改良冬眠合剂对严重烫伤大鼠脏器功能的影响

于家傲; 王际壮; 王家强; 章雄; 刘琰

doi:10.3760/cma.j.cn501225-20230329-00105

改良冬眠合剂对严重烫伤大鼠脏器功能的影响

doi: 10.3760/cma.j.cn501225-20230329-00105

上海交通大学医学院附属瑞金医院灼伤整形科，上海　　200025

基金项目:

国家自然科学基金面上项目 82072173, 82172199

上海市生物医药领域定向项目 22DX1900600

上海市临床重点专科项目 shslczdzk02302

上海市综合医院中西医结合专项研究基金 ZHYY-ZXYJHZX-201911

详细信息

通讯作者:
刘琰，Email：rjliuyan@126.com

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出版历程
- 收稿日期: 2023-03-29

Effects of modified lytic cocktail on organ function of severely scalded rats

Department of Burn, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

Funds:

General Program of National Natural Science Foundation of China 82072173, 82172199

Shanghai Directed Projects of Biopharmaceutical Field 22DX1900600

Shanghai Municipal Key Clinical Specialty of China shslczdzk02302

Shanghai Specialized Research Fund for Integrated Chinese and Western Medicine in General Hospitals ZHYY-ZXYJHZX-201911

More Information

Corresponding author: Liu Yan, Email: rjliuyan@126.com

摘要

摘要: 目的对比改良冬眠合剂和传统冬眠合剂对严重烫伤大鼠脏器功能的影响。方法采用实验研究方法。取24只约10周龄雄性SD大鼠，按照随机数字表法分为假伤组、单纯烫伤组、传统冬眠合剂组和改良冬眠合剂组，每组6只。于单纯烫伤组、传统冬眠合剂组和改良冬眠合剂组大鼠背部造成30%体表总面积（TBSA）Ⅲ度烫伤，假伤组大鼠模拟致伤过程致假伤。伤后即刻，传统冬眠合剂组大鼠腹腔注射12 mL/kg由哌替啶、氯丙嗪和异丙嗪构成的传统冬眠合剂，辅以生理盐水灌胃；改良冬眠合剂组大鼠腹腔注射2 mL/kg由咪达唑仑和芬太尼构成的混合药剂，辅以西替利嗪灌胃。随后4组大鼠均腹腔注射乳酸林格液进行补液复苏，使补液及给药总剂量为2 mL·kg ^-1·%TBSA ^-1。次日，对2个冬眠合剂组大鼠再次同前给药。伤后3 d，取各组大鼠腹主动脉血并留取肝脏、小肠和肺脏组织，采用酶联免疫吸附测定法检测血浆中白细胞介素1β（IL-1β）、IL-10和IL-6水平，采用全自动生化分析仪检测血浆中丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、γ-谷氨酰转移酶（γ-GT）、碱性磷酸酶（ALP）、乳酸脱氢酶（LDH）、LDH同工酶1（LDH-1）、肌酸激酶、肌酸激酶同工酶（CK-MB）、尿素、肌酐和尿酸水平，对肝脏、小肠和肺脏组织行苏木精-伊红染色后观察组织学变化。对数据行单因素方差分析和Tukey检验。结果伤后3 d，传统冬眠合剂组大鼠血浆中IL-10水平为（44±16）pg/mL，显著高于改良冬眠合剂组的（20±9）pg/mL和单纯烫伤组的（21±6）pg/mL（ P值均<0.05）；4组大鼠血浆中IL-1β和IL-6水平比较，差异均无统计学意义（ P>0.05）。伤后3 d，单纯烫伤组大鼠血浆中ALT和AST水平分别为（77±14）、（213±65）U/L，均显著高于假伤组的（59±5）、（108±10）U/L（ P<0.05）；改良冬眠合剂组大鼠血浆中ALT和AST水平分别为（61±3）、（116±11）U/L，均显著低于传统冬眠合剂组的（81±13）、（207±54）U/L（ P<0.05），且改良冬眠合剂组大鼠血浆中AST水平显著低于单纯烫伤组（ P<0.05）。伤后3 d，4组大鼠血浆中γ-GT、ALP、LDH、LDH-1、肌酸激酶、CK-MB、肌酐和尿酸水平比较，差异均无统计学意义（ P>0.05）；虽然4组大鼠血浆中尿素水平总体比较差异有统计学意义（ P<0.05），但单纯烫伤组分别与假伤组、传统冬眠合剂组、改良冬眠合剂组之间以及传统冬眠合剂组与改良冬眠合剂组之间比较，差异均无统计学意义（ P>0.05）。伤后3 d，相对于假伤组，单纯烫伤组大鼠肝脏组织中可见肝细胞弥漫微泡性脂肪变性和空泡变性，小肠组织中可见绒毛间质明显疏松水肿，肺脏组织中可见大片区域肺泡间隔明显增宽；相对于单纯烫伤组，2个冬眠合剂组大鼠的肝细胞脂肪变性和空泡变性都有所减轻，肺泡壁增厚和肠绒毛间质水肿均得到缓解，且其中改良冬眠合剂组大鼠各脏器组织学表现更接近假伤组。结论传统冬眠合剂可能有更强的抑炎作用，而改良冬眠合剂具有更好的肝功能保护作用，但这2种冬眠合剂均可缓解严重烫伤大鼠肝脏、肺脏和小肠的组织病理学损害。对于肝脏、肺脏和小肠，改良冬眠合剂具有不亚于传统冬眠合剂的脏器保护作用。
- 烧伤 /
- 炎症 /
- 组织学 /
- 改良冬眠合剂 /
- 脏器损伤
Abstract: Objective To compare the effects of the modified lytic cocktail and the classic lytic cocktail on organ function of severely scalded rats. Methods The experimental study method was applied. Twenty-four about 10-week-old male Sprague-Dawley rats were assigned into sham injury group, scald alone group, classic lytic cocktail group, and modified lytic cocktail group according to the random number table, with 6 rats in each group. In scald alone group, classic lytic cocktail group, and modified lytic cocktail group, rats were subjected to a 30% total body surface area (TBSA) full-thickness scald on the back. Rats in sham injury group underwent a simulated injury process to mimic a sham injury. Immediately after injury, rats in classic lytic cocktail group were intraperitoneally injected with a classic lytic cocktail (12 mL/kg) consisting of pethidine, chlorpromazine, and promethazine, supplemented with gavage using normal saline; and rats in modified lytic cocktail group were intraperitoneally injected with a mixed drug (2 mL/kg) consisting of midazolam and fentanyl, supplemented with gavage using cetirizine. Subsequently, rats in four groups were all intraperitoneally injected with lactated Ringer's solution for fluid resuscitation, with a total fluid