动物造模,药效评价,前列腺增生 z-bio 2024-11-18 381

　　1. Modeling material animals: Adult male inbred BALB/c mice, inbred BALB/c mice at 16-18 days of gestation; Medications: Urinary reproductive sinus stroma, penicillin, streptomycin, D-Hank's solution, trypsin, calf serum culture medium, ether.

　　2. Preparation of UGM modeling method: Inbred BALB/c mice at 16-18 days of gestation were euthanized by cervical dissection, and the embryos were removed. The embryos were then dissected under a stereomicroscope in a 0-4 ℃ D-Hank solution containing penicillin and streptomycin, and the embryonic bladder, Wolffian and Mullerian tubes were trimmed, leaving behind a complete urogenital sinus (UGS). Place UGS in a 1% trypsin solution at 4 ℃ and digest for 120 minutes to remove epithelial components and retain UGM. Wash with a culture medium containing 50% calf serum to terminate trypsin digestion. The prepared UGM should be stored in D-Hank culture medium and used within 3 hours. The entire operation process is carried out under sterile conditions.

　　Animal inoculation: Animals were randomly divided into: normal control group, sham surgery group, and UGM implantation group. Inhale ether anesthesia and properly fix the mouse on the operating table. Make a midline incision to expose the bladder and ventral prostate, and implant the prepared UGM into the ventral prostate. The sham surgery group did not implant UGM, and the remaining steps were the same as above.

　　3. The principle of modeling is to induce benign prostatic hyperplasia in mice using UGM from mouse embryos, and establish an animal model of benign prostatic hyperplasia.

　　4. After modeling, the UGM implantation group showed a significant increase in prostate wet weight compared to the control and sham surgery groups. The pathological manifestations of the control and sham surgery groups were basically the same, with a single-layer low columnar glandular epithelium and smaller papillary protrusions. The gland appears tubular, with a small amount of secretion inside the gland cavity and less fibrous stroma. The UGM implantation group showed glandular and fibrous tissue hyperplasia, with glandular hyperplasia being the main type. The glandular cavity was round, and the glandular epithelium was high columnar with hyperplasia, resulting in the formation of more papillae protruding into the cavity. There was an increase in secretion in the glandular cavity and fibrous stroma hyperplasia.

　　5. Precautions: To verify that the prepared UGM meets the requirements of this experiment, the UGM will be studied as follows: observe whether there are residual Wolffian and Mullerian tubes and other accessory structures under a surgical microscope, continuously slice the UGM, and observe whether there are residual epithelial components by HE staining. To further demonstrate that the digested UGM has no epithelial components and is active, UGM was implanted under the renal capsule of male mice of the same lineage. Four weeks later, the graft was cut and continuously sliced for HE staining observation.