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中华损伤与修复杂志(电子版)

中华损伤与修复杂志(电子版) ›› 2020, Vol. 15 ›› Issue (01) : 18 -25. doi: 10.3877/cma.j.issn.1673-9450.2020.01.004

所属专题： 文献；

论著

电针足三里穴复合延迟静脉液体复苏对60%血容量失血性休克大鼠的作用研究
钟毓贤1, 张慧苹2, 王海滨3, 吴育寿4, 马丽千2, 张文华2, 姚咏明2, 胡森2,†()   
  1. 1. 100048　北京，解放军总医院第六医学中心康复医学科
    2. 100853　北京，解放军总医院医学创新研究部创伤修复与组织再生研究中心
    3. 100037　北京，解放军总医院第四医学中心检验科
    4. 100037　北京，解放军总医院第四医学中心烧伤研究所
  • 收稿日期:2019-12-23 出版日期:2020-02-01
  • 通信作者: 胡森
  • 基金资助:
    国家自然科学基金项目(81774422)

Effect of electroacupuncture at Zusanli point combined with delayed resuscitation on the hemorrhagic shock rats with 60% blood volume

Yuxian Zhong1, Huiping Zhang2, Haibin Wang3, Yushou Wu4, Liqian Ma2, Wenhua Zhang2, Yongming Yao2, Sen Hu2,†()   

  1. 1. Department of Rehabilitation Medicine, Six Medical Center of PLA General Hospital, Beijing 100048, China
    2. Research Center of Trauma Repair and Tissue Regeneration, Medical Innovation Research Department, Chinese PLA General Hospital, Beijing 100853, China
    3. Department of Laboratory, Fourth Medical Center of PLA General Hospital, Beijing 100037, China
    4. Burn Institute, Fourth Medical Center of PLA General Hospital, Beijing 100037, China
  • Received:2019-12-23 Published:2020-02-01
  • Corresponding author: Sen Hu
  • About author:
    Corresponding author: Hu Sen, Email:
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引用本文：

钟毓贤, 张慧苹, 王海滨, 吴育寿, 马丽千, 张文华, 姚咏明, 胡森. 电针足三里穴复合延迟静脉液体复苏对60%血容量失血性休克大鼠的作用研究[J/OL]. 中华损伤与修复杂志(电子版), 2020, 15(01): 18-25.

Yuxian Zhong, Huiping Zhang, Haibin Wang, Yushou Wu, Liqian Ma, Wenhua Zhang, Yongming Yao, Sen Hu. Effect of electroacupuncture at Zusanli point combined with delayed resuscitation on the hemorrhagic shock rats with 60% blood volume[J/OL]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2020, 15(01): 18-25.

目的

研究电针足三里穴复合延迟静脉液体复苏对60%血容量失血性休克(HS)大鼠生存率、血流动力学、血气指标、脏器血流量和脏器功能的影响。

方法

选取144只SPF级成年雄性大鼠，制作60%血容量HS模型。大鼠麻醉后置管，沿腹中线剖开腹腔长约4 cm，用浸润0.9%氯化钠溶液的纱布覆盖。从股静脉注射1%肝素0.9%氯化钠溶液行全身肝素化后，从股动脉开始抽血。首先在10 min内从股动脉抽出全身血容量的40%，然后在170 min内利用抽液泵从股静脉缓慢抽取全身血容量的20%，总失血量为大鼠全身血容量的60%，HS模型制作完成，记为休克即刻。(1)实验一，选用72只大鼠HS模型，按随机数字表法分成休克不补液组(HS组)、休克后电针组(HS＋EA组)、休克延迟补液组(HS＋DFR组)、休克后电针复合延迟补液组(HS＋EA＋DFR组)。HS组：只进行HS模型制作，不进行针刺和补液。HS＋EA组：于HS模型制作完成后30 min针刺双侧足三里穴，不进行补液；HS＋DFR组：在休克后3 h用3倍失血量的乳酸林格氏液进行股静脉输液30 min，不进行针刺；HS＋EA＋DFR组：HS模型完成后30 min针刺双侧足三里穴，休克后3 h进行与HS＋DFR组相同的静脉延迟补液。计算4组大鼠休克即刻、休克后3、12、24 h生存率；监测造模前30 min，休克即刻，休克后3、12 h平均动脉压(MAP)和腹腔各脏器血流量。(2)实验二，选择余下72只大鼠HS模型，分组及处理同实验一，计算休克后3 h各组动脉血气和脏器功能指标。数据比较采用单因素方差分析或Kruskal-Wallis秩和检验、t检验、log-rank检验。

