急性减压缺氧对大鼠血液学指标的影响.pdf

第 40 卷第 5 期2023 年 10 月实验动物科学LABORATORY ANIMAL SCIENCEVol.40No.5October 2023论著收稿日期:2023-03-06基金项目:后勤科研计划面上项目(CWH17J010)作者简介:孟盼盼(1995),女,硕士研究生,研究方向:高原缺氧药物研究.E-mail:18693115075 通信作者:赵秀华(1972),女,副主任技师,研究方向:主要从事高原疾病防控和实验工作.E-mail:zhaoxiuhua2021xz 急性减压缺氧对大鼠血液学指标的影响孟盼盼1 谭宏强1 石志群1 张朋朋1 张 洁1 赵秀华2(1.中国人民解放军联勤保障部队第 940 医院药剂科,兰州730050)(2.中国人民解放军西部战区疾病预防控制中心传染病防控二科,成都610000)摘要:目的探讨模拟急性高原低压性缺氧对大鼠血液指标变化的影响。方法建立高原缺氧大鼠损伤模型,将40 只 Wistar 雄性大鼠随机分为常氧对照组、缺氧 10 h 组、缺氧 20 h 组、缺氧 30 h 组,每组 10 只,对照组置于当地海拔 1 500 m 处,缺氧 10、20、30 h 组放入大型低压低氧动物实验舱,模拟海拔 8 000 m 高原低压性缺氧环境,检测大鼠血气、血常规和血生化指标,观察不同缺氧时间对大鼠血液学指标的影响。结果随着缺氧时间的延长,与常氧对照组比较,血气相关指标二氧化碳分压(PaCO2)、氧分压(PaO2)、肺动脉血氧饱和度(SaO2)逐渐下降;血常规指标白细胞(WBC)、红细胞计数(RBC)和血小板(PLT)均显著增加;血生化指标血糖水平显著升高,白蛋白(ALB)、血清尿素氮(BUN)异常升高,肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)显著降低(P0.01 或 P0.05)。结论模拟高原低压性缺氧不同时间大鼠的血液学指标均发生显著改变,严重影响大鼠的正常生理活动。关键词:高原缺氧;血常规;血气;血生化;氧化应激中图分类号:Q95-3文献标志码:A文章编号:1006-6179(2023)05-0048-05DOI:10.3969/j.issn.1006-6179.2023.05.009Effects of Hematological Parameters by Acute Decompression Hypoxia in RatsMENG Panpan1,TAN Hongqiang1,SHI Zhiqun1,ZHANG Pengpeng1,ZHANG Jie1,ZHAO Xiuhua2(1.Key Laboratory of the Plateau Medicine,the 940th Hospital of Joint Logistics Support Force of Chinese Peoples Liberation Army,Lanzhou 730050,China)(2.the Second Department of Infection Disease Control and Prevention,Center for Disease Control andPrevention,Western War Zone,Chinese Peoples Liberation Army,Chengdu 610000,China)Abstract:ObjectiveTo investigate the effects of simulated plateau acute decompression hypoxia on blood indexes in rats.MethodForty healthy male Wistar rats were randomly divided into normoxia control group,hypoxia 10,20,30 h group,10 rats in each group.were put into hypobaric hypoxia treatment to simulate the low-pressure and hypoxic environment at 8 000 m altitude.Normal rats were kept at 1 500 m above sea level,the other 3 groups stayed at simulated altitude of 8 000 m for 10,20 and 30 h respectively by large low pressure and low oxygen animal laboratory.Check blood sugar,blood gas,blood routine,and blood biochemistry.Observe the effects of different hypoxia time on hematological indexes of rats.ResultWith the prolongation of hypoxia