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1.广州中医药大学 科学创新中心,脾胃研究所,广州 510405
2.广州中医药大学 第一临床医学院,广州 510000
李慧璇,在读硕士,从事寒热不同宿体大肠癌选择发生方式差异研究,E-mail:505594561@qq.com
* 文彬,博士,教授,博士生导师,从事肿瘤微环境与结肠癌早期启动分子机制研究,Tel:020-36585077,E-mail:wenbin@gzucm.edu.cn
收稿日期:2019-12-31,
网络出版日期:2020-07-23,
纸质出版日期:2020-11-05
移动端阅览
李慧璇,覃景春,吕智豪等.寒热不同宿体大肠癌模型建立及差异分析[J].中国实验方剂学杂志,2020,26(21):109-117.
LI Hui-xuan,QIN Jing-chun,LYU Zhi-hao,et al.Colorectal Cancer Modeling and Difference Analysis in Cold and Heat Conditions[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(21):109-117.
李慧璇,覃景春,吕智豪等.寒热不同宿体大肠癌模型建立及差异分析[J].中国实验方剂学杂志,2020,26(21):109-117. DOI: 10.13422/j.cnki.syfjx.20201964.
LI Hui-xuan,QIN Jing-chun,LYU Zhi-hao,et al.Colorectal Cancer Modeling and Difference Analysis in Cold and Heat Conditions[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(21):109-117. DOI: 10.13422/j.cnki.syfjx.20201964.
目的
2
探求机体在寒、热不同内环境压力下大肠癌发生的相关性。
方法
2
将70只(雌雄各半)Wistar大鼠随机分为正常组10只,寒模组30只,热模组30只。寒模组予冰水(0 ℃)灌胃和冷水(10 ℃)泡浴制作寒证模型,热模组予乙醇(30%)和辣椒素(0.9 g·L
-1
)溶液制作热证模型,正常组给予生理盐水灌胃,10 mL·kg
-1
·d
-1
,持续5周。寒模组、热模组于第6周制作大肠癌模型,均予DMH溶液颈后皮下注射,25 mg·kg
-1
,每周1次,连续12次。造癌期间热模组仅予30%乙醇溶液,寒模组维持造模,10 mL·kg
-1
·d
-1
,持续38周。观察各组大鼠一般状态,测定摄食量、体质量变化情况。于第27,29,32,35,38周分批取材,将肠组织进行苏木精-伊红(HE)染色,钠、钾-三磷酸腺苷酶(Na
+
K
+
-ATP),钙、镁-三磷酸腺苷酶(Ca
2+
Mg
2+
-ATP)活性、乳酸脱氢酶(LDH)活性以及琥珀酸脱氢酶(SDH)活性检测。
结果
2
寒模组大鼠出现便血、腹水等症状明显早于热模组。与正常组比较,寒模组、热模组摄食量、体质量降低。与正常组比较,寒模组第32,35周大肠长度较短(
P
<
0.05),热模组第27,29,38周Na
+
K
+
-ATP,Ca
2+
Mg
2+
-ATP活性显著升高(
P
<
0.05,
P
<
0.01);寒模组第29,35周SDH酶活性降低(
P
<
0.05,
P
<
0.01);热模组第38周SDH酶活性显著升高(
P
<
0.01);寒模组第27周LDH酶活性显著降低(
P
<
0.01);热模组第29,32周LDH活性显著升高(
P
<
0.05,
P
<
0.01)。与热模组比较,寒模组大肠质地表现出较大的脆性,纤维化加重,促纤维增生性特征表现明显;肿瘤发生率较高,肿瘤分化程度严重;寒模组第27,29,38周Na
+
K
+
-ATP,Ca
2+
Mg
2+
-ATP酶活性显著降低(
P
<
0.01);第29,38周SDH酶活性显著降低(
P
<
0.01),第32周LDH活性降低(
P
<
0.05)。
结论
2
大肠癌处于寒冷环境时,促进肿瘤发生,处于热环境时在后期亦促进肿瘤发生。
Objective
2
To explore the correlation between the occurrence of colorectal cancer and different internal environment (cold and heat).
Method
2
The 70 Wistar rats (male and female) were randomly divided into blank group (10 cases), cold model group (30 cases) and heat model group (30 cases). The cold syndrome model was made by intragastric infusion of cold water (0 ℃) and soaking in cold water (10 ℃). The heat model was made by ethanol (30%) and capsaicin solution (0.9 g·L
-1
). The blank group was given normal saline by gavage, 10 mL·kg
-1
·d
-1
, for 5 consecutive weeks. The colorectal cancer model was made by subcutaneous injection of DMH solution in the back of neck in the cold model group and heat model group at the 6
th
week, 25 mg·kg
-1
,once a week,for 12 consecutive times. During the carcinogenesis, only 30% ethanol solution was given to the heat model group, and the modeling was maintained in cold model group, 10 mL·kg
-1
·d
-1
, for 38 weeks. The general state of the rats in each group was observed, and the changes of food intake and body weight were measured. At the 27
th
, 29
th
, 32
th
, 35
th
and 38
th
weeks, samples were collected in batches. Intestinal tissues were subjected to hematoxylin-eosin staining (HE) and detection of sodium, potassium-adenosine triphosphate (Na
+
K
+
-ATP), calcium, magnesium-adenosine triphosphate (Ca
2+
Mg
2+
-ATP) activity, lactate dehydrogenase (LDH) activity and succinate dehydrogenase (SDH) activity.
