）作为中医的经典补药方，已被广泛用于治疗UC-CRC。本研究结合偶氮甲烷和硫酸葡聚糖钠建立UC-CRC模型，并给予LJZD。数据证实，LJZD可利用小鼠体重、结直肠长度、病理炎症因子、结直肠屏障功能、癌症标志物等手段有效抑制UC-CRC的癌症转化。该协议提供了一个评估中医预防和治疗UC-CRC疗效的系统。

Colorectal cancer (CRC) is a common gastrointestinal malignancy, the third most common malignancy, and the second most common cause of death in the world, accounting for 10% of the global cancer incidence and 9.4% of the total cancer-related death1,2. Genetic factors, chronic inflammation, high-fat diet, diabetes, and abnormal intestinal flora are risk factors for CRC3,4. Among them, inflammatory bowel disease, especially ulcerative coloproctitis (UC), is a clear risk factor for CRC5,6. UC-associated CRC (UC-CRC) is a transition process of inflammation, atypical hyperplasia, and cancer based on chronic inflammation of the colorectum, which is different from the typical adenoma-adenocarcinoma development model of CRC7,8. Compared with the general population, the risk of CRC is approximately 10-40 times higher in patients with inflammatory bowel disease9.
Currently, surgery is still the standard treatment for CRC, and depending on the location and stage of the tumor, radiation therapy, systemic drug therapy, or a combination of both are possible10. Although these traditional treatment modalities have made great progress, due to the high heterogeneity and recurrence rate of CRC, the prognosis is poor, and the treatment effect is not ideal11,12. Therefore, early detection, early diagnosis, and comprehensive treatment are key to improving the survival rate of CRC patients, and it is particularly important to pay attention to the transformation of UC to CRC. Over the years, traditional Chinese medicine (TCM) has attracted much attention in the treatment of UC-CRC or chronic gastritis due to its limited side effects and significant efficacy. Based on dialectical treatment, famous Chinese medicine practitioners of various generations have created a large number of classic prescriptions, such as Huangqi Jianzhong decoction13, Sijunzi decoction14, and Sishen pill15.
Liujunzi decoction (LJZD) originated from the works of Yi Xue Zheng Zhuan compiled in the Ming Dynasty and is a classic prescription in TCM16. As shown in Table 1, LJZD consists of six traditional herbs, including Codonopsis pilosula (Franch.) Nannf. (Dangshen), Poria cocos (Schw.) Wolf (Fuling), Atractylodes macrocephala Koidz. (Baizhu), Glycyrrhiza uralensis Fisch. (Gancao), Citrus reticulata Blanco (Chenpi) and Pinellia ternata (Thunb.) Breit (Banxia), which has the effect of replenishing qi and strengthening the spleen, drying dampness, and resolving phlegm. In modern clinical practice, it is often used to treat chronic gastritis, gastric ulcers, and duodenal ulcers. Modern pharmacological research has shown that LJZD and modified LJZD have high application value in the adjuvant treatment of UC and digestive tract cancer17,18,19.
At present, there are many ways to construct UC-CRC mouse models, but the azoxymethane (AOM)/dextran sulfate sodium (DSS) induced mouse model is the most widely used UC-CRC model; the clinical symptoms, morphological, and pathological observations have proved that the model is very similar to human UC-CRC20,21. The basic principle is to first induce carcinogenesis with chemical carcinogen AOM and then continuously expose mice to the inflammatory stimulation environment of DSS to simulate the continuous damage and repair of intestinal mucosal epithelium, thereby constructing a UC-CRC mouse model22. The aim of this study is to establish a mouse model of UC-CRC by intraperitoneal injection of AOM and cyclic stimulation of DSS in the short term and to evaluate the effect of the drug and the molecular mechanism of LJZD on UC-CRC in order to provide a scientific basis for the treatment of UC-CRC.

作者聚焦： 六君子汤作为结肠直肠炎癌症的中医治疗方法

小鼠结肠直肠炎癌症模型的建立及中药治疗效果评价

This study was on the prevention and treatment of coloproctitis cancer with traditional Chinese medicine. We aim to establish a mouse model for coloproctitis cancer and evaluate traditional Chinese medicine indicators. We confirmed that the Chinese medicine Liujunzi decoction is a effective treatment for coloproctitis cancer prevention.In the future, we will work to identify more sensitive biomarkers for colon and rectal cancer and screen Liujunzi decoction for therapeutic targets.

LJZD的汤剂按表1中药物的组成比例和图1A中的中药的汤剂方法制备。根据图1B所示的时间点，小鼠在第7天腹膜内注射1 mg/mL AOM，在第3、6、9周自由饮用含有2%DSS的饮用水。UC-CRC小鼠模型在第15周成功建立。同时，从第7周到第15周，通过强饲法对小鼠进行LJZD处理。数据显示，与对照组相比，UC-CRC模型组?...

Watch this Scientific Journal Video about Author Spotlight: Liujunzi Decoction as a Traditional Chinese Treatment for Coloproctitis Cancer at JoVE.com

该方案提供了由偶氮甲烷与硫酸葡聚糖钠联合诱导的溃疡性结肠直肠炎相关结直肠癌的小鼠模型。该模型用于评价中药化合物在预防和治疗结直肠癌中的疗效。

Colorectal cancer (CRC) is a common malignancy of the digestive system and has become the third most common malignancy worldwide and the second leading ...

这项研究是关于用中医预防和治疗结肠直肠炎癌症的。我们旨在建立结肠直肠炎癌症的小鼠模型并评估中医指标。我们证实了中药流君子汤是预防结肠直肠炎癌症的有效方法。未来，我们将致力于识别结肠癌和直肠癌更敏感的生物标志物，并筛选流君子汤的治疗靶点。

小鼠结肠直肠炎癌症模型的建立及中药治疗效果评价

Abstract