清瘟败毒饮对脓毒症大鼠血清降钙素原及C-反应蛋白水平的影响

An experimental study of influence of Qingwen Baidu decoction on serum procalcitonin and C-reactive protein levels in septic rats

目的：观察清瘟败毒饮对脓毒症大鼠血清降钙素原（PCT）和C-反应蛋白（CRP）水平的影响，探讨清热解毒法治疗脓毒症的机制。方法将50只雄性SD大鼠按随机数字表法分为对照组（n＝5）、模型组（n＝25）及清热解毒组（n＝20）。采用腹腔注射脂多糖（LPS）5 mg/kg制备脓毒症大鼠模型；对照组给予等量生理盐水。清热解毒组于制模后2 h首次给予清瘟败毒饮（生石膏30 g，生地黄10 g，水牛角15 g，黄连4 g，山栀子5 g，桔梗5 g，黄芩5 g，知母5 g，赤芍5 g，玄参5 g，连翘5 g，甘草5 g，牡丹皮5 g，竹叶5 g）0.01 mL/g，其余给药时间为每日08：00-09：00，每日1次；模型组给予等量温开水灌胃。对照组、模型组及清热解毒组于制模后不同时间点各取5只大鼠经腹主动脉取血，采用酶联免疫吸附试验（ELISA）检测大鼠血清PCT、CRP水平；留取肺、肠组织标本，光镜下观察组织病理学改变。结果与对照组比较，制模后2 h模型组PCT水平即明显增高（ng/L：332.32±22.85比70.46±3.18，P＜0.01），并持续到制模后72 h。制模后2、8、48 h对照组和模型组CRP水平比较差异无统计学意义（均P＞0.05），制模后24 h、72 h模型组明显低于对照组（μg/L：281.34±32.81、237.84±41.42比350.09±56.67，P＜0.05和P＜0.01）。制模后8 h、24 h模型组和清热解毒组PCT比较差异无统计学意义（均P＞0.05），清热解毒组制模后48 h起PCT明显低于模型组（ng/L：321.57±28.00比358.12±10.14，P＜0.05），并持续到制模后72 h（ng/L：269.50±49.10比347.69±26.90，P＜0.05）。清热解毒组制模后8 h起CRP水平即明显低于模型组（μg/L：232.73±13.29比335.35±53.78，P＜0.05），制模后72 h两组比较差异仍有统计学意义（μg/L：177.31±6.70比237.84±41.42，P＜0.05）。与模型组比较，清热解毒组肺组织炎性细胞浸润减少，肠黏膜炎症及间质水肿减轻。结论清热解毒法可有效降低LPS诱导的脓毒症大鼠血清PCT、CRP水平；改善炎性细胞在组织中的浸润，从而起到保护组织器官的作用。

Objective To observe the influence of Qingwen Baidu decoction （QBD） on serum procalcitonin （PCT） and C-reactive protein （CRP） levels in septic rats and study the mechanism of heat-clearing and detoxifying method for treatment of sepsis. Methods Fifty male Sprague-Dawley （SD） rats were randomly divided into control group （n=5）, model group （n=25）, and heat-clearing and detoxifying experimental group （experimental group, n=20）. The septic model was reproduced by intra-peritoneal injection of lipopolysaccharide （LPS, 5 mg/kg）. In control group, an equal volume of normal saline was given. After modeling for 2 hours, the heat-clearing and detoxifying experimental group received QBD the first time （composition of the decoction： Gypsum Fibrosum Recens 30 g, Rehmanniae Radix 10 g, Bubali Cornu 15 g, Coptidis Rhizoma 4 g, Gardeniae Fructus 5 g, Platycodonis Radix 5 g, Scutellariae Radix 5 g, Anemarrhenae Rhizoma 5 g, Paeoniae Radix Rubra 5 g, Scrophulariae Radix 5 g, Forsythiae Fructus 5 g, Glycyrrhizae Radix 5 g, Moutan Cortex 5 g, Lophatheri Herba 5 g） by gavage （0.01 mL/g）; the rest administration time was 08：00 to 09：00, once a day. The rats in model group were given an equal volume of warm water by gavage. At different time points after modeling, the blood of 5 rats in control group, model group, and experimental group was collected from the abdominal aorta. The serum PCT and CRP levels were tested by the enzyme linked immunosorbent assay （ELISA）, and the pathological changes in lung and intestinal tissue were observed under a light microscope. Results Compared with the control group, the PCT level of the model group after modeling for 2 hours was significantly increased （ng/L：332.32±22.85 vs. 70.46±3.18, P〈0.01）, this situation continued until 72 hours after modeling. CRP level in the comparison between the control and model groups did not have statistical significant difference at 2, 8, and 48 hours after modeling （all P 〉 0.05）. The level of CRP in model group was lower than that of control group at 24 hours and 72 hours after modeling （μg/L：281.34±32.81, 237.84±41.42 vs. 350.09±56.67, P〈0.05 and P〈0.01）. There were no statistically significant differences in PCT levels between model group and experimental group after modeling for 8 hours and 24 hours （both P 〉 0.05）. The PCT level of experimental group was significantly lower than that of model group beginning from 48 hours after modeling （ng/L： 321.57±28.00 vs. 358.12±10.14, P 〈 0.05）, and this situation continued until 72 hours after modeling （ng/L： 269.50±49.10 vs. 347.69±26.90, P 〈0.05）. The CRP level of experimental group was significantly lower than that of model group beginning from 8 hours after modeling （μg/L：232.73±13.29 vs. 335.35±53.78, P〈0.05）, this statistical significant difference between the two groups persisted until 72 hours after modeling （μg/L：177.31±6.70 vs. 237.84±41.42, P〈0.05）. Compared to those in the model group, the lung tissue inflammatory cell infiltration, the intestinal mucosal inflammation and interstitial edema were milder in the experimental group. Conclusion Heat-clearing and detoxifying therapy can effectively reduce the serum PCT and CRP levels of septic rats induced by LPS, and it can alleviate the infiltration of inflammatory cells in lung tissues so as to play a role in protection of tissue organ.