中国临床药理学杂志

解毒地黄汤拮抗结肠癌线粒体自噬及NLRP3活化的作用及机制研究

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李育民 ¹ , 黄灿灿 ² , 归明彬 ³ , 岳斌 ⁴ , 李兆桓 ⁴ , 王芸 ¹ , 薛秀芳 ¹ , 裴正学 ⁵

中国临床药理学杂志 | 临床与基础桥接研究 2026,42(3): 356-363

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中国临床药理学杂志 |临床与基础桥接研究 2026 , 42 (3) : 356 -363

解毒地黄汤拮抗结肠癌线粒体自噬及NLRP3活化的作用及机制研究

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李育民¹, 黄灿灿², 归明彬³, 岳斌⁴, 李兆桓⁴, 王芸¹, 薛秀芳¹, 裴正学⁵

作者信息

^1.甘肃省第二人民医院/西北民族大学附属医院肿瘤内科甘肃兰州 730030

^2.南通市中医院妇科，江苏南通 226000

^3.中国人民解放军联勤保障部队第九四〇医院结直肠肛门外科甘肃兰州 730050

^4.甘肃中医药大学中医临床学院，甘肃兰州 730000

^5.甘肃省肿瘤医院中西医结合科，甘肃兰州 730050

通讯作者:

薛秀芳，副研究员 MP: 13919770279 E-mail: 243825948@qq.com

作者简介:

李育民（1991-），男，主治医生，主要从事中西医结合防治肿瘤相关研究工作

Research on the effect and mechanism of Jiedu Dihuang Decoction in antagonizing mitochondrial autophagy and NLRP3 activation in colorectal cancer

Yu-min LI¹, Can-can HUANG², Ming-bin GUI³, Bin YUE⁴, Zhao-huan LI⁴, Yun WANG¹, Xiu-fang XUE¹, Zheng-xue PEI⁵

Affiliations

^1.Departinent of Medical Oncology, Gansu Provincial Second People’s Hospital / Affiliated Hospital of Northwest Minzu University, Lanzhou 730030, Gansu Province, China

^2.Department of Gynecology, Nantong Hospital of Traditional Chinese Medicine, Nantong 226000, Jiangsu Province, China

^3.Department of Colorectal & Anal surgery, The 940th Hospital of the Joint Logistics Support Force of Chinese People’s Liberation Army, Lanzhou 730050, Gansu Province, China

^4.Clinical College of Chinese Medicine, Gansu University of Chinese Medicine, Lanzhou 730000, Gansu Province, China

^5.Department of Integrated Traditional Chinese and Western Medicine, Gansu Provincial Cancer Hospital, Lanzhou 730050, Gansu Province, China

出版时间: 2026-02-17 doi: 10.13699/j.cnki.1001-6821.2026.03.010

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摘要

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目的

探索解毒地黄汤调控线粒体自噬与苷酸结合寡聚化域（NOD）样受体热蛋白结构域相关蛋白3（NLRP3）炎症小体对结肠癌的炎症微环境的干预效应。

方法

用腹腔注射氧化偶氮甲烷（AOM）联合饲喂葡聚糖硫酸钠（DSS）成功复制动物模型后随机分为对照组，模型组，解毒地黄汤低、中、高剂量组，阳性药物组，90只C57BL/6雄性小鼠分为6组，每组15只，各组连续处理10 w。用苏木精-伊红（HE）染色法评价直肠组织病理形态改变；用免疫荧光染色法观察结肠组织PTEN诱导激酶1（PINK1）、帕金森病蛋白（Parkin）、B细胞淋巴瘤-2蛋白相互作用中心卷曲螺旋蛋白1（Beclin1）的表达部位及强度；用聚合酶链式反应（PCR）法检测直肠NLRP3、凋亡相关斑点样蛋白（ASC）、半胱氨酸天冬氨酸蛋白酶-1（Caspase-1）mRNA的表达；用蛋白质印迹法（WB）检测PINK1、Parkin、Beclin1、NLRP3、ASC、Caspase-1蛋白表达水平；用流式细胞术检测辅助性T细胞1（Th1）与辅助性T细胞2（Th2）的比例；用酶联免疫吸附测定法（Elisa）检测白细胞介素-4（IL-4）、白细胞介素-6（IL-6）、白细胞介素-10（IL-10）、白细胞介素-12（IL-12）、肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFN-γ）的水平。

