Changes of platelet rheological behavior and the interventional effects of ulinastatin in rats with high-voltage electrical burns
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摘要: 目的 探讨高压电烧伤对大鼠血小板聚集数、β血小板球蛋白(β-TG)及血小板因子4(PF-4)的影响以及乌司他丁的干预效果。 方法 将240只SD大鼠按随机数字表法分为假伤组、单纯电伤组、生理盐水组和乌司他丁组,每组60只。以左前肢近心端外侧为电流入口,右后肢近心端外侧为电流出口,单纯电伤组、生理盐水组和乌司他丁组大鼠用调压器及实验变压器造成入口和出口处各1 cm×1 cm的高压电烧伤创面;假伤组大鼠接相同装置但不通电致假伤。伤后2 min,生理盐水组大鼠按2 mL/kg腹腔注射生理盐水,乌司他丁组大鼠按2×104 U/kg腹腔注射10 g/L乌司他丁。伤前15 min及伤后5 min、1 h、2 h、4 h、8 h,每组各取10只大鼠,经心脏采血5~7 mL,取0.05 mL制成活血涂片,观察血小板聚集数;其余血样分离血清,用ELISA法检测血清β-TG和PF-4含量。对数据行析因设计方差分析、SNK检验、Kruskal-Wallis
H 检验、Wilcoxon秩和检验,并进行Bonferroni校正。 结果 (1)伤前15 min,假伤组、单纯电伤组、生理盐水组、乌司他丁组大鼠血小板聚集数相近,分别为(5.9±1.2)、(5.8±1.2)、(5.9±1.3)、(5.9±1.1)个(P 值均大于0.05)。伤后5 min、1 h、2 h、4 h、8 h,单纯电伤组大鼠血小板聚集数分别为(57.2±16.3)、(59.1±16.9)、(60.8±20.6)、(83.6±24.9)、(83.4±30.3)个,较假伤组的(6.0±1.3)、(6.0±1.4)、(5.9±1.4)、(5.7±1.1)、(5.8±1.3)个明显增多(P 值均小于0.001);乌司他丁组大鼠血小板聚集数分别为(29.6±7.4)、(31.9±10.1)、(35.0±14.2)、(43.0±13.6)、(35.2±11.1)个,明显少于生理盐水组的(58.3±16.1)、(63.9±18.0)、(60.8±17.7)、(74.2±23.0)、(82.3±21.9)个(P 值均小于0.001)。假伤组大鼠组内各时相点血小板聚集数比较,差异均无统计学意义(P 值均大于0.05);其余3组大鼠伤后各时相点血小板聚集数均显著多于组内伤前15 min(P 值均小于0.001)。(2)伤后2、4、8 h,单纯电伤组大鼠血清β-TG含量显著高于假伤组(Z 值为-3.780~-3.477,P 值均小于0.05)。伤后5 min、4 h,乌司他丁组大鼠血清β-TG含量显著低于生理盐水组(Z 值分别为-3.477、-3.780,P 值均小于0.05)。假伤组大鼠组内各时相点血清β-TG含量总体比较,差异无统计学意义(χ 2=0.130,P >0.05)。单纯电伤组大鼠伤后2、4、8 h血清β-TG含量显著高于组内伤前15 min(Z 值为-3.780~-3.553,P 值均小于0.05),生理盐水组大鼠伤后5 min、1 h、4 h血清β-TG含量显著高于组内伤前15 min(Z 值为-3.780~-3.477,P 值均小于0.05),乌司他丁组大鼠伤后1、4 h血清β-TG含量显著高于组内伤前15 min(Z 值分别为-3.250、-3.780,P 值均小于0.05)。(3)伤后2、8 h,单纯电伤组大鼠血清PF-4含量显著高于假伤组(P 值均小于0.05)。伤后2 h,乌司他丁组大鼠血清PF-4含量显著高于生理盐水组(P <0.05);伤后4、8 h,乌司他丁组大鼠血清PF-4含量显著低于生理盐水组(P 值均小于0.05)。假伤组大鼠各时相点血清PF-4含量相近(P 值均大于0.05);伤后2、8 h,单纯电伤组大鼠血清PF-4含量显著高于组内伤前15 min(P 值均小于0.05);伤后1、4、8 h,生理盐水组大鼠血清PF-4含量显著高于组内伤前15 min(P 值均小于0.05);伤后1、2、4、8 h,乌司他丁组大鼠血清PF-4含量与组内伤前15 min比较,差异有统计学意义(P 值均小于0.05)。 结论 高压电烧伤大鼠β-TG和PF-4分泌异常,血小板聚集数增多,导致微循环障碍。乌司他丁通过抑制β-TG和PF-4的分泌,减少血小板聚集数,从而减轻高压电烧伤微循环障碍。Abstract: Objective To explore the influence of high-voltage electrical burns on the number of platelet aggregation, β-thromboglobulin (β-TG) and platelet factor 4 (PF-4) and the interventional effects of ulinastatin in rats with high-voltage electrical burns. Methods A total of 240 Sprague-Dawley rats were divided into sham injury (SI) group, simple electrical burn (SEB) group, normal saline (NS) group, and ulinastatin (UTI) group according to the random number table, with 60 rats in each group. The electrical current was applied to the outside proximal part of left forelimb of rats and exited from the outside proximal part of right hind limb of rats. Rats in groups SEB, NS, and UTI were inflicted with high-voltage electrical burn wounds of 1 cm×1 cm at current entrances and exits, with the voltage regulator and experimental transformer. Rats in group SI were sham injured through connecting the same equipments without electricity. At 2 min post injury, rats in group NS were intraperitoneally injected with 2 mL/kg NS, and rats in group UTI were intraperitoneally injected with 2×104 U/kg UTI of 10 g/L. At 15 min before injury and 5 min, 1 h, 2 h, 4 h, 8 h post injury, 10 rats in each group were selected to collect 5-7 mL blood of heart respectively. Blood of 0.05 mL were collected to make fresh blood smear for observing the number of platelet aggregation, and serum were separated from the remaining blood to determine content of β-TG and PF-4 with enzyme-linked immunosorbent assay. Data were processed with analysis of factorial design of variance, student-Newman-Keuls test, Kruskal-WallisH test, Wilcoxon rank sum test, and Bonferroni correction. Results (1) At 15 min before injury, the numbers of platelet aggregation of rats were close among groups SI, SEB, NS and UTI (5.9±1.2, 