time,the blood glucose of the rats also increased;According to the blood gas index,the partial pressure of carbon dioxide(PaCO2),partial pressure of oxygen(PaO2),pulmonary oxygen saturation(SaO2)gradually decrease with the prolongation of hypoxia;Blood routine result showed that white blood cells(WBC),red blood cell count(RBC)and platelet(PLT)in rat arterial blood increased significantly with the prolongation of hypoxia;第 5 期孟盼盼等:急性减压缺氧对大鼠血液学指标的影响 blood biochemical result showed that albumin(ALB),serum urea nitrogen(BUN)were abnormally elevated.creatine kinase(CK),and creatine kinase isoenzyme(CK-MB)were significantly decreased(P 0.01 or P 0.05);缺氧 10、20、30 h 组PCO2、PO2、SO2均低于常氧对照组,且随着缺氧时间的延长显著性降低(P0.05,P0.01);缺氧 10、20、30 h 组 HCO3-、TCO2均低于常氧对照组,且随着缺氧时间的延长呈先增加后降低的趋势(P0.01);缺氧 10、20、30 h 组 cNa+与常氧对照组相比呈上升趋势,且随着缺氧时间的延长逐渐升高(P0.05,P0.01)。缺氧 10 h 组 cK+与常氧对照组相比呈下降趋势,且具有统计学差异(P0.01),随着时间的延长,缺氧 20、30 h 后 cK+上升,与常氧对照组相比没有显著性差异,见表 1。表 1急性减压缺氧大鼠血气分析各项指标对比(xs,n=10)Table 1Comparison of various indexes of blood gas analysis with acute decompression hypoxia in rats(xs,n=10)组别pHHCO3-(mmol/L)TCO2(mmHg)cNa+(mmol/L)常氧对照组7.430.0422.571.2023.601.19142.001.48缺氧 10 h 组7.410.0216.800.69#17.600.71#142.871.85缺氧 20 h 组7.430.0316.971.15#17.761.16#144.700.85#缺氧 30 h 组7.420.0312.101.69#12.841.73#145.081.20#组别cK+(mmol/L)PCO2(mmHg)PO2(mmHg)SO2(%)常氧对照组4.070.2833.391.4998.314.9897.700.37缺氧 10 h 组3.760.15#28.855.96#76.158.42#95.140.66#缺氧 20 h 组4.110.1925.560.98#74.313.40#94.192.27#缺氧 30 h 组4.150.2823.352.68#64.964.58#91.041.61#注:HCO3-为碳酸氢根,TCO2为二氧化碳总量,cK+为 K+浓度,cNa+为 Na+浓度,SO2为血氧饱和度,PO2为氧分压,PCO2 为二氧化碳分压;与常氧对照组相比,#P0.05,#P0.01。Note:HCO3-is bicarbonate,TCO2 is total carbon dioxide,cK+is K+concentration,cNa+is Na+concentration,SO2 is blood oxygen saturation,PO2 is partial pressure of oxygen,PCO2 is partial pressure of carbon dioxide;Compared with the normoxic control group,#P0.05,#P0.01.2.2急性减压缺氧不同时间大鼠血常规指标的影响缺氧 10、20、30 h 组 WBC、RBC、HGB、MCHC、PLT、HCT 与常氧对照组相比呈上升趋势,且随着缺氧时间的延长显著性升高(P0.05,P0.01);缺氧10、20 h 组 MCH 与常氧对照组相比没有显著性差异,至缺氧 30 h 后下降与常氧对照组相比有显著性差异(P0.05)。缺氧 10、20 h 组 MCV 与常氧对照组相比呈显著性下降趋势(P0.01),缺氧 30 h 后上升至与常氧对照组相比无显著性差异,见表 2。