Screening of Hypoxia-Sensitive and Hypoxia-Tolerant Wistar Rats and Preliminary Exploration of Hypoxia Sensitivity in Their G1 Generation

doi:10.12300/j.issn.1674-5817.2024.017

Abstract

Abstract:

Objective By screening and passaging G₀ generation Wistar rats, we obtained hypoxia-sensitive and hypoxia-tolerant G₁ generation rats, and then the differences in hypoxia sensitivity among these rats were preliminarily explored. Methods 200 Wistar rats (half male and half female) were selected as G₀ generation and placed in a controlled oxygen concentration system. The hypoxia tolerance time, which refers to the time from placement to near death, was recorded for the G₀ generation rats at an oxygen volume fraction of 3%. 30 rats (half male and half female) with the shortest hypoxia tolerance time were selected for mating and passage to obtain G₁ generation hypoxia-sensitive rats. Similarly, 30 rats (half male and half female) with the longest hypoxia tolerance time were selected for mating and passage to obtain G₁ generation hypoxia-tolerant rats. An additional 24 standard Wistar rats were randomly divided into two groups: a control group and a model group, with 12 rats in each group (half male and half female). The control group was kept in a normoxic environment, while the model group, along with the G₁ generation hypoxia-sensitive rats (G₁sensitive group) and G₁ generation hypoxia-tolerant rats (G₁ tolerant group), were placed in a hypobaric hypoxia chamber (simulating an altitude of 5 000 m). After 12 hours, various indicators, including blood gas, complete blood count, blood biochemistry, pathological sections, and hypoxia-related genes were detected or observed to compare the differences in hypoxia sensitivity among the 4 groups. Results Compared with the G₀ generation standard rats, the hypoxia tolerance time of G₁ generation rats was significantly prolonged (P<0.01). Compared with the model group, the oxygen saturation (SatO₂) in G₁ tolerant group was significantly higher (P<0.05). In the G₁ sensitive group, the levels of white blood cell (WBC) count, neutrophil (NEUT) count, hemoglobin (HGB) concentration, hematocrit (HCT), red blood cell distribution width (RDW), platelet (PLT), and creatinine (Cr) significantly increased (P<0.05 or P<0.01), while actual bicarbonate (AB) content significantly decreased (P<0.05), and the brain and lung coefficients were significantly elevated (P<0.05). In addition, pathological section results showed that the brain and lung tissues in the model group, G₁ sensitive group, and G₁ tolerant group all suffered from significant damage, with no evident differences in the gene expression levels of hypoxia-inducible factor-1 α (HIF-1α) and vascular endothelial growth factor A (VEGFA) in brain tissues among the three groups (P>0.05). Conclusion Compared with standard rats, G₁ generation hypoxia-sensitive/tolerant rats exhibit good signs of hypoxia sensitivity/tolerance traits, but further screening and passage are still needed to purify them.

Key words: Wistar rats, Hypoxia-sensitive, Hypoxia-tolerant, Blood gas analysis, Blood indicators

CLC Number:

R-332

XIAO Pan,WANG Hongyi,LU Lu,et al. Screening of Hypoxia-Sensitive and Hypoxia-Tolerant Wistar Rats and Preliminary Exploration of Hypoxia Sensitivity in Their G₁Generation[J]. Laboratory Animal and Comparative Medicine, 2024, 44(4): 374-383. DOI: 10.12300/j.issn.1674-5817.2024.017.

Figures/Tables 6

Table 1 Hypoxia tolerance time of G0 and G1 generation rats

遗传代数及低氧敏感性 Genetic algebra and hypoxia sensitivity	雄性Male				雌性Female
遗传代数及低氧敏感性 Genetic algebra and hypoxia sensitivity	n	$x ¯$ ±s		范围Range	n	$x ¯$ ±s	范围Range
G₀代 G₀ generation	100	928.18±524.06	232～2 965		100	976.98±738.53	151～3 280
G₁代敏感型 G₁hypoxia-sensitive type	232	832.15±486.19	165～2 838		251	1 171.68±399.67^acc	355～1 921
G₁代耐受型 G₁hypoxia-tolerant type	266	1 300.59±712.44^aabbcc	295～3 121		283	1 748.69±932.60^aabbccdd	384～3 621

Table 1 Hypoxia tolerance time of G0 and G1 generation rats

遗传代数及低氧敏感性 Genetic algebra and hypoxia sensitivity	雄性Male				雌性Female
遗传代数及低氧敏感性 Genetic algebra and hypoxia sensitivity	n	$x ¯$ ±s		范围Range	n	$x ¯$ ±s	范围Range
G₀代 G₀ generation	100	928.18±524.06	232～2 965		100	976.98±738.53	151～3 280
G₁代敏感型 G₁hypoxia-sensitive type	232	832.15±486.19	165～2 838		251	1 171.68±399.67^acc	355～1 921
G₁代耐受型 G₁hypoxia-tolerant type	266	1 300.59±712.44^aabbcc	295～3 121		283	1 748.69±932.60^aabbccdd	384～3 621