结果

(1)休克即刻，各组生存率均为100.0%，休克后3 h，HS组、HS＋EA组、HS＋DFR组和HS＋EA＋DFR组大鼠的生存率分别为61.1%、77.8%、77.8%和88.9%，4组比较差异无统计学意义(P>0.05)。休克后12 h，HS＋EA组、HS＋DFR组和HS＋EA＋DFR组大鼠的生存率分别为55.6%、55.6%、61.1%，均显著高于HS组(0)，差异均有统计学意义(t＝6.51、6.73、6.84，P值均小于0.05)。休克后24 h，HS＋EA＋DFR组大鼠生存率(50.0%)显著高于HS＋DFR组(16.7%)和HS＋EA组(11.1%)，差异有统计学意义(t＝2.51、2.17，P值均小于0.05)。(2)休克即刻，4组MAP、肝组织血流量(HBF)、肾组织血流量(RBF)和小肠黏膜血流量(IMBF)较造模前30 min均显著降低，差异均有统计学意义(P值均小于0.05)。休克后3 、12 h，HS组、HS＋EA组、HS＋DFR组和HS＋EA＋DFR组MAP分别为(43.32±5.94)、(64.09±9.64)、(52.85±10.12)、(62.04±7.12) mmHg(1 mmHg＝0.133 kPa)和0、(55.52±11.32)、(67.39±12.03)、(94.78±9.54) mmHg，2个时间点HS组与HS＋EA组比较差异均有统计学意义(t＝3.61、37.00，P值均小于0.05)；HS＋EA＋DFR组与HS＋DFR组比较，差异均有统计学意义(t＝2.01、6.54，P值均小于0.05)；休克后3 h，HS＋EA＋DFR组与HS＋EA组比较，差异无统计学意义(t＝1.04，P>0.05)，休克后12 h，2组比较差异有统计学意义(t＝3.68，P<0.05)。休克后3、12 h，HS组、HS＋EA组、HS＋DFR组和HS＋EA＋DFR组HBF分别为(41.31±4.13)、(47.55±3.21)、(42.54±4.19)、(49.86±4.68) U和0、(52.14±5.53)、(66.24±4.04)、(79.41±7.51) U，2个时间点HS组与HS＋EA组比较差异均有统计学意义(t＝4.16、45.00，P值均小于0.05)；HS＋EA＋DFR组与HS＋DFR组比较，差异均有统计学意义(t＝3.41、3.12，P值均小于0.05)；休克后3 h，HS＋EA＋DFR组与HS＋EA组比较，差异无统计学意义(t＝1.58，P>0.05)，休克后12 h，2组比较差异有统计学意义(t＝3.98，P<0.05)。休克后3、12 h，HS组、HS＋EA组、HS＋DFR组和HS＋EA＋DFR组RBF分别为(81.29±8.49)、(106.48±9.74)、(77.59±8.32)、(100.18±10.48) U和0、(86.81±4.58)、(113.38±10.03)、(158.01±11.63) U, 2个时间点HS组与HS＋EA组比较差异均有统计学意义(t＝3.21、24.00，P值均小于0.05)；HS＋EA＋DFR组与HS＋DFR组比较，差异均有统计学意义(t＝2.67、3.49，P值均小于0.05)；休克后3 h，HS＋EA＋DFR组与HS＋EA组比较，差异无统计学意义(t＝1.55，P>0.05)，休克后12 h，2组比较差异有统计学意义(t＝3.71，P<0.05)。休克后3、12 h，HS组、HS＋EA组、HS＋DFR组和HS＋EA＋DFR组IMBF分别为(43.98±4.75)、(89.92±4.72)、(51.03±6.90)、(94.50±7.61) U和0、(76.65±11.32)、(104.42±12.03)、(143.26±9.54) U，2个时间点HS组与HS＋EA组比较差异均有统计学意义(t＝3.71、30.00，P值均小于0.05)；HS＋EA＋DFR组与HS＋DFR组比较，差异均有统计学意义(t＝2.37、4.38，P值均小于0.05)；休克后3 h，HS＋EA＋DFR组与HS＋EA组比较，差异无统计学意义(t＝1.08，P>0.05)，休克后12 h，2组比较差异有统计学意义(t＝4.74，P<0.05)。(3)休克后3 h，HS组pH、乳酸、动脉血二氧化碳分压、丙氨酸转氨酶、肌酐、二胺氧化酶分别为7.04±0.07、(9.11±1.28) mmol/L、(50.08±3.07) mmHg、(153.15±16.56) U/L、(82.70±7.26) mmol/L、(19.06±2.50) U/L，与HA＋EA组[7.19±0.03、(7.16±1.18) mmol/L、(42.53±4.40) mmHg、(98.26±11.45) U/L、(74.4±6.56) mmol/L、(29.35±2.06) U/L]比较，差异均有统计学意义(t＝8.36、4.75、5.97、11.57、3.60、13.48，P值均小于0.05)；休克后3 h，HS＋DFR组各指标分别为7.04±0.04、(9.06±1.15) mmol/L、(48.14±3.10) mmHg、(136.46±14.24) U/L、(86.5±7.38) mmol/L、(20.56±2.64) U/L，与HS＋EA＋DFR组[7.17±0.14、(7.22±1.07) mmol/L、(40.52±3.09) mmHg、(99.01±10.14) U/L、(72.5±6.41) mmol/L、(25.74±3.20) U/L]比较，差异均有统计学意义(t＝3.79、4.97、7.39、9.09、6.08、5.30，P值均小于0.05)；HS＋EA组与HS＋EA＋DFR组各指标比较，差异均无统计学意义(t＝0.31、0.28、0.33、0.36、0.29、0.35，P值均大于0.05)。

