不同劑量胰島素對嚴重燙傷大鼠早期血清IL

1、不同劑量胰島素對嚴重燙傷大鼠早期血清IL         07-08-24 11:10:00     編輯：studa20           作者：王耘川，高峰，賈赤宇，張萬福，湯朝武，胡大海 【關(guān)鍵詞】  燒傷；胰島素；炎癥；細胞因子類Regulating effects of different doses of insulin on serum IL1 and

2、 IL10 in early stage in severely scalded rats【Abstract】 AIM:  To determine the possible effects of different doses of insulin on the proinflammatory cytokine IL1 and the antiinflammatory cytokine IL10 in early stage of severe burn in rats. METHODS: Male SpragueDawly rats (30% TBSA, full thickne

3、ss burn) were randomly divided into 2 groups to receive either normal saline only (n=14, with intraperitoneal injection of 40 mL/kg normal saline, group A) or normal saline plus different doses of insulin (n=42, with intraperitoneal injection of 40 mL/kg normal saline plus subcutaneous injection of

4、1, 3 or 5 IU/kg insulin respectively, group B1, B2, B3). In addition, 7 rats without thermal injury were subjected to normal control group (group N). The outcome measures encompassed the serum glucose, blood pressure, serum IL1 and IL10 in rats at 6 and 24 h after scald and different treatments. RES

5、ULTS: The serum glucose increased quickly in rats after scald, which was demonstrated in group A to a range of (10.7±1.8) mmol/L by 6 h postscald. Insulin decreased the serum glucose in a dosedependent manner in rats of group B1, B2 and B3. And insulin at a dose of 3 IU/kg body weight reduced t

6、he serum glucose to a range of (6.0±0.8) mmol/L in rats at 6 h after thermal injury, which was almost eqeal to the normal range of (5.1±0.6) mmol/L (P>0.05). Meanwhile, insulin at this dose improved the blood pressure of scalded rats by keeping the blood pressure at a stable condition s

7、imilar to the normal. Insulin significantly decreased the serum IL1 in a dosedependent manner in scalded rats. It was partially showed after administration of insulin of 3 IU/kg  serum IL1  was  (54.0±8.6) ng/L and (46.8±6.8) ng/L, much lower than that without insulin admini

8、stration in rats at 6 and 24 h after thermal injury (P<0.01). Insulin could increase the serum IL10 in scalded rats. In comparison with group A, the most significant increase of serum IL10 was seen by application of 3 IU/kg insulin as (45.5±5.2) ng/L in rats 24 h after scald (P<0.01 vs gr

9、oup A). CONCLUSION:  Our data provided an insight that insulin might attenuate the inflammatory cascade response by decreasing the proinflammatory cytokine IL1 and increasing the antiinflammatory cytokine IL10, thereby keeping or restoring the systemic homeostasis after severe burn. The best ad

10、ministration dose of insulin in early burn stage should be determined by normalizing serum glucose.【Keywords】 burns; insulin; inflammation; cytokines【摘要】  目的： 研究不同劑量胰島素對嚴重燙傷早期血清IL1及IL10的影響. 方法： 將雄性SD大鼠63只隨機分為燙傷后鹽水治療（A）組、鹽水 + 不同劑量胰島素治療（B1B3）組，每組14只，另取7只正常大鼠作為對照（N）組. 燙傷各組在背部造成約30%體表總面積（TBSA）全層皮膚燙

11、傷. 傷后6，24 h后檢測血糖、血壓及血清IL1及IL10含量. 結(jié)果： 傷后大鼠血糖明顯升高，6 h達到（10.7±1.8）mmol/L，并持續(xù)高血糖狀態(tài)，胰島素可以降低傷后血糖，并隨劑量的增大而作用增強，3 IU/kg胰島素的6 h血糖為(6.0±0.8）mmol/L，與正常值（5.1±0.6）mmol/L比較無統(tǒng)計學(xué)差異（P>0.05）；傷后6，24 h血壓均明顯低于正常值，3 IU/kg胰島素對傷后血壓有顯著改善作用. 燙傷后血清IL1顯著升高，不同劑量胰島素干預(yù)均可降低IL1水平(P<0.05或P<0.01），3 IU/kg時血清IL

