白细胞介素-10对大鼠重症急性胰腺炎血清肿瘤坏死因子-α及胰腺组织的影响

Effects of IL-10 on serum TNF-α and pancreatic tissue in rats with severe acute pancreatitis

目的探讨重组人白细胞介素-10（IL-10）对大鼠重症急性胰腺炎（SAP）血清肿瘤坏死因子（TNF）-α及胰腺组织病理改变的影响，为临床治疗SAP提供相关理论依据。方法健康雄性SD大鼠90只，随机分为空白对照组（N组）30只，SAP组（S组）30只，IL-10干预组（Ⅰ组）30只。S组和Ⅰ组采用5％牛磺胆酸钠逆行胰胆管注射方法制作SAP模型，N组开腹后翻动胰腺即关腹。Ⅰ组分别在术后第1、3、5h腹腔注射重组人IL-1010000U，N组和A组大鼠分别在相同时间点腹腔注射等量生理盐水。3组分别在6、12、24h分批处死大鼠，用酶联免疫吸附法（ELISA）检测血清TNF—α，生化法检测血清淀粉酶水平，HE染色对胰腺组织进行病理学评分。结果S组6、12、24h血清淀粉酶分别为（6633．9±846．7）、（9421．4±1031．8）、（8755．6±734．5）IU／L、血清TNF-α水平为（87．6±3．3）、（113．3±10．2）、（100．2±2．3）ng／L，胰腺组织病理评分为8．68±0．63、13．41±0．79、16．78±0．87，较N组各时间点血清淀粉酶（1025．3±326．9）、（999．9±212．3）、（962．3±128．9）IU／L、血清TNF-α水平[（55．6±2．1）、（56．1±2．2）、（58．7±1．3）ng／L]，胰腺组织病理评分（0．13±0．11、0．15±0．12、0．16±0．15）明显增高（P〈0．05）。其中，胰腺组织病理学评分以24h最为显著，血清淀粉酶和TNF-α水平在12h达到高峰。Ⅰ组6、12、24h胰腺组织病理学评分为6．52±0．54、9．37±0．35、12．43±0．69、血清淀粉酶为（6032．8±534．9）、（7475．8±834．2）、（6903．4±377．1）IU／L，TNF-α水平为（67．5±2．5）、（93．0±4．9）、（86．7±6．6）ng／L，在各时点均显著低于S组（P〈0．05）。结论SAP早期应用IL-10可以抑制炎症因子TNF-α释放，降低炎症反应，改善胰腺组织病理病变。

Objective To investigate the effects of recombinant human interleukin-10 （IL-10） on serum tumor necrosis factor α （TNF-α） and histopathological changes of pancreas in rats with severe acute pancreatitis （SAP）, and provide theorical basis for SAP clinical treatment. Methods 90 Sprague-Dawley （SD） rats were randomly divided into three groups： normal control group （group N, n = 30）, SAP group （group S, n = 30） and IL-10 interference group （group I, n = 30）. 5% sodium taurocholate was retrogradely injected into the pancreatic duct in S group and I group to induce SAP model. Rats in N group whose pancreas was just flipped and stricken gently without injection. Group I was treated with 10 000 units of intraperitioneal recombinant human IL-10 at 1, 3 and 5 h. Group N and group S received three intraperitoneal injections of 0.9% normal saline at the same time points. Rats were killed at 6, 12 and 24 h. The level of TNF-α in serum was determined by enzyme-linked immunosorbent assay （ELISA） , and the level of amylase was assayed by biochemical methods. The pancreas histological changes were observed by H-E staining. Results Compared with group N [ serum amylase （ 1 025.3 ± 326.9 ）, （ 999.9 ± 212.3 ）, （ 962.3 ± 128.9） IU/L; TNF-α （55.6 ± 2. 1 ）, （56. 1 ±2.2）, （58.7 ± 1.3）ng/L; pancreatic histopathological score 0.13 ± 0.11,0. 15 ± 0.12, 0.16 ± 0.151 , serum amylase （6 633.9 ± 846.7）, （9 421.4 ± 1 031.8）, （8755.6±734.5） IU/L, TNF-α （87.6 ±3.3）, （113.3 ±10.2）, （100.2 ±2.3） ng/L and pancreatic histopathological score （8.58 ±0. 63, 13.41 ±0.79, 16.78 ±0.87） in group S were increased significantlyat 6, 12 and 24 h （P 〈0. 05）. The pancreatic damage at 24 h was the most severe, and the peak concentration of AMY and TNF-α reached at 12 h. Compared with group S, pancreatic histopathological scores 6.52 ± 0.54, 9.37 ±0. 35, 12.43 ± 0.69, the level of serum amylase （6 032.8 ± 534.9 ）, （7 475.8 ± 834.2 ）, （ 6 903.4 ± 377.1 ） IU/L and TNF-α （ 67.5 ± 2.5 ）, （ 93.0 ± 4.9 ）, （ 86.7 ± 6.6 ） ng/L in group I were significantly decreased at corresponding time points （ P 〈 0.05 ）. Cortelusions Early application of recombinant human IL-10 can attenuate SAP inflammatory response and relieve the histopathological injury of pancreas by inhibiting the release of TNF-α. IL-10 can be used for the treatment of SAP.