Establishment of Preeclampsia Model in Goat and Evaluation on Maternal Biological Characteristics

doi:10.12300/j.issn.1674-5817.2023.036

Abstract

Abstract:

Objective Establish an animal model of preeclampsia in goats, collect data on various physiological indicators and maternal biological characteristics of the disease model to compare with clinical feature of the disease in humans, provide reference for the prevention and treatment of preeclampsia in humans. Methods Twenty-three goats bred in Chongming district were divided into three groups: Control group, no surgical procedure was performed on animals of this group; sham group, ewes in this group underwent the sham operation on the 100±5th day of gestation, and only the uterine artery was exposed and dissociated; surgical group, a silver vascular clamp was clipped on one side of the uterine body artery of the ewe to narrow the inner diameter of the artery at the same gestation period (100±5) days. Heart rate and hindlimb blood pressure were continuously monitored in control and surgical ewes from 100 to 140 d of gestation, and blood flow data within the lateral branches of the uterine arteries of ewes in the sham group were collected using a hemodynamometer in combination with a hemodynamic probe and an animal physiological signal collector, as well as changes in blood flow within the uterine arteries in the lateral branches of the uterine arteries of the surgical group before and after placement of vascular clips in the surgical ewes. At the expected date of delivery, jugular vein blood was taken from ewes for routine blood test, creatinine, urea nitrogen, and blood ion analysis; urine was also collected from ewes in each group for analysis of urinary protein and urinary creatinine. All experimental groups were subjected to cesarean section on the 140±5th day of gestation in ewes, and then liver, kidney, uterus and placenta tissues were taken from ewes in each group and stained with HE for pathological observation. Results After 15 minutes of preeclampsia modeling surgery, blood flow volume remained stable in the vessel stenosis segment and the volume differential was relatively reduced in comparison to the control group and sham group (P＜0.05，P＜0.01). Compared with the control group, the ewes in the surgical group showed prenatal changes such as increased serum osmolality, decreased hemoglobin, increased blood glucose and urea nitrogen values, as well as increased levels of calcium, sodium, and chloride ions (all P<0.05) and proteinuria, with urinary creatinine and urinary protein-creatinine ratios were significantly higher than those in the control group and sham group (all P<0.05). The elastic lamina of the uterine body arteries on the operated side of the animals in the surgical group was thicker than that on the opposite side, but the structure was loose. The placenta on the operated side showed pathological changes such as cell interstitial swelling and inflammatory cell infiltration. The above physiological index characteristics were more consistent with the clinical features of human preeclampsia disease. Conclusion In this experiment, we successfully constructed a goat preeclampsia model and obtained data on relevant physiological indexes of this model, which further verified the correlation between preeclampsia disease and uterine artery lesions.

Key words: Preeclampsia, Animal model, Blood flow analysis, Biological characteristics, Goat

CLC Number:

R-332

Jin LU, Jian WANG, Lian ZHU, Guofeng YAN, Zhengwen MA, Yao LI, Jianjun DAI, Yinqiu ZHU, Jing ZHOU. Establishment of Preeclampsia Model in Goat and Evaluation on Maternal Biological Characteristics[J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 371-380.

Figures/Tables 6

Figure 1 Anterior exposure of uterine arteries and the position of the silver clamp placement on uterine body arteryNote：A， Anterior exposure of uterine arteries （UA， uterine artery；UB， uterine body；R， right uterine body artery；L， left uterine body artery；SC， placement of silver clamp）； B， Uterine body artery been partially closed by silver clamp.

Figure 2 Blood flow， blood pressure and heart rate of pregnant goats after preeclampsia modellingNote：A， Blood flow changes of uterine body artery； B， Changes of hind limb diastolic blood pressure；C， Changes of hind limb pulse pressure； D， Change of heart rate （BPM， beat per min）. In control group （n=11）， no surgical operation was done on animals； in sham group （n=4）， sham surgery was performed on the 100th day of gestation， to be specific， only their uterine body arteries were exposed and freed； in surgical group （n=8）， preeclampsia modeling was performed by placing silver vascular clamps on one of the uterine body arteries of the animals on the 100th day of gestation to close their internal diameters partially. For comparison between groups， *P<0.05， **P<0.01， ***P<0.001， ****P<0.000 1.