结果

模型组和对照组的NLRP3 mRNA相对表达水平分别为2.06±0.22和1.01±0.17，ASC的相对表达水平分别为3.03±0.16和1.01±0.19，Caspase-1 mRNA相对表达水平分别为3.28±0.22和1.00±0.11，在统计学上差异均具有统计学意义（均P<0.01）；模型组和对照组PINK1平均荧光强度分别为76.39±4.89和122.99±6.93，Parkin平均荧光强度分别为83.12±5.01和138.38±3.84，Beclin1平均荧光强度分别为87.43±3.77和143.22±9.42，在统计学上差异均具有统计学意义（均P<0.01）；模型组与对照组腹腔液Th1占比分别为（0.74±0.09）%和（4.78±0.62）%、Th2占比分别为（2.05±0.26）%和（0.26±0.07）%，在统计学上差异均具有统计学意义（均P<0.01）；模型组与对照组外周血IL-4水平分别为（48.98±3.78）和（26.44±5.64）pg·mL^-1，IL-6水平分别为（44.42±2.55）和（24.21±2.25）pg·mL^-1，IL-10水平分别为（37.67±5.34）和（19.06±3.76）pg·mL^-1，TNF-α水平分别为（79.11±6.32）和（35.78±13.17）pg·mL^-1，IL-12水平分别为（28.28±2.33）和（55.39±3.47）ng·mL^-1，IFN-γ水平分别（238.67±35.78）和（517.72±52.15）pg·mL^-1，在统计学上差异均具有统计学意义（均P<0.01）。解毒地黄汤组低、中、高剂量组及阳性药物组NLRP3 mRNA表达水平分别为1.75±0.10、1.59±0.11、1.31±0.11和1.48±0.10，ASC mRNA表达水平分别为2.69±0.10、1.96±0.14、1.54±0.18和1.89±0.09，Caspase-1 mRNA表达水平分别为2.85±0.08、2.24±0.26、1.58±0.22和1.94±0.16，除解毒地黄汤组低剂量外，各组与模型组在统计学差异均具有统计学意义（均P<0.05、P<0.01）；PINK1平均荧光强度分别为87.50±9.38、98.75±7.13、107.28±6.60和94.75±7.11，Parkin平均荧光强度分别为94.11±8.17、103.13±8.59、111.27±9.66和101.27±9.21，Beclin1平均荧光强度分别为100.2±7.11、107.92±6.84、122.65±8.36和105.44±7.55，除解毒地黄汤组低剂量外，各组与模型组在统计学差异均具有统计学意义（P<0.05、P<0.01）；腹腔液Th1占比分别为（1.34±0.13）、（1.78±0.21）、（2.89±0.23）和（2.47±0.24）%；腹腔液Th2占比分别为（1.65±0.18）、（1.06±0.11）、（0.57±0.14）和（0.68±0.07）%，除解毒地黄汤组低剂量外，各组与模型组在统计学差异均具有统计学意义（均P<0.05、P<0.01）；外周血IL-4水平分别为（41.67±4.31）、（36.39±5.93）、（33.61±4.30）和（33.94±5.81）pg·mL^-1；IL-6水平分别为（39.10±2.89）、（32.85±3.98）、（28.91±1.91）和（27.63±1.98）pg·mL^-1；IL-10水平分别为（32.11±7.50）、（26.28±4.65）、（23.22±4.35）和（26.22±6.15）pg·mL^-1；TNF-α水平分别为（64.13±13.12）、（57.44±13.62）、（47.45±10.06）和（50.78±10.9）pg·mL^-1；IL-12水平分别为（34.09±2.49）、（38.53±3.84）、（42.67±3.87）和（41.93±4.39）ng·mL^-1；IFN-γ水平分别为（286.57±54.77）、（358.59±52.15）、（435.28±73.12）和（405.33±39.33）pg·mL^-1，除解毒地黄汤组低剂量外，各组与模型组在统计学差异均具有统计学意义（P<0.05、P<0.01）。

结论

解毒地黄汤可以改善结肠癌的炎症微环境，这可能与线粒体自噬调控NLRP3炎症小体有关。

关键词

解毒地黄汤 / 结直肠癌 / NLRP3炎症小体 / 线粒体自噬

Abstract

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Objective

To explore the intervention effect of jiedu dihuang decoction on the inflammatory microenvironment of colon cancer by regulating mitophagy and nucleotide - binding oligomerization domain (NOD) like receptor hot protein domain-related protein 3 (NLRP3) inflammasome.