结论

电针足三里穴能显著改善致死性HS模型大鼠的组织灌流并保护脏器功能，提高延迟补液大鼠24 h生存率。

关键词: 休克, 电针, 大鼠, 血气分析, 延迟补液
Objective

To study the effect of electroacupuncture at Zusanli point combined with delayed venous fluid resuscitation on survival rate, hemodynamics, blood gas index, blood flow and organ function of 60% blood volume hemorrhagic shock (HS) rats.

Methods

A total of 144 adult male SPF rats were selected to make a 60% HS model. After anesthesia, the rats were placed into tube, and the abdominal cavity was cut open about 4 cm along the midline of the abdomen, and covered with gauze impregnated with 0.9% sodium chloride solution. After injected 1% heparin saline from the femoral vein for systemic heparin, blood was drawn from the femoral artery. Firstly, 40% of the whole body blood was drawn from the femoral artery within 10 minutes, and then 20% of the whole body blood was slowly drawn from the femoral vein using a suction pump within 170 minutes. The total blood loss was 60% of the whole body blood of the rat. HS model was completed and recorded as shock immediately. (1) In experiment one, seventy-two rat HS models were selected and divided into shock non-hydration group (HS group), shock electroacupuncture group (HS+ EA group), shock delayed fluid replacement group (HS+ DFR group) and electroacupuncture combined delayed fluid replacement group (HS+ EA+ DFR group) according to the random number table method. HS group: only HS model was made, acupuncture and rehydration were not performed. HS+ EA group: acupuncture both sides of Zusanli 30 minutes after the completion of HS model, without rehydration; HS+ DFR group: 3 h after shock, 3 times blood loss of lactated Ringer′s solution for femoral vein infusion for 30 minutes without acupuncture; HS+ EA+ DFR group: 30 minutes after the completion of the HS model, acupuncture both sides of Zusanli point, and 3 hours after shock, the same intravenous delayed rehydration as in the HS+ DFR group was performed. Calculate the immediate, 3, 12, and 24 h aftere shock survival rates of the 4 groups of rats; monitor the mean arterial pressure (MAP) and blood flow in the abdominal organs 30 minutes before the shock, 3, 12 h after shock. (2) Experiment two: 72 rat HS models were selected, grouped and treated in the same way as experiment one, and arterial blood gas and organ function indexes of each group were calculated at 3 h after shock. Data were processed with one-way analysis of variance or Kruskal-Wallis rank sum test, t test, log-rank test.

