Rat Model of A2M Overexpression

SPF Sprague–Dawley rats (6~8 weeks age, 180~200 g weight) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (SCXK 2012-0001, Beijing, China). Based on a previous report [25 (link)], we established an A2M-overexpression rat model via tail vein injection as previously described [26 (link)]. Briefly, on gestational day (GD) 8.5, rats (excluding non-pregnant rats) were injected with adenoviruses expressing A2M (OBiO Technology Co., Shanghai, China), and the sequencing results are shown in Additional file 1: Supplementary Result 1. We injected an adenoviral dose of approximately 1–2×10⁹ pfu per animal, and the adenoviruses were dissolved in phosphate-buffered saline (PBS) to a total volume of 400 μl. The treated rats were sacrificed on GD19.5 (corresponding to the third trimester) for further study. The following parameters were assessed: blood pressure, blood flow, and proteinuria. The primary outcome of this study will be hypertension with blood pressure measurement. Secondary outcomes constitute blood flow, proteinuria, and histological analyses to measure the morphology and cell function of the spiral artery and placental vascular. For the rat samples, the pregnant rats were first euthanized to collect placentas and fetuses. According to Resource Equation Approach [27 (link)], a total of 20 rats were studied, the rats were randomly divided into two groups (n = 10 in each group): the control and A2M-overexpression groups (note: the rats used in this experiment came from at least three different modelling batches). Random numbers were generated using the standard = RAND() function in Microsoft Excel. Each rat was euthanized by cervical dislocation after the experiment. Experiments involving animals were performed in accordance with the ARRIVE guidelines. All experimental processes involving animal treatments were conducted in accordance with the procedures of the Ethics Committee for Animal Experimentation, Jinan University (approval number: 20210302-46).