Methods

The animal model was successfully replicated by intraperitoneal injection of azoxymethane (AOM) combined with feeding dextran sodium sulfate (DSS). The animals were randomly divided into A: control group, B: model group, C, D, E: Jiedu Dihuang Decoction low, medium and high dose group, F: positive drug group. Ninety male C57BL/6 mice were randomly divided into 6 groups of 15 mice each, and all groups were treated continuously for 10 weeks. The pathological changes of rectal tissue were evaluated by hematoxylin-eosin (HE) staining. Immunofluorescence staining was used to observe the expression site and intensity of PTEN-induced kinase 1 (PINK1), Parkinson’s disease protein (Parkin) and B-cell lymphoma-2-interacting myosin-like coiled-coil protein 1 (Beclin1) in colon tissue. The expression of NLRP3, apoptosis-associated speck-like protein (ASC) and Caspase-1 mRNA in rectum was detected by polymerase chain reaction (PCR). The protein expression levels of PINK1, Parkin, Beclin1, NLRP3, ASC and Caspase-1 were detected by Western blotting (WB). The ratio of helper T cell 1 (Th1) to helper T cell 2 (Th2) was detected by flow cytometry. The levels of interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-12 (IL-12), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) were detected by enzyme-linked immunosorbent assay (Elisa).

Results

The mRNA expression levels of NLRP3, ASC and Caspase-1 in the model group were 2.06±0.22, 3.03±0.16 and 3.28±0.22, respectively, while those in the control group were 1.01±0.17, 1.01±0.19 and 1.00±0.11, respectively, with statistically significant differences between the two groups (all P<0.01). The mean fluorescence intensities of PINK1, Parkin and Beclin1 in the model group were 76.39±4.89, 83.12±5.01 and 87.43±3.77, respectively, compared with 122.99±6.93, 138.38±3.84 and 143.22±9.42 in the control group, and the differences were statistically significant (all P<0.01). As for the proportion of T helper cells in peritoneal fluid, the model group showed a significantly lower Th1 proportion (0.74±0.09)% and a significantly higher Th2 proportion (2.05±0.26)% compared with the control group [(4.78±0.62)% and (0.26±0.07)%, respectively, all P<0.01]. In terms of peripheral blood cytokine levels, the concentrations of IL-4, IL-6, IL-10 and TNF-α in the model group were (48.98±3.78) pg·mL^-1, (44.42±2.55) pg·mL^-1, (37.67±5.34) pg·mL^-1 and (79.11±6.32) pg·mL^-1, respectively, which were significantly higher than those in the control group [(26.44±5.64) pg·mL^-1, (24.21±2.25) pg·mL^-1, (19.06±3.76) pg·mL^-1 and (35.78±13.17) pg·mL^-1, respectively]. Conversely, the levels of IL-12 and IFN-γ in the model group were (28.28±2.33) ng·mL^-1 and (238.67±35.78) pg·mL^-1, respectively, which were significantly lower than those in the control group [(55.39±3.47) ng·mL^-1 and (517.72±52.15) pg·mL^-1, respectively], with all differences reaching statistical significance (all P<0.01). The mRNA expression levels of NLRP3 in the low, medium, high-dose Jiedu-Dihuang Decoction groups and the positive drug group were 1.75±0.10, 1.59±0.11, 1.31±0.11 and 1.48±0.10, respectively; the mRNA expression levels of ASC were 2.69±0.10, 1.96±0.14, 1.54±0.18 and 1.89±0.09, respectively; and the mRNA expression levels of Caspase-1 were 2.85±0.08, 2.24±0.26, 1.58±0.22 and 1.94±0.16, respectively. Except for the low-dose Jiedu-Dihuang Decoction group, the differences between the other treatment groups and the model group were statistically significant (P<0.05, P<0.01). The mean fluorescence intensities of PINK1 in each treatment group were 87.50±9.38, 98.75±7.13, 107.28±6.60 and 94.75±7.11, respectively; those of Parkin were 94.11±8.17, 103.13±8.59, 111.27±9.66 and 101.27±9.21, respectively; and those of Beclin1 were 100.2±7.11, 107.92±6.84, 122.65±8.36 and 105.44±7.55, respectively. Except for the low-dose Jiedu-Dihuang Decoction group, the differences between the other treatment groups and the model group were statistically significant(P<0.05, P<0.01). The proportions of Th1 in peritoneal fluid of each treatment group were (1.34±0.13)%, (1.78±0.21)%, (2.89±0.23)% and (2.47±0.24)%, respectively; and the proportions of Th2 were (1.65±0.18)%, (1.06±0.11)%, (0.57±0.14)% and (0.68±0.07)%, respectively. Except for the low-dose Jiedu-Dihuang Decoction group, the differences between the other treatment groups and the model group were statistically significant (all P<0.05, P<0.01). The levels of IL-4 in peripheral blood of each treatment group were (41.67±4.31), (36.39±5.93), (33.61±4.30) and (33.94±5.81) pg·mL^-1, respectively; those of IL-6 were (39.10±2.89), (32.85±3.98), (28.91±1.91) and (27.63±1.98) pg·mL^-1, respectively; those of IL-10 were (32.11±7.50), (26.28±4.65), (23.22±4.35) and (26.22±6.15) pg·mL^-1, respectively; those of TNF-α were (64.13±13.12), (57.44±13.62), (47.45±10.06) and (50.78±10.9) pg·mL^-1, respectively; those of IL-12 were (34.09±2.49), (38.53±3.84), (42.67±3.87) and (41.93±4.39) ng·mL^-1, respectively; and those of IFN-γ were (286.57±54.77), (358.59±52.15), (435.28±73.12) and (405.33±39.33) pg·mL^-1, respectively. Except for the low-dose Jiedu-Dihuang Decoction group, the differences between the other treatment groups and the model group were statistically significant(all P<0.05, P<0.01)

