CAJ | 학술논문

目的观察重组白细胞介素10(IL-10)对大肠杆菌内毒素(LPS)诱导的实验性葡萄膜炎(EIU)的治疗作用.方法雄性Wistar大鼠56只随机分为3组,其中IL-10治疗组和阳性对照组各24只,正常对照组8只.治疗组和阳性对照组大鼠用LPS诱导EIU模型,治疗组24只大鼠在注射LPS前4.0、0.5 h分别按1μg/kg的剂量经尾静脉注射重组IL-10各1次,正常对照组大鼠同一部位注射等量生理盐水.阳性对照组和正常对照组大鼠用于对比观察.注射LPS后观察大鼠的发病开始时间和眼部表现,分别在注射LPS后4、24 h和3 d等3个时间点分批处理8只大鼠后取血和房水,检测血清和房水中肿瘤坏死因子(TNF)α、IL-6和IL-10水平,并进行病理检查和分级.结果阳性对照组24只大鼠全部出现葡萄膜炎反应,平均发病开始时间为(3.81±1.05)h,平均炎症评分为3.67±1.97,平均组织病理学分级为3.08±1.77.治疗组24只大鼠也全部出现轻度葡萄膜炎反应,平均发病开始时间为(5.63±1.02)h,平均炎症评分为2.00±1.25,平均组织病理学分级为1.67±1.17.治疗组大鼠的发病开始时间明显迟于阳性对照组大鼠(t=4.95,P=0.00),炎症评分(t=3.50,P=0.00)和组织病理学分级(t=3.28,P=0.00)也显著低于阳性对照组.正常对照组8只大鼠始终无阳性体征及病理改变.阳性对照组大鼠血清、房水TNF-α和IL-6水平明显高于治疗组和正常对照组(F=15.34,57.65,67.59,8.42;P=0.00),治疗组大鼠血清和房水IL-10水平明显高于阳性对照组和正常对照组(F=17.84,7.76;P=0.00).房水TNF-α、房水和血清IL-6与眼部炎症程度呈正相关(眼部表现评分r=0.58,0.31,0.81;组织病理学分级r=0.56,0.31,0.74;P=0.00);血清IL-10与炎症程度呈负相关(眼部评分和组织病理学分级分别为r=-0.54,-0.55;P=0.00),与发病开始时间呈正相关(r=0.73,P=0.00);血清及房水TNF-α和IL-6水平与发病开始时间呈负相关(r=-0.47,-0.59,-0.77,-0.36;P＜0.05).结论 IL-10能显著抑制LPS诱导的EIU模型中前炎症因子TNF-α和IL-6产生,减轻EIU眼部表现和病理损害,具有治疗EIU的作用. 目的觀察重組白細胞介素10(IL-10)對大腸桿菌內毒素(LPS)誘導的實驗性葡萄膜炎(EIU)的治療作用.方法雄性Wistar大鼠56隻隨機分為3組,其中IL-10治療組和暘性對照組各24隻,正常對照組8隻.治療組和暘性對照組大鼠用LPS誘導EIU模型,治療組24隻大鼠在註射LPS前4.0、0.5 h分彆按1μg/kg的劑量經尾靜脈註射重組IL-10各1次,正常對照組大鼠同一部位註射等量生理鹽水.暘性對照組和正常對照組大鼠用于對比觀察.註射LPS後觀察大鼠的髮病開始時間和眼部錶現,分彆在註射LPS後4、24 h和3 d等3箇時間點分批處理8隻大鼠後取血和房水,檢測血清和房水中腫瘤壞死因子(TNF)α、IL-6和IL-10水平,併進行病理檢查和分級.結果暘性對照組24隻大鼠全部齣現葡萄膜炎反應,平均髮病開始時間為(3.81±1.05)h,平均炎癥評分為3.67±1.97,平均組織病理學分級為3.08±1.77.治療組24隻大鼠也全部齣現輕度葡萄膜炎反應,平均髮病開始時間為(5.63±1.02)h,平均炎癥評分為2.00±1.25,平均組織病理學分級為1.67±1.17.治療組大鼠的髮病開始時間明顯遲于暘性對照組大鼠(t=4.95,P=0.00),炎癥評分(t=3.50,P=0.00)和組織病理學分級(t=3.28,P=0.00)也顯著低于暘性對照組.正常對照組8隻大鼠始終無暘性體徵及病理改變.暘性對照組大鼠血清、房水TNF-α和IL-6水平明顯高于治療組和正常對照組(F=15.34,57.65,67.59,8.42;P=0.00),治療組大鼠血清和房水IL-10水平明顯高于暘性對照組和正常對照組(F=17.84,7.76;P=0.00).房水TNF-α、房水和血清IL-6與眼部炎癥程度呈正相關(眼部錶現評分r=0.58,0.31,0.81;組織病理學分級r=0.56,0.31,0.74;P=0.00);血清IL-10與炎癥程度呈負相關(眼部評分和組織病理學分級分彆為r=-0.54,-0.55;P=0.00),與髮病開始時間呈正相關(r=0.73,P=0.00);血清及房水TNF-α和IL-6水平與髮病開始時間呈負相關(r=-0.47,-0.59,-0.77,-0.36;P＜0.05).結論 IL-10能顯著抑製LPS誘導的EIU模型中前炎癥因子TNF-α和IL-6產生,減輕EIU眼部錶現和病理損害,具有治療EIU的作用.