Results

(1) Immediate shock, the survival rate of each group was 100.0%, and 3 h after shock, the survival rates of rats in the HS group, HS+ EA group, HS+ DFR group, and HS+ EA+ DFR group were 61.1%, 77.8%, 77.8% and 88.9%, there was no statistically significant difference between the 4 groups (P>0.05). At 12 h after shock, the survival rates of the rats in the HS+ EA group, HS+ DFR group, and HS+ EA+ DFR group were 55.6%, 55.6%, and 61.1%, which were significantly higher than that (0) of the HS group, the differences were statistically significant (t= 6.51, 6.73, 6.84; with P values below 0.05). At 24 h after shock, the survival rate of the rats in the HS+ EA+ DFR group was 50.0%, significantly higher than those in the HS+ DFR group (16.7%) and the HS+ EA group(11.1%) (t= 2.51, 2.17; with P values below 0.05). (2) Immediately after shock, the MAP, liver tissue blood flow (HBF), renal tissue blood flow (RBF), and small intestinal mucosal blood flow (IMBF) in the 4 groups were significantly reduced compared with 30 minutes before modeling, and the differences were statistically significant (with P values below 0.05). At 3 and 12 h after shock, the MAP of the HS group, HS+ EA group, HS+ DFR group, and HS+ EA+ DFR group were (43.32±5.94), (64.09±9.64), (52.85±10.12), (62.04±7.12) mmHg (1 mmHg=0.133 kPa) and 0, (55.52±11.32), (67.39±12.03), (94.78±9.54) mmHg, the differences between the HS group and the HS+ EA group at the two time points were statistically significant (t= 3.61, 37.00; with P values below 0.05); the differences between the HS+ EA+ DFR group and the HS+ DFR group were statistically significant (t= 2.01, 6.54; with P values below 0.05); at 3 h after shock, there was no statistically significant difference between the HS+ EA+ DFR group and the HS+ EA group (t=1.04, P>0.05), at 12 h after shock, the difference between the 2 groups was statistically significant (t=3.68, P<0.05). At 3 and 12 h after shock, the HBF in the HS group, HS + EA group, HS + DFR group, and HS+ EA+ DFR group were (41.31±4.13), (47.55±3.21), (42.54±4.19), (49.86±4.68) U and 0, (52.14±5.53), (66.24±4.04), (79.41±7.51) U, there were statistically significant differences between HS group and HS+ EA group at two time points (t=4.16, 45.00; with P values below 0.05); the differences between the HS+ EA+ DFR group and the HS+ DFR group were statistically significant (t=3.41, 3.12; with P values below 0.05); at 3 h after shock, there was no statistically significant difference between HS+ EA+ DFR group and HS+ EA group (t=1.58, P>0.05), at 12 h after shock, the difference between the two groups was statistically significant (t=3.98, P<0.05). At 3, 12 h after shock, the RBF in the HS, HS+ EA, HS+ DFR and HS+ EA+ DFR groups were (81.29±8.49), (106.48±9.74), (77.59±8.32), (100.18±10.48) U and 0, (86.81±4.58), (113.38±10.03), (158.01±11.63) U, there were statistically significant differences between HS group and HS+ EA group at two time points (t= 3.21, 24.00; with P values below 0.05); the difference between the HS+ EA+ DFR group and the HS+ DFR group was statistically significant (t=2.67, 3.49; with P values below 0.05); at 3 h after shock, there was no statistically significant difference between the HS+ EA+ DFR group and HS+ EA group (t=1.55, P> 0.05), and at 12 h after shock, the difference between the two groups was statistically significant (t=3.71, P<0.05). At 3, 12 h after shock, the IMBF of HS group, HS+ EA group, HS+ DFR group and HS+ EA+ DFR group were (43.98±4.75), (89.92±4.72), (51.03±6.90), (94.50±7.61) U and 0, (76.65±11.32), (104.42±12.03), (143.26±9.54) U, the differences between the HS group and the HS+ EA group at two time points were statistically significant (t=3.71, 30.00; with P values below 0.05); the differences between the HS+ EA+ DFR group and the HS+ DFR group were statistically significant (t=2.37, 4.38; with P values below 0.05); at 3 h after shock, there was no statistically significant difference between the HS+ EA+ DFR group and the HS+ EA group (t=1.08, P>0.05), and at 12 h after shock, the difference between the two groups was statistically significant (t=4.74, P<0.05). (3) At 3 h after shock, the pH, lactic acid, partial pressure of arterial carbon dioxide, alanine aminotransferase, creatinine, and diamine oxidase in the HS group were 7.04±0.07, (9.11±1.28) mmol/L, (50.08±3.07) mmHg, (153.15±16.56) U/L, (82.70±7.26) mmol/L, (19.06 ± 2.50) U/L, and HA+ EA group [7.19±0.03, (7.16±1.18) mmol/L, (42.53±4.40) mmHg, (98.26±11.45) U/L, (74.4±6.56) mmol/L, (29.35±2.06) U/L], by comparison, the differences were statistically significant (t=8.36, 4.75, 5.97, 11.57, 3.60, 13.48; with P values below 0.05); at 3 h after shock, each index of the HS+ DFR group was 7.04±0.04, (9.06±1.15) mmol/L, (48.14±3.10) mmHg, (136.46±14.24) U/L, (86.5±7.38) mmol/L, (20.56±2.64) U/L, and HS+ EA+ DFR group [7.17±0.14, (7.22±1.07) mmol/L, (40.52±3.09) mmHg, (99.01±10.14) U/L, (72.5±6.41) mmol/L, (25.74±3.20) U/L], the differences were statistically significant (t=3.79, 4.97, 7.39, 9.09 , 6.08, 5.30; with P values below 0.05); there were no statistically significant differences between HS+ EA group and HS+ EA+ DFR group (t= 0.31, 0.28, 0.33, 0.36, 0.29, 0.35; with P values above 0.05).