Result
2
The symptoms of hematochezia and ascites in cold model group were earlier than those in heat model group. As compared with the blank group, the food intake and body weight were decreased in cold model group and heat model group. As compared with the blank group, the length of the large intestine was shorter in cold model group at the 32
nd
and 35
th
week (
P
<
0.05), the activities of Na
+
K
+
-ATP, Ca
2+
Mg
2+
-ATP increased significantly in heat model group at the 27
th
, 29
th
and 38
th
week (
P
<
0.05,
P<
0.01), the SDH enzyme activity was decreased in the cold model group at the 29
th
and 35
th
week (
P
<
0.05,
P
<
0.01), the SDH enzyme activity was significantly increased in the heat model group at the 38
th
week (
P
<
0.01). At the 27
th
week, LDH enzyme activity was significantly reduced in cold model group (
P<
0.01). The LDH activity was increased significantly in heat model group at 29
th
and 32
nd
week (
P
<
0.05,
P
<
0.01). As compared with the heat model group, the large intestine texture of the cold model rats showed greater brittleness, aggravated fibrosis, more obvious fibroproliferative characteristics, higher tumor incidence, and more serious tumor differentiation. The Na
+
K
+
-ATP, Ca
2+
Mg
2+
-ATP enzyme activities of the cold model rats were significantly reduced at 27
th
, 29
th
, and 38
th
weeks (
P<
0.01), the SDH enzyme activity was significantly reduced in the cold model rats at 29
th
and 38
th
weeks (
P
<
0.01), the LDH activity was reduced in the cold model group at 32
nd
week (
P
<
0.05).
Conclusion
2
Cold environment for colorectal cancer promotes tumorigenesis, and the hot environment can also promote tumorigenesis in a later stage.
SIEGEL R L , MILLER K D , JEMAL A . Cancer statistics, 2019 [J]. CA Cancer J Clin , 2019 , 69 ( 1 ): 7 - 34 .
李硕 . 寒热辨证与食管癌化疗方案选择相关性的研究 [J]. 结直肠肛门外科 , 2016 , 22 ( S2 ): 88 - 90 .
POLYAK K , HAVIV I , CAMPBELL I G . Co-evolution of tumor cells and their microenvironment [J]. Trends Genet , 2009 , 25 ( 1 ): 30 - 38 .
WEBER C E , KUO P C . The tumor microenvironment [J]. Eur J Surg Oncol , 2012 , 21 ( 3 ): 172 - 177 .
ALLEN M , JONES L J . Jekyll and Hyde: the role of the microenvironment on the progression of cancer [J]. J Pathol , 2011 , 223 ( 2 ): 163 - 177 .
李德新 . 中医基础理论 [M]. 北京 : 人民卫生出版社 , 2001 : 183 .
徐玉芬 , 杨新妹 , 孙校男 , 等 . 基于“同病异治”的大肠癌不同证候血清蛋白组学探索研究 [J]. 中华全科医学 , 2015 , 13 ( 4 ): 647 - 649,663,689 .
常金荣 , 文彬 , 王汝俊 , 等 . 吴茱萸碱和小檗碱对HT29细胞凋亡及端粒酶催化亚基(hTERT)表达的影响 [J]. 中药药理与临床 , 2008 , 24 ( 2 ): 6 - 8 .
文彬 , 黄秋凌 , 熊旻利 , 等 . 实验性大肠癌模型的建立及中药的干预作用 [J]. 中药药理与临床 , 2009 , 25 ( 4 ): 79 - 82 .
文彬 , 黄秋凌 , 龚艳青 , 等 . 左金丸及其主要单体成分对大肠癌的干预作用 [J]. 世界华人消化杂志 , 2009 , 17 ( 19 ): 1936 - 1941 .
文彬 , 熊旻利 , 戴伟怡 , 等 . 左金丸及反左金丸对实验性大肠癌不同时期甲基转移酶表达的影响 [J]. 世界华人消化杂志 , 2009 , 17 ( 20 ): 2074 - 2078 .
龚艳青 , 文彬 . 左金丸对大肠癌APC表达及DNA甲基转移酶活性的影响 [J]. 南昌大学学报:医学版 , 2010 , 50 ( 12 ): 46 - 49 .
许汝福 , 尹全焕 . 各组例数不等的完全随机区段分组法 [J]. 中国卫生统计 , 1993 , 10 ( 5 ): 45 - 46 .
杨万斌 , 文彬 , 张凌杭 , 等 . 大鼠胃寒证模型造模方法探索 [J]. 中国中药杂志 , 2015 , 40 ( 20 ): 4031 - 4036 .
杨万斌 , 文彬 , 张凌杭 , 等 . 大鼠胃热证模型造模方法研究 [J]. 中国中药杂志 , 2015 , 40 ( 18 ): 3644 - 3649 .
