Study on Sanzi Granules downregulated miR-205-5p inhibiting growth of intestinal adenoma of spleen deficiency type in mice

Abstract

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.