Conclusion

Electroacupuncture at Zusanli point can significantly improve tissue perfusion of fatal hemorrhagic shock model and protect organ function, and improve the 24-hour survival rate of rats with delayed fluid replacement.

Key words: Shock, Electroacupuncture, Rats, Blood gas analysis, Delayed rehydration
图1 4组60%血容量HS大鼠Kaplan-Meier生存曲线图。HS　组为休克不补液组；HS＋EA组为休克后电针组；HS＋DFR组为休克后延迟补液组，HS＋EA＋DFR组为休克后电针复合延迟补液组；HS为失血性休克
表1 60%血容量HS前后不同时间点4组大鼠MAP及各脏器血流量的变化(±s)
组别 鼠数(只) MAP(mmHg) HBF(U)
造模前30 min 休克即刻 休克后3 h 休克后12 h 造模前30 min 休克即刻 休克后3 h 休克后12 h
HS组 18 108.03±4.17 29.31±2.31 43.32±5.94 0 91.31±6.32 37.93±6.07 41.31±4.13 0
HS＋EA组 18 109.01±3.87 28.72±2.40 64.09±9.64 55.52±11.32 89.93±8.41 38.15±4.48 47.55±3.21 52.14±5.53
HS＋DFR组 18 107.51±4.37 30.37±2.32 52.85±10.12 67.39±12.03 89.09±7.21 41.51±5.88 42.54±4.19 66.24±4.04
HS＋EA＋DFR组 18 109.15±3.92 28.64±2.57 62.04±7.12 94.78±9.54 92.24±5.10 43.45±6.43 49.86±4.68 79.41±7.51
组别 鼠数(只) RBF(U) IMBF(U)
造模前30 min 休克即刻 休克后3 h 休克后12 h 造模前30 min 休克即刻 休克后3 h 休克后12 h
HS组 18 193.14±16.62 84.44±7.90 81.29±8.49 0 229.36±27.55 66.29±11.07 43.98±4.75 0
HS＋EA组 18 206.42±19.61 82.96±10.21 106.48±9.74 86.81±4.58 222.93±30.32 70.88±8.54 89.92±4.72 76.65±11.32
HS＋DFR组 18 189.24±14.00 85.76±6.44 77.59±8.32 113.38±10.03 231.78±29.90 62.53±7.09 51.03±6.90 104.42±12.03
HS＋EA＋DFR组 18 196.23±16.77 86.03±9.12 100.18±10.48 158.01±11.63 231.41±28.01 65.56±9.32 94.50±7.61 143.26±9.54
表2 休克后3 h各组大鼠血气分析及脏器功能指标比较(±s)
组别 鼠数(只) pH 乳酸(mmol/L) 动脉血二氧化碳分压(mmHg) 动脉血氧分压(mmHg) 丙氨酸转氨酶(U/L) 肌酐(mmol/L) 二胺氧化酶(U/L)
HS组 18 7.04±0.07 9.11±1.28 50.08±3.07 65.53±4.38 153.15±16.56 82.70±7.26 19.06±2.50
HS＋EA组 18 7.19±0.03 7.16±1.18 42.53±4.40 70.55±4.16 98.26±11.45 74.40±6.56 29.35±2.06
HS＋DFR组 18 7.04±0.04 9.06±1.15 48.14±3.10 66.98±4.19 136.46±14.24 86.50±7.38 20.56±2.64
HS＋EA＋DFR组 18 7.17±0.14 7.22±1.07 40.52±3.09 70.14±4.67 99.01±10.14 72.50±6.41 25.74±3.20
t1值 ? 8.36 4.75 5.97 3.53 11.57 3.60 13.48
P1值 ? <0.05 <0.05 <0.05 <0.05 <0.05 <0.05 <0.05
t2值 ? 3.79 4.97 7.39 2.14 9.09 6.08 5.30
P2值 ? <0.05 <0.05 <0.05 0.04 <0.05 <0.05 <0.05
t3值 ? 0.31 0.28 0.33 0.43 0.36 0.29 0.35
P3值 ? >0.05 >0.05 >0.05 >0.05 >0.05 >0.05 >0.05
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