魏华 , 张昕 , 张晨虹 , 等 . 二甲肼诱发大鼠大肠癌癌前病变模型的肠道双歧杆菌数量研究 [J]. 中华肿瘤防治杂志 , 2007 , 14 ( 20 ): 1524 - 1526 .
KOKOLUS K M , CAPITANO M L , LEE C T , et al . Baseline tumor growth and immune control in laboratory mice are significantly influenced by subthermoneutral housing temperature [J]. Proc Natl Acad Sci USA , 2013 , 110 ( 50 ): 20176 - 20181 .
陈奇 . 中药药理研究方法学 [M]. 3版 . 北京 : 人民卫生出版社 , 2011 : 33 - 36 .
陈小野 . 实用中医证候动物模型学 [M]. 北京 : 北京医科大学中国协和医科大学联合出版社 , 1993 : 78 - 79 .
方肇勤 , 潘志强 , 卢文丽 , 等 . 小鼠常见证候的辨证标准 [J]. 中国中医基础医学杂志 , 2006 , 12 ( 7 ): 508 - 510 .
孙玉文 , 陈康 , 刘家强 , 等 . 中医证候研究及寒证量化方法探索 [J]. 时珍国医国药 , 2012 , 23 ( 1 ): 202 - 203 .
朱明古 , 郭文 , 袁海锋 , 等 . 芥菜子对DMH诱导大鼠大肠癌预防作用的体内研究 [J]. 中华肿瘤防治杂志 , 2012 , 19 ( 1 ): 33 - 36 .
郑杰 . 结直肠息肉和结直肠癌 [J]. 中华病理学杂志 , 2005 , 34 ( 1 ): 9 - 10 .
O'BRIEN M J , WINAWER S J , ZAUBER A G , et al . The national polyp study. Patient and polyp characteristics associated with high-grade dysplasia in colorectal adenomas [J]. Gastroenterology , 1990 , 98 ( 2 ): 371 - 379 .
张皓婷 . 基于文献资料探讨大肠癌的中医诊治 [J]. 中国医药指南 , 2019 , 17 ( 8 ): 181 - 182 .
周杰 , 段延萍 . 对《素问·阴阳应象大论》中阴阳寒热病机的认识 [J]. 河南中医药学刊 , 2000 , 15 ( 2 ): 1 - 2 .
WINBERG R A . 癌生物学 [M]. 2版 . 北京 : 科学出版社 , 2018 : 602 - 615 .
DONGRE A , WEINBERG R A . New insights into the mechanisms of epithelial-mesenchymal transition and implications for cancer [J]. Nat Rev Mol Cell Biol , 2019 , 20 ( 2 ): 69 - 84 .
DERYNCK R , WEINBERG R A . EMT and cancer: more than meets the eye [J]. Dev Cell , 2019 , 49 ( 3 ): 313 - 316 .
SHIBUE T , REINHARDT F , WEINBERG R A . Syndecan-mediated ligation of ECM proteins triggers proliferative arrest of disseminated tumor cells [J]. Cancer Res , 2019 , 79 ( 23 ): 5944 - 5957 .
王洪琦 , 张正 , 赵燕平 , 等 . 恶性肿瘤组织中HSP70、P53表达与中医热证的关系 [J]. 中国中西医结合杂志 , 2004 , 24 ( 10 ): 897 - 900 .
王海燕 , 刘億 , 葛巍 , 等 . 四神丸对慢性复发型结肠炎大鼠结肠组织 LDH、SDH、Na + -K + -ATPase 和 Ca 2+ -Mg 2+ -ATPase 活力的影响 [J]. 中国中医基础医学杂志 , 2019 , 25 ( 7 ): 928 - 931 .
刘春蕾 , 韩冰冰 , 王世军 , 等 . 黄连氯仿、乙酸乙酯提取物对实热证模型大鼠LA、PA、LDH、SDH的影响 [J]. 山东中医药大学学报 , 2012 , 36 ( 2 ): 144 - 145 .
任永申 , 赵艳玲 , 王伽伯 , 等 . 基于动物热活性检测的肾阴虚/肾阳虚模型寒热属性差异研究 [J]. 中国实验方剂学杂志 , 2010 , 16 ( 12 ): 94 - 97,101 .
罗培灿 , 文彬 , 杨万斌 , 等 . 寒、热证候大鼠大肠癌启动时ATP酶和琥珀酸脱氢酶活性的变化 [J]. 中华中医药学刊 , 2017 , 35 ( 2 ): 310 - 312,516 .
李冀 , 柴剑波 , 李胜志 , 等 . 寒、热方剂对消炎痛型胃溃疡寒、热证模型大鼠血清 GAS,MOT 含量的影响 [J]. 中国实验方剂学杂志 , 2011 , 17 ( 20 ): 170 - 172 .
CATALANO V , TURDO A , DI FRANCO S , et al . Tumor and its microenvironment: a synergistic interplay [J]. Semin Cancer Biol , 2013 , 23 ( 6 ): 522 - 532 .
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