补中益气加味方对裸鼠结直肠癌移植瘤Klotho蛋白及Caspase-1、IL-1β、IL-18表达的影响*

作者：王 伟1，周望潮2，谢 彪1

单位：1.湖南中医药大学第二附属医院，湖南 长沙 410005； 2.湖南中医药大学，湖南 长沙 410208

引用：引用：王伟，周望潮，谢彪.补中益气加味方对裸鼠结直肠癌移植瘤Klotho蛋白及Caspase-1、IL-1β、IL-18表达的影响[J].中医药导报,2025,31(12):14-19.

DOI：10.13862/j.cn43-1446/r.2025.12.003

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摘要：

目的：探讨补中益气加味方通过调控Klotho蛋白及细胞焦亡抑制结直肠癌移植瘤生长的作用机制。方法：建立HCT116人结直肠癌细胞裸鼠移植瘤模型，15 d成模后随机分为模型组、氟尿嘧啶（5-FU）组、补中益气加味方组、Klotho组、补中益气加味方+Klotho组，分别给予生理盐水、5-FU（25 mg/kg）、补中益气加味方（53.43 g/kg）、外源性Klotho蛋白（0.01 mg/kg）和联合使用补中益气加味方及外源性Klotho蛋白。连续干预12 d，在第0天、4天、8天、12 天时测定瘤体体积，第12天测定瘤体质量，HE染色观察组织病理，免疫组化法检测肿瘤组织中Klotho、胱天蛋白酶1-（Caspase-1）、白细胞介素-1β（IL-1β）和IL-18的表达水平。结果：与模型组相比，5-FU组、补中益气加味方组、Klotho组、补中益气加味方+Klotho组的裸鼠移植瘤质量明显降低（P<0.01或P<0.05）。在药物治疗后的第4天、第8天和第12天，5-FU组、补中益气加味方组、Klotho组和补中益气加味方+Klotho组的裸鼠移植瘤体积明显小于模型组（P<0.01），各治疗组治疗后的移植瘤体积增大幅度均低于模型组。HE染色显示治疗后切片可见细胞核固缩，胞质空泡化，细胞排列疏松，间质胶原纤维增生。免疫组化结果显示5-FU组、补中益气加味方组、Klotho组和补中益气加味方+Klotho组的裸鼠移植瘤Klotho表达高于模型组（P<0.01或P<0.05），补中益气加味方组、Klotho组和补中益气加味方+Klotho组的Caspase-1、IL-1β和IL-18表达低于模型组（P<0.01或P<0.05）。结论：补中益气加味方可有效抑制裸鼠结直肠癌移植瘤生长，其机制可能与上调Klotho表达、抑制Caspase-1依赖性细胞焦亡及降低促炎因子IL-1β和IL-18的释放有关。

关键词：结直肠癌；补中益气加味方；Klotho；细胞焦亡；胱天蛋白酶-1；白细胞介素-1β；白细胞介素-18；裸鼠

Abstract：

Objective: To explore the mechanism of action of the modified Buzhong Yiqi formula in inhibiting the growth of colorectal cancer xenografts by regulating Klotho protein and pyroptosis. Methods: A nude mouse xenograft model was established using HCT116 human colorectal cancer cells. After 15 days of model establishment, the mice were randomly divided into model group, fluorouracil group (5-FU group), modified Buzhong Yiqi formula group, Klotho group, and modified Buzhong Yiqi formula + Klotho group. The treatments administered included normal saline, 5-FU (25 mg/kg), modified Buzhong Yiqi formula (53.43 g/kg), exogenous Klotho (0.01 mg/kg), and a combination of modified Buzhong Yiqi formula and exogenous Klotho. Continuous intervention lasted for 12 days. Tumor volume was measured on day 0, day 4, day 8 and day 12. The tumor mass was measured on the 12th day, and tissue pathology through was observed by HE staining. The expression levels of Klotho, Caspase-1, IL-1β, and IL-18 in tumor tissue were assessed using immunohistochemistry. Results: Compared to the model group, the tumor masses of nude mice decreased significantly in the 5-FU group, modified Buzhong Yiqi formula group, Klotho group, and modified Buzhong Yiqi formula + Klotho group (P<0.01 or P<0.05). On the 4th, 8th and 12th days post-treatment, the 5-FU group, modified Buzhong Yiqi formula group, Klotho group, and modified Buzhong Yiqi formula + Klotho group showed lower tumor volumes than model group (P<0.01). The increase in transplanted tumor volume after treatment was lower in all treatment groups than in the model group. HE staining showed that in the post-treatment sections, there was nuclear shrinkage, cytoplasmic vacuolation, loose cell arrangement, and increased collagen fibers in the interstitium. Immunohistochemical results demonstrated that the 5-FU group, modified Buzhong Yiqi formula group, Klotho group, and modified Buzhong Yiqi formula + Klotho group showed higher expression of Klotho than model group (P<0.01 or P<0.05), and that the modified Buzhong Yiqi formula group, Klotho group, and modified Buzhong Yiqi formula + Klotho group showed lower expressions of Caspase-1, IL-1β and IL-18 than model group (P<0.01 or P<0.05). Conclusion: The modified Buzhong Yiqi formula can effectively inhibit the growth of colorectal cancer xenografts in nude mice. Its mechanism may be related to the up-regulation of Klotho expression, inhibition of Caspase-1-dependent pyroptosis, and reduction in the release of pro-inflammatory factors (IL-1β and IL-18).

Key words：colorectal cancer; modified Buzhong Yiqi formula; Klotho; pyroptosis of cells; Caspase-1; IL-1β; IL-18; nude mice

发布时间：2025-12-31

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