表 2急性减压缺氧大鼠血常规各项指标对比(xs,n=10)Table 2Comparison of various indexes of blood routine analysis with acute decompression hypoxia in rats(xs,n=10)组别WBC(109/L)RBC(1012/L)HGB(g/L)HCT(%)常氧对照组4.220.799.410.74172.3012.9448.593.32缺氧 10 h 组2.820.71#10.280.57#189.609.88#52.062.55#缺氧 20 h 组5.820.47#10.440.41#200.7311.04#54.162.62#缺氧 30 h 组10.622.58#10.520.41#207.908.44#54.152.05#组别MCV(fL)MCH(pg)MCHC(g/L)PLT(109/L)常氧对照组51.690.9818.320.18354.404.531544.50197.87缺氧 10 h 组50.310.62#18.310.20364.403.17#1629.80306.73缺氧 20 h 组49.850.94#18.330.24370.454.34#1669.00107.42缺氧 30 h 组52.211.1118.110.23#370.704.49#1709.8873.56#注:WBC 为白细胞,RBC 为红细胞计数,HGB 为血红蛋白,HCT 为红细胞压积,MCV 为平均红细胞体积,MCH 为平均血红蛋白量,MCHC为平均血红蛋白浓度,PLT 为血小板;与常氧对照组相比,#P0.05,#P0.01。Note:WBC is white blood cell,RBC is red blood cell count,HGB is hemoglobin,HCT is hematocrit,MCV is mean red blood cell volume,MCH is mean hemoglobin amount,MCHC is mean hemoglobin concentration,PLT is platelet.Compared with the normoxic control group,#P0.05,#P0.01.2.3急性减压缺氧不同时间对大鼠血生化指标的影响缺氧 10、20、30 h 组 CK、CK-MB 与常氧对照组相比呈显著性下降(P0.05,P0.01);缺氧 10 和20 h 组 LDH 与常氧对照组相比呈显著性上升(P0.01),但缺氧 30 h 后逐渐下调,与常氧对照组相比05第 5 期孟盼盼等:急性减压缺氧对大鼠血液学指标的影响 无显著性差异;缺氧 20、30 h 组 GLU、BUN 与常氧对照组相比呈显著性上升(P 0.05,P 0.01);缺氧10 h 组 ALB 与常氧对照组相比呈下降趋势但无显著性差异,缺氧 20 h 后逐渐上升,至缺氧 30 h 后与常氧对照组相比呈显著上升(P 0.01);缺氧 10、20 h 组 CRE 呈下降趋势,30 h 后升高(P0.05,P0.01);缺氧 30 h 组 UA 与正常对照组相比呈显著上升(P0.01),见表 3。表 3急性减压缺氧大鼠血生化各项指标对比(xs,n=10)Table 3Comparison of various indexes of blood biochemistry analysis withacute decompression hypoxia in rats(xs,n=10)组别CK(U/L)CK-MB(U/L)LDH(mmol/L)GLU(mmol/L)常氧对照组557.0091.111 429.12309.151 586.83338.779.092.00缺氧 10 h 组1 348.13265.18#3 117.09501.32#2 900.00579.89#11.000.54缺氧 20 h 组1 137.71184.89#2 678.60414.05#3 707.90707.22#11.111.02#缺氧 30 h 组891.171194.41#2 349.15446.21#1 896.50225.5913.111.63#组别ALB(g/L)BUN(mmol/L)CRE(mol/L)UA(mol/L)常氧对照组26.932.287.331.7538.601.76102.3315.34缺氧 10 h 组25.890.926.400.7423.911.34#106.317.74缺氧 20 h 组27.722.198.800.47#33.211.77#100.8212.99缺氧 30 h 组30.991.29#11.760.88#41.131.87#126.189.39#注:CK 为肌酸激酶,CK-MB 为肌酸激酶同工酶,LDH 为乳酸脱氢酶,GLU 为血糖,ALB 为白蛋白,BUN 为血清尿素氮,CRE 为肌酐,UA 为尿酸;与常氧对照组相比,#P0.05,#P0.01。Note:CK is creatine kinase,CK-MB is creatine kinase isoenzyme,LDH is lactate dehydrogenase,GLU is blood sugar,ALB is albumin,BUN is serum urea nitrogen,CRE is creatinine,UA is uric acid;Compared with the normoxic control group,#P0.05,#P0.01.3讨 论高原低压性缺氧干扰机体正常的氧化反应,造成细胞代谢和功能紊乱并对机体的组织、器官造成损害。