5.8±1.2, 5.9±1.3, 5.9±1.1, respectively, withP values above 0.05). At 5 min, 1 h, 2 h, 4 h, 8 h post injury, the numbers of platelet aggregation of rats in group SEB were 57.2±16.3, 59.1±16.9, 60.8±20.6, 83.6±24.9, and 83.4±30.3, respectively, obviously more than those in group SI (6.0±1.3, 6.0±1.4, 5.9±1.4, 5.7±1.1, and 5.8±1.3, respectively, withP values below 0.001); the numbers of platelet aggregation of rats in group UTI were 29.6±7.4, 31.9±10.1, 35.0±14.2, 43.0±13.6, and 35.2±11.1, respectively, obviously more than those in group NS (58.3±16.1, 63.9±18.0, 60.8±17.7, 74.2±23.0, and 82.3±21.9, respectively, withP values below 0.001). There was no significantly statistical difference in the number of platelet aggregation of rats in group SI between each two time points within the same group (withP values above 0.05), but the number of platelet aggregation of rats in the other 3 groups at each time point post injury was significantly more than that of the same group at 15 min before injury (withP values below 0.001). (2) At 2, 4, and 8 h post injury, β-TG content of serum of rats in group SEB was significantly higher than that in group SI (withZ values from -3.780 to -3.477,P values below 0.05). At 5 min and 4 h post injury, β-TG content of serum of rats in group UTI was significantly lower than that in group NS (withZ values respectively -3.477 and -3.780,P values below 0.05). There was no significantly statistical difference in β-TG content of serum of rats in group SI at all time points of the same group (χ 2=0.130,P >0.05). At 2, 4, and 8 h post injury, β-TG content of serum of rats in group SEB was significantly higher than that of the same group at 15 min before injury (withZ values from -3.780 to -3.553,P values below 0.05). At 5 min, 1 h, and 4 h post injury, β-TG content of serum of rats in group NS was significantly higher than that of the same group at 15 min before injury (withZ values from -3.780 to -3.477,P values below 0.05). At 1 and 4 h post injury, β-TG content of serum of rats in group UTI was significantly higher than that of the same group at 15 min before injury (withZ values respectively -3.250 and -3.780,P values below 0.05). (3) At 2 and 8 h post injury, PF-4 content of serum of rats in group SEB was significantly higher than that in group SI (withP values below 0.05). At 2 h post injury, PF-4 content of serum of rats in group UTI was significantly higher than that in group NS (P <0.05), and at 4 and 8 h post injury, PF-4 content of serum of rats in group UTI was significantly lower than that in group NS (withP values below 0.05). At all time points, PF-4 content of serum of rats in group SI was close (withP values above 0.05). At 2 and 8 h post injury, PF-4 content of serum of rats in group SEB was significantly higher than that of the same group at 15 min before injury (withP values below 0.05). At 1, 4, and 8 h post injury, PF-4 content of serum of rats in group NS was significantly higher than that of the same group at 15 min before injury (withP values below 0.05). There were significantly statistical differences in PF-4 content of serum of rats between all time points except for 5 min post injury and 15 min before injury (withP values below 0.05). Conclusions Increasing number of platelet aggregation and abnormal secretion of β-TG and PF-4 of rats with high-voltage electrical burns can lead to microcirculation disturbance. UTI can alleviate microcirculation disturbance caused by high-voltage electrical burns by reducing the number of platelet aggregation and inhibiting secretion of β-TG and PF-4.-
Key words:
- Burns, electric /
- Microcirculation /
- Platelet aggregation /
- Platelet factor 4 /
- β-thromboglobulin /
- Ulinastatin
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