Table 1 Comparison of blood routine between groups in preeclampsia goat model

项目 Item	手术组（n=8） Surgical group (n=8)		对照组（n=11） Control group (n=11)
项目 Item	术后5 d Postoperation 5 d	产前 Before parturition	对照组（n=11） Control group (n=11)
白细胞计数/(10³·μL^-1) White blood cell count/(10³·μL^-1)	38.26±2.81	34.00±2.99	40.62±1.70
淋巴细胞百分比/% Lymphocyte/%	58.68±2.28	58.42±4.22	60.94±2.05
中间细胞百分比/% Intermediate cell/%	7.20±1.07	7.60±1.09	6.61±0.73
粒细胞百分比/% Granulocyte/%	34.12±1.77	33.98±3.18	32.45±1.63
淋巴细胞计数/(10³·μL^-1) Lymphocyte count/(10³·μL^-1)	22.50±1.86	20.13±2.85	24.81±1.44
粒细胞计数/(10³·μL^-1) Granulocyte count/(10³·μL^-1)	12.90±0.81	11.30±0.64	13.06±0.68
红细胞计数/(10⁶·μL^-1) Red blood cell count/(10⁶·μL^-1)	1.14±0.14	0.65±0.04^*	1.58±0.17
血红蛋白计数/(g·L^-1) haemoglobin count/(g·L^-1)	101.40±4.24	87.75±4.71^*	117.27±5.32
血细胞压积/% Packed cell volume/%	4.52±0.62	2.50±0.16^*	6.49±0.76
平均红细胞压积/fL Mean corpuscular volume/fL	40.18±0.31	39.53±0.30^*	41.16±0.30
平均红细胞血红蛋白量/pg Mean corpuscular hemoglobin/pg	94.00±10.30	135.18±6.02^****	77.99±4.33
平均红细胞血红蛋白浓度/(g·L^-1) Mean corpusular hemoglobin concerntration/(g·L^-1)	2 397±286	3 526±154^****	1 924±118
红细胞分别宽度变异系数/% coefficient variation of red blood cell volume distribution width/%	17.26±0.61	16.33±0.69	17.41±0.43
中间细胞计数/(10³·μL^-1) Intermediate cell count/(10³·μL^-1)	2.58±0.44	2.40±0.22	2.99±0.34

Figure 3 Blood and urinary biochemical results of preeclampsia modeling goat before parturitionNote：A， Blood glucose； B， Renal functional indicators； C， Serum iron indicators （PHOS， phosphate； Ca， calcium； Na， Sodium； K， potassium； Na/K， sodium potassium ratio； Cl， chlorine； OSM CALC， osmotic pressure）； D， Urinary creatinine and urinary total protein； E， Creatinine/protein ratio. In the control group （n=11）， no surgical operation was done on animals； in sham group （n=4）， sham surgery was performed on the 100th day of gestation，to be specific， only their uterine body arteries were exposed and freed；and in surgical group （n=8）， preeclampsia modeling was performed by placing silver vascular clamps on one of the uterine body arteries of the animals on the 100th day of gestation to close some of their internal diameters. Comparison between groups， *P<0.05， **P<0.01， ***P<0.001， ****P<0.000 1.

Figure 4 Pathologic changes in the placenta on the operated and non-operated side during parturition in goats after preeclampsia modeling（HE staining）Note：A， Normal placenta of ewes in control group （×1.1， the blue squares are the sampling sites for figure B and C ）； B， Trophoblasts of normal placenta in control group （×16.9）； C， Villous stroma of normal placenta in control group （×27.2）； D， Surgical side placenta of ewes in surgical group （×1.1， the blue squares are the sampling sites for figure E and F ）； E， Trophoblasts of surgical side placenta of ewes in surgical group （×28.7）； F， Villous stroma of surgical side placenta of ewes in surgical group （×34.3）. Scale bars of figure A and D are 1 000 μm. Proportional scales of figure B， C， E and F are 50 μm.

Figure 5 Comparison of the appearance and pathology of the uterine body arteries on surgical and non-surgical side of the uterus at the time of parturition in pregnant goats after preeclampsia modelingNote：A， Schematic diagram of the uterus and uterine arteries of ewes on the 140th day of gestation （silver clips were placed on one side of the uterine body arteries）； B-D， HE-stained histopathologic sections of the uterine body arteries （B， uterine arteries of the nonsurgical side； C， uterine arteries proximal to the surgical side； E， uterine arteries distal from the surgical side）. The scale bars figure B-D are 50 μm.

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