研究报道,缺氧会导致机体葡萄糖稳态严重受损6,血糖异常增加主要是由 细胞功能紊乱引起的胰岛素分泌受损或外周组织中胰岛素抵抗增加7。高血糖是机体微血管和大血管并发症的主要危险因素,会诱发视网膜病变、神经疾病和心血管疾病,威胁着患者的生活质量和寿命8。本实验结果所示,随着缺氧时间的延长,机体血糖逐渐升高,说明机体葡萄糖稳态的受损程度可能与缺氧时间相关。缺氧是影响高原个体健康和生命活动的关键因素,它对生理有重要影响,诱导机体病变9。缺氧与机体药代动力学密切相关,例如:低氧血症和酸性失衡对药 物的吸收、分布、代谢和 排泄有显著 影响10-11。因此,高海拔地区大鼠动脉血气分析对于高原药代动力学具有重要意义。本研究评估了在模拟高原海拔 8 000 m 缺氧 10、20、30 h 后,大鼠的呼吸状态和代谢性酸碱失调12。结果表明,实验大鼠缺氧损伤后,pH 值无明显变化,而 HCO3-、TCO2和PCO2在缺氧 10、20、30 h 后逐渐下降,表明大鼠在缺氧条件下,肺通气过度,存在呼吸性碱中毒,从侧面反映出缺氧导致机体肺呼吸功能受损。此外,电解质 cNa+和 cK+具有调节渗透压和酸碱平衡的作用,二者随着缺氧时间的增长逐渐升高,说明在缺氧条件下,机体酸碱平衡失调13。PO2和 SO2反应了机体血液的带氧率,最大氧气吸收速率的陆地哺乳动物,在稳定状态下的剧烈运动中也很少发生低氧血症14。在缺氧 10、20、30 h 后,PO2和 SO2逐渐下降,客观反映了在高原缺氧环境下,大鼠呼吸功能障碍,组织细胞失去了从血液中摄取氧气的能力,导致机体器官受损。简单的血常规检查及分类检查,可以观察到机体血液红细胞、白细胞、血小板的变化,了解高原缺氧环境下机体的损伤情况,对高原缺氧损伤的初步判定、严重程度以及预测机体缺氧后器官的损伤程度的评估具有重要意义15。血常规结果显示:缺氧损伤后,大鼠为了适应高原环境,随着缺氧时间的延长显著增加 RBC、HGB、HCT、MCV、MCH 和 MCHC来提高机体的携氧能力,维持机体的正常生理功能。但是,RBC 的大量增多导致血液的黏稠度大幅度增加,提高了机体心肌梗死的发病率,同时,WBC 和PLT 与正常对照组相比显著增加,提示了缺氧可能会导致大鼠心脑血管的病变16。生化指标可以评估机体的心脏、肝、肾功能,直15 实验动物科学40 卷观准确的评价在缺氧情况下机体器官的损伤情况。ALB 为反映肝功能的重要指标,缺氧后 ALB 显著升高表明在高原缺氧环境中,大鼠红细胞增多导致血液黏滞,从而影响肝发生病变。缺氧刺激肾产生红细胞生成素,使体内红细胞增多,血液黏稠度增高,外围血管阻力加大,心脏负担加重,引起或加重心力衰竭,诱发心肌梗死、脑血栓等疾病;BUN、CRE、UA为评价肾功能的指标,在缺氧条件下,BUN、CRE、UA 随着缺氧时间的延长显著升高,提示缺氧可能导致肾血管收缩,从而导致肾功能不全17。CK、CK-MB 是反映心功能的重要指标,二者随着缺氧时间延长显著降低,且在急性缺氧 10 h 时损伤最剧烈,提示缺氧导致心肌供氧不足,心肌收缩力下降,出现心功能不全18。但在缺氧 20、30 h 后与缺氧10 h 相比有下降趋势,表明机体对缺氧进行适应性调节,但是仍不可能使之降到正常水平。缺氧后GLU 和 LDH 均升高,说明机体在缺氧环境下血糖增高,无氧呼吸增多,诱导机体氧化应激,随着缺氧时间的增长,LDH 有降低趋势,表明机体对缺氧进行适应性调节,但是仍不可能达到正常水平。综上所述,缺氧会引起机体血液学发生病变,且随着缺氧时间的延长损伤加重,本研究发现血液学变化与心、肝、肾等器官息息相关,因此,本篇文章主要观察了缺氧对机体血液学的变化,以期确定影响这些指标变化的因素和条件,找出合理的改善方法,为后期的实验奠定基础。参 考 文 献 1 LI C,LI X,LIU J,et al.Investigation of the differences between the Tibetan and Han populations in the hemoglobin oxygen affinity of red blood cells and in the adaptation to high-altitude environments J.Hematology,2017,23(5):309.2 WANG R,SUN Y,YIN Q,et al.The effects of metronidazole on cytochrome P450 activity and expression in rats after acute exposure to high altitude of 4300 m J.Biomed,Pharmacother,2017,1(85):296-302.3 PU P,ZHAO 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