and administration volume of 2 mL·kg ^-1·TBSA ^-1. On the following day, rats in the two lytic cocktail groups were administered medication once again as above. On post injury day (PID) 3, the abdominal aortic blood, liver, small intestine, and lung tissue were collected from rats in each group. The plasma levels of interleukin-1β (IL-1β), IL-10, and IL-6 were measured using an enzyme-linked immunosorbent assay. The plasma levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transferase (γ-GT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), LDH isoenzyme 1 (LDH-1), creatine kinase (CK), CK isoenzyme (CK-MB), urea, creatinine, and uric acid were detected using an automated biochemical analyzer. The histological changes of liver, small intestine, and lung tissue were observed after performing hematoxylin and eosin staining. Data were statistically analyzed with one-way analysis of variance and Tukey's test. Results On PID 3, the plasma level of IL-10 of rats in classic lytic cocktail group was (44±16) pg/mL, which was significantly higher than (20±9) pg/mL in modified lytic cocktail group and (21±6) pg/mL in scald alone group (with Pvalues all <0.05); there was no statistically significant difference in the plasma levels of IL-1β or IL-6 of rats among the four groups ( P>0.05). On PID 3, the plasma levels of ALT and AST of rats in scald alone group were (77±14) and (213±65) U/L, respectively, which were significantly higher than (59±5) and (108±10) U/L in sham injury group ( P<0.05); the plasma levels of ALT and AST in modified lytic cocktail group were (61±3) and (116±11) U/L, respectively, which were significantly lower than (81±13) and (207±54) U/L in classic lytic cocktail group ( P<0.05); the plasma level of AST of rats in modified lytic cocktail group was significantly lower than that in scald alone group ( P<0.05). On PID 3, there was no statistically significant difference in the plasma levels of γ-GT, ALP, LDH, LDH-1, CK, CK-MB, creatinine, or uric acid of rats among the four groups ( P>0.05); although there was a statistically significant overall difference in the plasma level of urea of rats among the four groups ( P<0.05), the comparisons between scald alone group and each of sham injury group, classic lytic cocktail group, and modified lytic cocktail group, and the comparison between classic lytic cocktail group and modified lytic cocktail group showed no statistically significant differences ( P>0.05). On PID 3, compared with those in sham injury group, rats in scald alone group exhibited diffuse microvesicular and vacuolar degeneration of hepatocytes in liver tissue, noticeable loose edema in the villous stroma in small intestine tissue, and significantly widened alveolar septa in large area of lung tissue. Compared with those in scald alone group, rats in the two lytic cocktail groups showed alleviated hepatocellular steatosis and vacuolar degeneration, relieved thickening of alveolar walls and edema in the villous stroma of the intestine. The histopathological manifestations of organs in rats of modified lytic cocktail group were closer to those in sham injury group. Conclusions The classic lytic cocktail may have a stronger anti-inflammatory effect, while the modified lytic cocktail exhibits better protection of liver function, but both of the two lytic cocktails can alleviate the histopathological injury of the liver, lungs, and small intestine in severely scalded rats. For the liver, lungs, and small intestine, the modified lytic cocktail provides organ protection comparable to that of the classic lytic cocktail.
- Burns /
- Inflammation /
- Histology /
- Modified lytic cocktail /
- Organ injury
于家傲：论文撰写、实验操作；王际壮：数据整理、研究指导；王家强：实验操作、数据整理；章雄、刘琰：论文修改、经费支持

