目的 探讨三子颗粒通过降低微小核糖核酸-205-5p（miR-205-5p）水平抑制小鼠脾虚型肠道腺瘤生长的作用。方法 取70只4周龄的雄性C57BL/6J小鼠，采用对氧化偶氮甲烷（AOM）/葡聚糖硫酸钠（DSS）诱导小鼠结直肠腺瘤模型，将建模小鼠随机分为模型组、阿司匹林（200 mg·kg^-1）组、miR inhibitor-NC （2 mg·kg^-1）组、miR-205-5p inhibitor （2 mg·kg^-1）组和三子颗粒低、中、高剂量（1.7、3.4、6.8 g·kg^-1）组，造模期间ig给药，每天1次。比较各组小鼠肠道腺瘤的数量并测量腺瘤体积；苏木素-伊红（HE）染色观察小鼠肠道腺瘤的病理情况；CCK-8法检测各组小鼠肠道腺瘤细胞增殖活力；原位末端标记（TUNEL）法检测小鼠肠道腺瘤细胞凋亡；实时荧光定量PCR（qRT-PCR）法检测肠道腺瘤miR-205-5p表达量及磷酸酯酶与张力蛋白同源物（PTEN）、B淋巴细胞瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）、Ki67 mRNA表达量；Western blotting法检测PTEN、Bcl-2、Bax、Ki67蛋白表达水平；荧光素酶活性实验验证miR-205-5p和PTEN的靶向关系。结果 与模型组比较，阿司匹林组和三子颗粒低、中、高剂量组小鼠肠道腺瘤数量、体积及细胞增殖活性均显著降低，凋亡率显著升高（P＜0.05），miR-205-5p表达量、Bcl-2、Ki67 mRNA及蛋白表达量显著降低，PTEN、Bax mRNA及蛋白表达量显著升高（P＜0.05），其中三子颗粒作用呈剂量相关性；与miR inhibitor-NC组比较，miR-205-5p inhibitor组小鼠肠道腺瘤数量、体积及细胞增殖活性均显著降低，凋亡率显著升高（P＜0.05），miR-205-5p表达量、Bcl-2、Ki67 mRNA及蛋白表达量显著降低，PTEN、Bax mRNA及蛋白表达量显著升高（P＜0.05）；荧光素酶活性实验证实miR-205-5p可靶向调控PTEN。结论 三子颗粒可抑制小鼠脾虚型肠道腺瘤生长，可能是通过下调miR-205-5p，上调PTEN、Bax表达，下调Bcl-2、Ki67表达发挥作用的。

Objective To investigate the effect of Sanzi Granules on inhibition of miR-205-5p on the growth of intestinal adenoma of spleen deficiency type in mice. Methods Seventy 4-week-old male C57BL/6J mice were treated with p-azomethane (AOM)/sodium glucan sulfate (DSS) to induce colorectal adenoma, and randomly divided the modeled mice into model group, aspirin (200 mg·kg^-1) group, miR inhibitor NC (2 mg·kg^-1) group, miR-205-5p inhibitor (2 mg·kg^-1) group, and Sanzi Granules low, medium, and high dose (1.7, 3.4, 6.8 g·kg^-1) groups. The number and volume of intestinal adenomas were measured. Hematoxylin eosin (HE) staining was used to observe the pathology of intestinal adenoma in mice. Cell counting kit - 8 (CCK-8) was used to detect the proliferation of intestinal adenoma cells of mice in each group. Detection of apoptosis of intestinal adenoma cells in mice by TdT-mediated dUTP nick end labeling (TUNEL). Real time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-205-5p in intestinal adenoma and the mRNA expression of phosphatase and tensin homolog (PTEN), B lymphoblastoma 2 (Bcl-2), Bcl-2 related X protein (Bax), Ki67. Western blotting was used to detect the protein expressions of PTEN, Bcl-2, Bax and Ki67. The targeting relationship between miR-205-5p and PTEN was verified by luciferase activity experiment. Results Compared with the model group, the number, volume and cell proliferation activity of intestinal adenomas in the aspirin group and the Sanzi Granules low, medium and high dose groups decreased, the apoptosis rate increased (P < 0.05), the expression of miR-205-5p, Bcl-2, Ki67 mRNA and protein decreased, and the expression of PTEN, Bax mRNA and protein increased (P < 0.05). The effect of Sanzi Granules was dose dependent. Compared with miR inhibitor NC group, the number, volume and cell proliferation activity of intestinal adenoma of mice in miR-205-5p inhibitor group decreased, the apoptosis rate increased (P < 0.05), the expression of miR-205-5p, Bcl-2, Ki67 mRNA and protein decreased, and the expression of PTEN, Bax mRNA and protein increased (P < 0.05). Luciferase activity experiment confirmed that miR-205-5p could target and regulate PTEN.Conclusion Sanzi Granules can inhibit the growth of intestinal adenoma of spleen deficiency type in mice, possibly by mediating miR-205-5p, down regulating the expression of Bcl-2 and Ki67, and up regulating the expression of PTEN and Bax.

R285.5

江苏省中医药管理局科技项目（JD2019SZXYB15）