Table 2 Measurement results of blood gas indicators in rats

组别 Group	酸碱度 pH	二氧化碳分压/mmHg PaCO₂/mmHg	氧分压/mmHg PaO₂/mmHg	氧饱和度/% SatO₂/%	标准碳酸氢盐c/(mmol·L^-1) SB c/(mmol·L^-1)	实际碳酸氢盐c/(mmol·L^-1) AB c/(mmol·L^-1)
空白对照组 Control group	7.36±0.05	49.65±6.85	84.05±6.26	92.50±2.07	25.80±1.23	26.10±2.23
模型对照组 Model group	7.30±0.04^a	45.43±7.04	78.29±9.59	85.29±2.69^aa	21.29±1.80^aa	21.86±1.77^aa
G₁敏感组 G₁sensitive group	7.30±0.02^a	42.57±1.99^a	76.29±3.15^a	84.00±2.16^aa	20.43±2.30^aa	19.29±2.50^aab
G₁耐受组 G₁tolerant group	7.31±0.03^a	46.86±1.07	79.43±1.81	88.86±5.34^abc	22.86±2.97^ac	21.58±11.51^aac

Table 3 Measurement results of complete blood count indicators in rats

检测指标 Detection index	空白对照组 Control group	模型对照组 Model group	G₁敏感组 G₁ sensitive group	G₁耐受组 G₁tolerant group
白细胞计数, ×10⁹/L WBC, ×10⁹/L	4.03±1.13	4.55±0.47	5.58±1.50^aab	4.33±0.46
中性粒细胞数, ×10⁹/L NEUT, ×10⁹/L	0.92±0.24	1.09±0.16	1.59±0.54^aabb	0.94±0.13^cc
淋巴细胞数, ×10⁹/L LYMPH, ×10⁹/L	2.95±0.84	3.75±1.78	3.82±0.87	3.47±1.57
中性粒细胞百分比/% NEUT%	22.99±2.20	29.81±14.58	30.79±7.38	27.31±11.20
红细胞计数, ×10¹²/L RBC, ×10¹²/L	7.77±0.31	9.09±0.64^aa	9.49±0.63^aa	8.59±0.26^aacc
血红蛋白 ρ/(g·L^-1) HGB ρ/(g·L^-1)	141.86±3.29	165.00±9.47^aa	175.71±10.00^aabb	156.00±3.02^aabcc
血细胞比容/% HCT/%	43.79±1.44	47.78±3.09^aa	50.66±3.17^aab	44.96±1.17^bcc
红细胞分布宽度百分比/% RDW/%	15.10±0.40	24.70±0.85^aa	27.29±1.44^aabb	24.70±0.85^aacc
血小板计数, ×10⁹/L PLT, ×10⁹/L	775.00±77.59	1 085.00±82.85^aa	1 183.71±64.17^aab	994.50±75.40^aabcc
血小板分布宽度/% PDW/%	15.03±0.13	14.66±0.15^aa	14.70±0.08^aa	14.66±0.15^aa

Table 4 Measurement of biochemical indicators and organ coefficients in rats

组别 Group	肌酐z/(U·L^-1) Cr z/(U·L^-1)	天冬氨酸转氨酶 z/(U·L^-1) AST z/(U·L^-1)	丙氨酸转氨酶 c/(μmol·L^-1) ALT c/(μmol·L^-1)	总胆红素 c/(mmol·L^-1) TBIL c/(mmol·L^-1)	肺系数/% Lung coefficient/%	脑系数/% Brain coefficient/%	心系数/% Heart coefficient/%
空白对照组 Control group	36.00±3.43	91.60±12.89	38.20±4.98	0.04±0.74	0.52±0.08	0.60±0.05	0.32±0.02
模型对照组 Model group	39.71±3.59	121.43±25.2	39.86±9.48	0.34±0.31	0.56±0.05	0.66±0.03^aa	0.33±0.03
G₁敏感组 G₁ sensitive group	48.38±8.53^aabb	166.25±87.16^aa	40.87±7.22	0.39±0.28	0.62±0.05^aab	0.71±0.05^aab	0.34±0.02
G₁耐受组 G₁ tolerant group	44.75±6.50^aa	132.75±33.94	39.75±11.85	0.26±0.26	0.55±0.04^c	0.64±0.04^acc	0.33±0.03

Figure 1 HE staining results of heart, lung, and brain tissue sections in rats (×40)Note： The scale bar in this figure is 0.1 mm. Black arrows in the sections of lung represent significant thickening in alveolar walls or infiltration with inflammatory cells. Black arrows in the sections of brain represent a slight disorder in the arrangement of hippocampal cells or cellular vacuolization.

Figure 2 Expression levels of HIF-1 α gene （A） and VEGFA gene （B） in the brain of rats detected by real-time fluorescent quantitative PCRNote：HIF-1α, hypoxia-inducible factor-1α; VEGFA, vascular endothelial growth factor A. Compared with control group, ****P<0.000 1.

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Screening of Hypoxia-Sensitive and Hypoxia-Tolerant Wistar Rats and Preliminary Exploration of Hypoxia Sensitivity in Their G₁Generation

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