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1 假伤组与3个严重烫伤组大鼠伤后3 d脏器组织学变化苏木精-伊红×200。1A、1B、1C、1D.分别为假伤组、单纯烫伤组、传统冬眠合剂组、改良冬眠合剂组肝脏组织，相对于图1A，图1B中可见肝细胞弥漫微泡性脂肪变性和空泡变性，肝窦明显淤血，图1C和1D中仅见少许微泡性脂肪变性及轻度空泡变性；1E、1F、1G、1H.分别为假伤组、单纯烫伤组、传统冬眠合剂组、改良冬眠合剂组小肠组织，相对于图1E，图1F中可见绒毛间质明显疏松水肿，图1G中可见绒毛间质轻度水肿，图1H中小肠组织形态无明显变化；1I、1J、1K、1L.分别为假伤组、单纯烫伤组、传统冬眠合剂组、改良冬眠合剂组肺脏组织，相对于图1I，图1J中可见大片区域肺泡间隔明显增宽，血管扩张充血，部分肺泡扩张，图1K部分区域肺泡间隔轻度增宽，血管轻度扩张充血，图1L中肺脏组织形态结构基本无明显变化

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表1 假伤组与3个严重烫伤组大鼠伤后3 d血浆中3种炎症指标水平比较（pg/mL， x ¯ ± s ）

表1. Comparison of the plasma levels of three inflammatory markers of rats in sham injury group and three severely scalded groups on post injury day 3

组别	样本数	IL-10	IL-6	IL-1β
假伤组	6	22±13	30±13	22±7
单纯烫伤组	6	21±6	55±12	37±17
传统冬眠合剂组	6	44±16	69±27	32±25
改良冬眠合剂组	6	20±9	36±10	33±13
F值		5.65	1.39	0.44
P值		0.007	0.276	0.730
P ₁值		0.998	0.651	0.697
P ₂值		0.021	0.921	0.979
P ₃值		0.999	0.781	0.992
P ₄值		0.011	0.414	>0.999
注：IL为白细胞介素； F值、 P值为4组间各指标总体比较所得， P ₁值为假伤组与单纯烫伤组比较所得， P ₂值、 P ₃值分别为单纯烫伤组与传统冬眠合剂组、改良冬眠合剂组比较所得， P ₄值为传统冬眠合剂组与改良冬眠合剂组比较所得

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表2 假伤组与3个严重烫伤组大鼠伤后3 d血浆中11种生化指标水平比较（ x ¯ ± s ）

表2. Comparison of the plasma levels of 11 biochemical markers of rats in sham injury group and three severely scalded groups on post injury day 3

组别	样本数	ALT（U/L）	AST（U/L）	ALP（U/L）	γ-GT（U/L）	肌酸激酶（U/L）
假伤组	6	59±5	108±10	2.2±0.6	0.41±0.17	220±51
单纯烫伤组	6	77±14	213±65	2.6±0.6	0.89±0.20	266±84
传统冬眠合剂组	6	81±13	207±54	3.4±0.6	0.73±0.25	214±121
改良冬眠合剂组	6	61±3	116±11	3.2±1.1	0.50±0.26	172±76
F值		7.85	9.80	2.92	0.92	1.21
P值		0.001	<0.001	0.061	0.449	0.332
P ₁值		0.018	0.003	0.804	0.458	0.785
P ₂值		0.919	0.943	0.361	0.958	0.720
P ₃值		0.089	0.006	0.667	0.631	0.259
P ₄值		0.020	0.008	0.943	0.893	0.832
注：ALT为丙氨酸转氨酶、AST为天冬氨酸转氨酶、γ-GT为γ-谷氨酰转移酶、ALP为碱性磷酸酶、LDH为乳酸脱氢酶、LDH-1为LDH同工酶1、CK-MB为肌酸激酶同工酶； F值、 P值为4组间各指标总体比较所得， P ₁值为假伤组与单纯烫伤组比较所得， P ₂值、 P ₃值分别为单纯烫伤组与传统冬眠合剂组、改良冬眠合剂组比较所得， P ₄值为传统冬眠合剂组与改良冬眠合剂组比较所得

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