基于正交设计探讨复方蕨麻胶囊对低温低氧失血性休克大鼠微循环的影响

Exploring the effects of compound Juema Capsules on microcirculation in hemorrhagic shock rats with low temperature and hypoxia based on orthogonal design

[目的]基于正交设计,探究不同条件(空气氧含量、环境温度及药物浓度)对失血性休克(HS)大鼠微循环的影响,为预防低温低氧HS提供最优方案。[方法]本研究选取海拔(用空气氧含量模拟)[3551.7 m(13.5%)、3080.7 m(14.5%)、2751 m(15.2%)]、环境温度(5~10℃、10~15℃、15~20℃)及药物浓度[复方蕨麻胶囊低(27 mg/kg)、中(54 mg/kg)、高(81 mg/Kg)剂量]三个因素,每个因素各选取三个水平,采用三因素三水平的正交设计,按照L9(34)正交表进行实验,共得出9种组合。将27只350~400 g雄性SD大鼠根据正交设计的干预方案分为9组,每组重复3次,进行HS大鼠的造模。实验过程中采集并记录放血前(T)、复苏前(T0)、复苏后1 h(T1)、复苏后2 h(T2)、复苏后3 h(T3)、复苏后4 h(T4)、复苏后5 h(T5)共7个时间点的平均动脉压(MAP)、心率(HR)、肠系膜微循环指标[包括血管流速、灌注血管密度(PVD)和微循环血流指数(MFI)]。采用相关的统计学方法进行分析,先筛选出实验最佳观测时间点,而后选出本实验条件下对改善HS大鼠的最优方案。[结果]筛选实验最佳观测时间点:对9组大鼠的MAP、HR、肠系膜流速七个时间点进行比较。1)MAP:①各组大鼠的MAP在不同时间点变化趋势一致(F=1.471,P=0.051);②组间差异无统计学意义(F=1.002,P=0.467);③各组MAP在不同时间点不全相等(F=417.804,P<0.001)。2)HR:①各组大鼠的HR在不同时间点变化趋势一致(F=0.782,P=0.730);②组间差异无统计学意义(F=0.647,P=0.729);③各组HR在不同时间点不全相等(F=947.477,P<0.001)。3)肠系膜血管流速:①各组大鼠的肠系膜流速在不同时间点变化趋势不同(F=4.933,P<0.001);②组间流速差异无统计学意义(F=2.141,P=0.086);③各组T在不同时间点不全相等(F=82.425,P<0.001)。4)小结:根据以上数据选定T1为最佳实验观测点。本实验条件下最优方案的选择:①改善血管流速的最佳组合为第二组;②改善PVD的最佳组合为第二组;③改善MFI的最佳组合为第二组;④小结:第二组(即氧含量13.5%、温度10~15℃、药物浓度为中剂量)在实验条件内的组合最佳。[结论]HS大鼠复苏后的五个观察点内,T1这个时间点各指标改善最优;复方蕨麻胶囊中剂量对3500 m左右的HS大鼠复苏效果最佳。

[Objective]Based on orthogonal design,to investigate the effects of different conditions(air oxygen content,ambient temperature and drug concentration)on microcirculation in rats with hemorrhagic shock(HS)and to provide an optimal solution for the prevention of plateau hemorrhagic shock.[Methods]In this study,three factors,altitude(simulated with air oxygen content)[3551.7 m(13.5%),3080.7 m(14.5%),2751 m(15.2%)],ambient temperature(5-10℃,10-15℃,15-20℃)and drug concentration[low(27 mg/kg),medium(54 mg/kg)and high(81 mg/kg)dose of compound Juema Capsule]were selected,and three levels of each factor were chosen,and the experiment was carried out by using a three-factor,three-level orthogonal design in accordance with the orthogonal table of L9(34)to obtain a total of nine Combinations.Twenty-seven 350-400 g male SD rats were divided into nine groups according to the orthogonal design of the intervention scheme,and each group was repeated three times for the modeling of HS rats.Mean arterial pressure(MAP),heart rate(HR),and mesenteric microcirculation indexes[including vascular flow velocity,perfusion vascular density(PVD)and microcirculation flow index(MFI)]were collected and recorded at seven time points during the experiment:before bloodletting(T),before resuscitation(T0),1 h after resuscitation(T1),2 h after resuscitation(T2),3 h after resuscitation(T3),4 h after resuscitation(T4),and 5 h after resuscitation(T5).Relevant statistical methods were used to analyze the results,first to screen the optimal observation time point of the experiment,and then to select the optimal solution for improving the plateau HS rats under the present experimental conditions.[Results]Screening for the best observation time point of the experiment:seven time points of MAP,HR,and mesenteric flow rate were compared among nine groups of rats.1)MAP:①The trend of MAP changes in each group of rats at different time points is consistent(F=1.471,P=0.051);②There was no statistically significant difference between groups(F=1.002,P=0.467);③The MAP values of each group were not all equal at different time points(F=417.804,P<0.001).2)HR:①The HR of each group of rats showed consistent trends at different time points(F=0.782,P=0.730);There was no statistically significant difference between groups(F=0.647,P=0.729);③The HR values of each group were not all equal at different time points(F=947.477,P<0.001).3)Mesenteric flow velocity:①The trend of changes in mesenteric flow velocity among different groups of rats at different time points is different(F=4.933,P<0.001).②There was no statistically significant difference in inter group flow velocity(F=2.141,P=0.086);③At different time points,each group of T was not completely equal(F=82.425,P<0.001).4)Summary:Based on the data,T1 was selected as the best experimental observation point.Selection of the optimal solution for the conditions of this experiment:①The best combination for improving blood vessel flow rate is the second group;②The best combination for improving PVD is the second group.③The best combination to improve MFI is the second group.④Summary:The second group(i.e.oxygen content of 13.5%,temperature of 10-15℃,and drug concentration of medium dose)had the best combination within the experimental conditions.[Conclusion]Among the five observation points after resuscitation of HS rats,T1 is the best observation point in the experiment;the medium dose of compound Juema capsule is the best resuscitation effect for HS rats around 3500 m.