中药红景天和血必净对心肺复苏后大鼠脑线粒体功能的影响

Effects of Herba rhodiolae and Xuebijing injection on brain mitochondrial function in rat models after cardiopulmonary resuscitation

目的 探讨红景天和血必净对心搏骤停（CA）大鼠自主循环恢复（ROSC）后脑细胞线粒体功能的影响.方法 选择40只SD大鼠按随机数字表法分为假手术组、肾上腺素组、血必净组、红景天组,每组10只.用改良窒息法复制大鼠CA心肺复苏（CPR）模型,复苏早期肾上腺素组静脉注射肾上腺素0.01 mg/kg,血必净组静脉泵入血必净注射液4 mL/kg,红景天组静脉泵入红景天注射液4 mL/kg.记录基础状态（动静脉置管、气管切开稳定后10 min）、CA时、ROSC时及ROSC后1、2、3、4、5、6h有创平均动脉压（MAP）.ROSC后6h处死小鼠,分离提取各组海马脑组织线粒体,检测各组线粒体呼吸功能,线粒体内、外膜完整性,电镜下观察各组脑组织线粒体超微结构,各组脑组织中丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性.结果 肾上腺素组、红景天组、血必净组CA时MAP均明显低于基础状态时[mmHg（1 mmHg=0.133 kPa）：38±10比122±6,43±5比126±11,44±6比127±9],ROSC时明显高于CA时（63±11比38±10、75±6比43±5、71±9比44±6）,各组其余各时间点MAP水平比较差异均无统计学意义（均P＞0.05）.与假手术组比较,肾上腺素组、红景天组、血必净组在ROSC后6h脑组织线粒体R3、R4、内膜电位、外膜完整率、SOD活性均明显降低[R3 （nmol·L^-1·min^-1· mg^-1）：28.49±2.96、46.15±3.49、44.87±2.18比64.20±4.95;R4（nmol·L^-1·min^-1· mg^-1）：12.54±1.37、11.90±1.06、11.38±0.97比13.80±1.13,线粒体呼吸控制率（RCR）：2.30±0.39、3.91±0.51、3.77±0.81比4.68±0.54,内膜电位（mV）：1.40±0.07、1.74±0.08、1.72±0.07比2.02±0.08,外膜完整率：（31.38±4.03）％、（45.38±4.44）％、（45.40±3.87）％比（66.13±5.11）％;SOD活性（kU/L）：28.54±0.57、32.08±0.66、30.88±0.57比34.91±0.83,均P＜0.05],MDA的含量升高（mmol/L：10.14±0.18、7.45±0.49、6.87±0.98比2.57±0.19,P＜0.05）,但与肾上腺素组比较,血必净组、红景天组脑组织线粒体R3、R4、RCR、内膜电位、外膜完整率、SOD活性明显升高,MDA含量明显降低（均P＜0.05）.电镜下可见假手术组神经细胞超微结构基本正常,染色质分布均匀,胞质内有丰富的粗面内质网和高尔基体,线粒体成椭圆形,嵴排列规整;肾上腺素组、血必净组、红景天组线粒体结构均有不同程度的损伤,尤其是肾上腺素组损伤最重,血必净组、红景天组较肾上腺素组线粒体结构损伤有所恢复.结论 CA大鼠ROSC后脑细胞线粒体功能明显下降,应用血必净和红景天注射液可减少线粒体损伤,部分恢复线粒体功能.