Conclusion

Jiedu-Dihuang Decoction can improve the inflammatory microenvironment of colon cancer, which may be related to the regulation of NLRP3 inflammasome by mitochondrial autophagy.

Key words

Jiedu Dihuang Decoction / colon cancer / NLRP3 inflammasome / mitochondrial autophagy

引用本文

李育民, 黄灿灿, 归明彬, 岳斌, 李兆桓, 王芸, 薛秀芳, 裴正学. 解毒地黄汤拮抗结肠癌线粒体自噬及NLRP3活化的作用及机制研究. 中国临床药理学杂志, 2026 , 42 (3) : 356 -363 . DOI: 10.13699/j.cnki.1001-6821.2026.03.010

Yu-min LI, Can-can HUANG, Ming-bin GUI, Bin YUE, Zhao-huan LI, Yun WANG, Xiu-fang XUE, Zheng-xue PEI. Research on the effect and mechanism of Jiedu Dihuang Decoction in antagonizing mitochondrial autophagy and NLRP3 activation in colorectal cancer[J]. Chinese Journal of Clinical Pharmacology, 2026 , 42 (3) : 356 -363 . DOI: 10.13699/j.cnki.1001-6821.2026.03.010

基金

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甘肃省中医药管理局基金资助项目(GZKP-2023-23)
甘肃省科技计划基金资助项目(25JRRA429)
兰州市科技计划基金资助项目(2024-9-163)

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2026年第42卷第3期

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文章信息

doi: 10.13699/j.cnki.1001-6821.2026.03.010

接收时间：2025-09-29
首发时间：2026-08-06
出版时间：2026-02-17

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收稿日期：2025-09-29

基金

甘肃省中医药管理局基金资助项目(GZKP-2023-23)

甘肃省科技计划基金资助项目(25JRRA429)

兰州市科技计划基金资助项目(2024-9-163)

作者信息

^1.甘肃省第二人民医院/西北民族大学附属医院肿瘤内科甘肃兰州 730030

^2.南通市中医院妇科，江苏南通 226000

^3.中国人民解放军联勤保障部队第九四〇医院结直肠肛门外科甘肃兰州 730050

^4.甘肃中医药大学中医临床学院，甘肃兰州 730000

^5.甘肃省肿瘤医院中西医结合科，甘肃兰州 730050

通讯作者:

薛秀芳，副研究员 MP: 13919770279 E-mail: 243825948@qq.com

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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