12、1在6, 24 h分別為（54.0±8.6），（46.8±6.8）ng/L. 燙傷后血清IL10含量有所升高，胰島素可進一步提高燙傷大鼠血清IL10含量，以3 IU/kg傷后24 h最為明顯，達到（45.5±5.2）ng/L，與單純鹽水治療組相比差異非常顯著（P<0.01）. 結(jié)論： 嚴重燒傷早期時不同劑量胰島素干預(yù)可調(diào)節(jié)血清促炎/抗炎性細胞因子水平，減輕傷后機體的炎癥反應(yīng)，當血糖控制在正常范圍時其作用最為明顯. 【關(guān)鍵詞】 燒傷；胰島素；炎癥；細胞因子類0引言近年來研究發(fā)現(xiàn)胰島素作為一種激素除了調(diào)節(jié)正常機體代謝外，還能對機體嚴重病患下的重要臟器功

13、能發(fā)揮保護作用，減輕失控性炎癥反應(yīng)及多器官功能損傷，提高患者生存率及后期生存質(zhì)量1. 嚴重燒傷后休克期機體處于持續(xù)性高血糖，并發(fā)失控性炎癥反應(yīng)及易于發(fā)生多器官功能衰竭等嚴重病變，我們擬通過在嚴重燒傷大鼠模型上早期應(yīng)用胰島素干預(yù)，觀察血糖、血壓及血清中促炎性細胞因子IL1及抗炎性細胞因子IL10水平的變化，探索不同胰島素劑量對炎癥反應(yīng)的調(diào)控及對休克的改善作用.1材料和方法1.1材料SureStep Plus血糖儀（美國lifescan公司）；BMP型生理壓力檢測儀（中國成都儀器廠）；Model 680型酶標儀（美國BIORAD公司）；精蛋白生物合成人胰島素注射液（丹麥Novo Nordisk公司

14、）；IL1，IL10 ELISA試劑盒（美國Biosource International公司）；SD大鼠63只（由第四軍醫(yī)大學(xué)實驗動物中心提供），雄性，體質(zhì)量180220 g. 1.2方法 1.2.1動物分組及制模將大鼠隨機分為燒傷后鹽水治療（A）組、鹽水+胰島素治療B1B3組（分別給于1， 3， 5 U/kg胰島素）組（每組14只）及對照（N）組7只. 30 g/L戊巴比妥鈉溶液40 mg/kg腹腔注射麻醉后，固定于燙傷專用模具上，浸入（94±1）水中20 s，造成大鼠背部約30%體表總面積（TBSA）全層皮膚燙傷（經(jīng)病理切片證實）. 傷后按40 mL/kg 生理鹽水立

15、即行腹腔補液，胰島素于左后肢腹側(cè)面行皮下注射. 在處死大鼠前1 h行頸外動脈插管，接生理壓力監(jiān)測儀監(jiān)測血壓變化. 隨后經(jīng)插管抽血處死大鼠，少量血用于檢測血糖，其余血液離心（1000 g，15 min），取上清液待用. 1.2.2血清IL1，IL10測定用ELISA試劑盒對標本中IL1，IL10含量進行測定. 待測品孔每孔加入待測血清樣品100 L；將反應(yīng)板充分混勻后置37，120 min；洗滌液充分洗滌46次，濾紙印干；每孔加入一抗工作液50 L，置37，60 min；洗滌同前；每孔加入酶標抗體工作液100 L，孵育和洗滌同前；每孔加入底物工作液100 L，置于37 暗處反應(yīng)10 min；每孔加入2 mol/L H2SO4 各1滴終止反應(yīng)；測A492 nm值. 實驗同時測定標準品，繪制標準曲線，根據(jù)標準曲線分別計算出待檢樣品中IL1，IL10的含量. 統(tǒng)計學(xué)處理： 數(shù)據(jù)以x±s表示，采用SPSS 11.5統(tǒng)計軟件進行數(shù)據(jù)處理，檢驗水準= 0.05.2結(jié)果2.1大鼠存活情況燙傷后24 h內(nèi)，N，A，B1，B2各組全部存活，B3組中有2只大鼠分別于傷后9，11 h死亡（29%）. 2.2血糖變化大鼠燙傷后血糖迅速升高， A組6，24 h血糖較正常值顯著升高（P<0.01）. 應(yīng)用胰島素干預(yù)后，B1，B2，B3各組血糖均有所