Objective To investigete the effects of Herba rhodiolae injection and Xuehijing injection on brain cell mitochondrial function of rats with cardiac arrest （CA） after restoration of spontaneous circulation （ROSC）. Methods Fogy Sprague-Dawley （SD） rats were divided into sham operation group, epinephrine group, Xuebijing group and Herha rhodiolae group by random number table, 10 rats in each group. The animal model of CA cardiopulmonary resuscitation （CPR） was replicated by improved suffocation method. Early in the resuscitation, in the epinephrine group, epinephrine 0.01 mg/kg was intravenously injected, and Xuebijing 4 mL/kg and Herha rhodiolae 4 mlJkg were respectively infused intravenously by a pump to the corresponding groups. The mean arterial pressure （MAP） levels of the rats under trauma at the basic status （10 minutes after arteriovenous puncture and indwelling catheter and tracheotomy being stabilized）, CA point, ROSC point and 1, 2, S, 4, 5, 6 hours after ROSC were recorded. The rats were sacrificed at the end of 6 hours after ROSC, and the hippocampal brain tissue mitochondria were isolated and extracted in various groups. The mitochondrial respiratory function, the integrity of inner and outer membrane of mitochondrion was examined, and the mitochondrial ultra structures of brain tissues were observed under a transmission electron microscope in the four groups. The content of malonaldehyde （MDA） and superoxide dismutase （SOD） activity in brain tissue were detected in the four groups. Results The MAP at CA point was obviously lower than that at the basic status [mmHg （1 mmHg = 0.133 kPa）： 38±10 vs. 122±6, 43±5 vs. 126±11,44±6 vs. 127±9] and the MAP at ROSC point was obviously higher than that at CA point （63±11 vs. 38 ±10, 75±6 vs. 43±5, 71±9 vs. 44±6） in epinephrine group, Herba rhodiolae group and Xuebijing group; the comparisons of MAP levels among other time points in each group had no statistically significant differences （all P 〉 0.05）. Compared with sham operation group, the brain mitochondrial R3, R4, inner membrane potential, the rate of integrity of milochondrial outer membrane and SOD activity in epinephrine g：roup, Xuebijing injection group and Herba rhodiolae gruup were significantly decreased after 6 hours of ROSC JR3 （nmol·L^-1·min^-1·mg^^-1）： 28.49±2.96, 46.15±3.49, 44.87±2.18 vs. 64.20±4.95 ; R4 （nmol·L^-1·min^-1·mg^^-1）： 12.54±1.37, 11.90±1.06, 11.38±0.97 vs. 13.80±1.13, respiratmy control rate （RCR）： 2.30±0.39, 3.91±0.51, 3.77±0.81 vs. 4.68±0.54, inner membrane potential （mV）： 1.40±0.07, 1.74 ±0.08, 1.72±0.07 vs. 2.02±0.08, rate of integrity of outer membrane： （31.38±4.03）%, （45.38±4.44）%, （45.40 ±3.87）% vs. （66.13±5.11）%; SOD activity （kU/I,）： 28.54±0.57, 32.08±0.66, 30.88±0.57 vs. 34.91±0.83, all P 〈 0.05]; the content of MDA was increased （mmol/L： 10.14±0.18, 7.45±0.49, 6.87±0.98 vs. 2.57±0.19, P 〈 0.05）. But the brain tissue mitochondrial R3, R4, RCR, inner membrane potential, outer membrane integrity rate and SOD activity were significantly elevated and the level of MDA was obviously decreased in Xuebijing injection group and Herba rhodiolae group compared with those in epinephrine group （all P 〈 0.05）. Under transmission electron microscope, in the sham operation group, the ultrastructure of nerve cell was basically normal, the chromatin was well-distributed, almndant rough endoplasmic reticulum and Golgi body were present in cytoplasm, the mitochondrion was oval in shape and its cristae arranged in order; there were different degrees of injury in the structure of mitochondrion in epinephrine group, Xuebijing injection group and Herba Rhodiolae group, especially the damage in epinephrine group was the severest; in contrast to the epinephrine group, the damage of mitochondrion was somewhat recovering in Xuebijing injection group and Herba rhodiolae group. Conclusion The brain mitochondrial function is obviously impaired in rats with CA after ROSC, and application of Xuebijing injection or Herba rhodiolae injection can alleviate mitochondrial injury and partly improve its function.