目的 探討三子顆粒通過降低微小核糖核酸-205-5p（miR-205-5p）水平抑制小鼠脾虛型腸道腺瘤生長的作用。方法 取70只4周齡的雄性C57BL/6J小鼠，采用對(duì)氧化偶氮甲烷（AOM）/葡聚糖硫酸鈉（DSS）誘導(dǎo)小鼠結(jié)直腸腺瘤模型，將建模小鼠隨機(jī)分為模型組、阿司匹林（200 mg·kg^-1）組、miR inhibitor-NC （2 mg·kg^-1）組、miR-205-5p inhibitor （2 mg·kg^-1）組和三子顆粒低、中、高劑量（1.7、3.4、6.8 g·kg^-1）組，造模期間ig給藥，每天1次。比較各組小鼠腸道腺瘤的數(shù)量并測量腺瘤體積；蘇木素-伊紅（HE）染色觀察小鼠腸道腺瘤的病理情況；CCK-8法檢測各組小鼠腸道腺瘤細(xì)胞增殖活力；原位末端標(biāo)記（TUNEL）法檢測小鼠腸道腺瘤細(xì)胞凋亡；實(shí)時(shí)熒光定量PCR（qRT-PCR）法檢測腸道腺瘤miR-205-5p表達(dá)量及磷酸酯酶與張力蛋白同源物（PTEN）、B淋巴細(xì)胞瘤-2（Bcl-2）、Bcl-2相關(guān)X蛋白（Bax）、Ki67 mRNA表達(dá)量；Western blotting法檢測PTEN、Bcl-2、Bax、Ki67蛋白表達(dá)水平；熒光素酶活性實(shí)驗(yàn)驗(yàn)證miR-205-5p和PTEN的靶向關(guān)系。結(jié)果 與模型組比較，阿司匹林組和三子顆粒低、中、高劑量組小鼠腸道腺瘤數(shù)量、體積及細(xì)胞增殖活性均顯著降低，凋亡率顯著升高（P＜0.05），miR-205-5p表達(dá)量、Bcl-2、Ki67 mRNA及蛋白表達(dá)量顯著降低，PTEN、Bax mRNA及蛋白表達(dá)量顯著升高（P＜0.05），其中三子顆粒作用呈劑量相關(guān)性；與miR inhibitor-NC組比較，miR-205-5p inhibitor組小鼠腸道腺瘤數(shù)量、體積及細(xì)胞增殖活性均顯著降低，凋亡率顯著升高（P＜0.05），miR-205-5p表達(dá)量、Bcl-2、Ki67 mRNA及蛋白表達(dá)量顯著降低，PTEN、Bax mRNA及蛋白表達(dá)量顯著升高（P＜0.05）；熒光素酶活性實(shí)驗(yàn)證實(shí)miR-205-5p可靶向調(diào)控PTEN。結(jié)論 三子顆?？梢种菩∈笃⑻撔湍c道腺瘤生長，可能是通過下調(diào)miR-205-5p，上調(diào)PTEN、Bax表達(dá)，下調(diào)Bcl-2、Ki67表達(dá)發(fā)揮作用的。

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

江蘇省中醫(yī)藥管理局科技項(xiàng)目（JD2019SZXYB15）