목적 관찰중조백세포개소10(IL-10)대대장간균내독소(LPS)유도적실험성포도막염(EIU)적치료작용.방법 웅성Wistar대서56지수궤분위3조,기중IL-10치료조화양성대조조각24지,정상대조조8지.치료조화양성대조조대서용LPS유도EIU모형,치료조24지대서재주사LPS전4.0、0.5 h분별안1μg/kg적제량경미정맥주사중조IL-10각1차,정상대조조대서동일부위주사등량생리염수.양성대조조화정상대조조대서용우대비관찰.주사LPS후관찰대서적발병개시시간화안부표현,분별재주사LPS후4、24 h화3 d등3개시간점분비처리8지대서후취혈화방수,검측혈청화방수중종류배사인자(TNF)α、IL-6화IL-10수평,병진행병리검사화분급.결과 양성대조조24지대서전부출현포도막염반응,평균발병개시시간위(3.81±1.05)h,평균염증평분위3.67±1.97,평균조직병이학분급위3.08±1.77.치료조24지대서야전부출현경도포도막염반응,평균발병개시시간위(5.63±1.02)h,평균염증평분위2.00±1.25,평균조직병이학분급위1.67±1.17.치료조대서적발병개시시간명현지우양성대조조대서(t=4.95,P=0.00),염증평분(t=3.50,P=0.00)화조직병이학분급(t=3.28,P=0.00)야현저저우양성대조조.정상대조조8지대서시종무양성체정급병리개변.양성대조조대서혈청、방수TNF-α화IL-6수평명현고우치료조화정상대조조(F=15.34,57.65,67.59,8.42;P=0.00),치료조대서혈청화방수IL-10수평명현고우양성대조조화정상대조조(F=17.84,7.76;P=0.00).방수TNF-α、방수화혈청IL-6여안부염증정도정정상관(안부표현평분r=0.58,0.31,0.81;조직병이학분급r=0.56,0.31,0.74;P=0.00);혈청IL-10여염증정도정부상관(안부평분화조직병이학분급분별위r=-0.54,-0.55;P=0.00),여발병개시시간정정상관(r=0.73,P=0.00);혈청급방수TNF-α화IL-6수평여발병개시시간정부상관(r=-0.47,-0.59,-0.77,-0.36;P＜0.05).결론 IL-10능현저억제LPS유도적EIU모형중전염증인자TNF-α화IL-6산생,감경EIU안부표현화병리손해,구유치료EIU적작용.
Objective To examine the role of recombinant interleukin-10 (IL-10) and the therapeutic effect of endotoxin-induced uveitis (EIU) in rats. Methods Fifty-six male Wistar rats were randomized into three groups. IL-10 treatment group and positive control group had 24 rats respectively, and the normal control group had eight rats. Endotoxin-induced uveitis (EIU) is an established animal model of acute ocular inflammation induced by LPS intravenous injection ( 1 μg/kg). The onset times and signs were observed and the clinical scores were recorded. The blood samples and the aqueous humor samples of right eye were collected separately before the rats were sacrificed at fourth hour, 24th hour and third day after LPS injection. The enzyme-linked immunosorbent assay was used to measure tumor necrosis factor (TNF) α,IL-6, and IL-10 levels in the serum and aqueous humor. The left eyes were used for pathological examination and pathological grading. Results The symptoms of uveitis were appeared in all 24 rats in the positive group. The average onset time was (3.81 ± 1.05) hours, the average clinical score was 3.67±l. 97. The mild manifestations of uveitis were also appeared in all of the rats in treatment group. The average onset time was (5. 63±1.02) hours, the average clinical score was 2.00± 1.25. The average onset time in treatment group was postponed compared with the rats of positive group (t=4.95, P=0. 000).The clinical scores (t= 3. 50, P= 0. 00) and the pathological grades (t= 3.28, P= 0. 00) in treatment group were lower than those of positive group. There were not signs or pathologic changes in all the eight rats in the negative control group. The serum and aqueous humor levels of TNF-α and IL-6 in the rats of positive group were higher than those of the treatment group and control group (F=15.34, 57.65, 67.59, 8.42;P=0. 00). The serum and aqueous humor levels of IL-10 in the rats of treatment group were higher than those of the positive group and the control group (F = 17.84, 7.76; P = 0.00). There were positive correlations between the level of aqueous humor TNF-α, serum and aqueous humor levels of IL-6 and the disease severity (reye=0. 58, 0. 31, 0. 81, rpath =0. 56, 0. 31, 0. 74; P＜0.05). The negative correlations were presented between the serum levels of IL-10 with the disease severity (r=- 0. 54,-0. 55; P= 0.00).There were negative correlations between the serum and aqueous humor levels of TNF-α and IL-6 and the onset timeof the disease (r=-0.47,-0.59,-0.77,-0.36; P＜0.05) as well. Conclusions These findings strongly suggest that suppressive IL-10 is a potent candidate for the prevention of TNF-α and IL-6 in uveitis and could be applied as a novel immunoregulatory agent to control EIU.