重症胰腺炎Th1/Th2型细胞因子变化及乌司他丁影响的实验研究
摘要
目的:重症急性胰腺炎(Sever acute pancreatitis,SAP)以其起病急,进展快,病情凶险为特点,早期即可并发多器官功能衰竭,虽然目前治疗措施较多,但死亡率仍然较高,其发病机制尚不完全清楚。本实验通过逆行胰胆管注射5%牛磺胆酸钠制成SAP模型,研究在SAP模型中以及施加乌司他丁干预后Th1/Th2型细胞因子在外周血中的变化规律,探讨Th1/Th2型细胞因子与SAP严重程度和预后的关系,以及应用乌司他丁治疗SAP对Th1/Th2型细胞因子的影响。
方法:选取Wistar大鼠112只,均为雄性,体重240±30g,随机分为16组,每组7只,即正常对照组(A组),假手术组(B组),SAP组(C组)和乌司他丁干预组(D组),其中B组分为2h,6h,12h,24h处死组和生存组(分别记为B1,B2,B3,B4和B生存),同样C组和D组亦分为2h,6h,12h,24h处死组和生存组(分别记为C1,C2,C3,C4,C生存和D1,D2,D3,D4,D生存)。SAP组用3.5%的水合氯醛(0.8ml/100g)行腹腔注射麻醉后,仰卧固定于实验台上,备皮,消毒,铺巾,作剑突下正中切口长约1.5cm,入腹后找到十二指肠将其提起,可见胰胆管及十二指肠乳头,在十二指肠乳头对侧肠壁用皮试针扎一小孔,再用特制的带金属头的钝头5号针头自该孔进入十二指肠肠腔,轻柔的穿过十二指肠乳头进入胰胆管约1cm,在靠近十二指肠用2-0号带线缝针缝扎胰胆管,打活结固定5号针头,在靠近肝门处用一血管夹夹闭胰胆管近心端,用1ml皮试空针匀速逆行胰胆管内注射5%牛磺胆酸钠(0.1ml/100g,0.1ml/min),注射完毕后保持8分钟,拔出针头,松开血管夹和结扎线,腹腔内留置青霉素20万单位,皮下注射生理盐水补液(3ml/100g),做好标记放入鼠笼自由进食水,按规定时间(术后2h,6h,12h,24h)行心脏采血并处死大鼠,观察腹内脏器情况,收集腹水和胰腺头部,C生存用于观察生存期,所采全血37℃水浴半小时后离心3000rpm×10min,用EP管收集血清1.5ml冻存于-70℃冰箱待用,测量腹水后用EP管装1.5ml放入-70℃冰箱待用,所采组织标本放入4%多聚甲醛固定,交技术人员进行脱水,包埋,切片,HE染色。干预组D组在麻醉后立即臀部皮下注射乌司他丁(3000u/100g),其余步骤同C组。B组开腹后仅翻动胃十二指肠数次,然后关腹,其余步骤同C组。用ELISA法检测血清细胞因子IFN-γ和IL-4,分别代表Th1和Th2型细胞因子,检测腹水和血清淀粉酶,光镜下观察胰腺组织病理变化,评估其病变程度,观察B生存,C生存和D生存各组生存期。
结果:①B组各时间点无腹水,血清淀粉酶和Th1,Th2型细胞因子及Th1/Th2比值无明显变化,光镜下见胰腺组织无水肿出血坏死及炎症细胞浸润,生存期组在两周内无死亡。②C组血性腹水量随时间的增加而增加,在24h达到高峰(2h,6h,12h之间P<0.05,但24h与12h比较P=0.059>0.05),腹水淀粉酶随时间的增加而增加(相邻时间点比较P<0.05),血清淀粉酶随时间的增加而增加(2h,6h,12h相邻两者之间P<0.05,但24h与12h比较P=0.79>0.05),Th1型细胞因子在2h较A组有轻度的升高(P<0.05),但随即随时间的增加而降低(P<0.05),Th2型细胞因子在2h较A组轻度升高(P=0.18>0.05),之后随时间的增加而显著升高(P<0.05),Th1/Th2比值在2h较A组轻度升高(P=0.082>0.05),之后其比值迅速降低,甚至出现比例倒置。光镜下见胰腺水肿、出血、坏死和炎细胞浸润,其严重程度病理评分随时间的增加而增加(2h,6h,12h相邻两者之间P<0.05,但24h与12h比较P=0.091>0.05),生存期组存活2.14±1.07天;③D组血性腹水量随时间的增加而增加(P<0.05),腹水淀粉酶和血清淀粉酶均随时间的增加而增加(P<0.05),Th1型细胞因子在2h较A组有轻度的升高(P<0.05),但随即随时间的增加而下降(P<0.05),Th2型细胞因子随时间的增加而升高(2h,6h,12h相邻两者之间P>0.05,但24h与12h比较P<0.05),Th1/Th2比值在2h,6h随时间的增加其比值升高(P<0.05),之后随时间的增加其比值降低(P<0.05),光镜下见胰腺水肿、出血、坏死和炎细胞浸润,其病理评分随时间的增加而增加(2h,6h,12h相邻两者之间P<0.05,24h与12h比较P>0.05),生存期组存活5.86±2.41天;④相同时点D组与C组比较: D组腹水量,腹水淀粉酶,血清淀粉酶和胰腺病理评分均较C组减轻,D生存较C生存延长(P<0.05),Th1型细胞因子升高(P<0.05),Th2型细胞因子降低(2h,P>0.05,其余P<0.05),Th1/Th2比值显著升高(P<0.05),不同程度逆转了Th1/Th2的极化。
结论:1、SAP早期,总体上随着病情的加重Th1型细胞因子逐渐降低, Th2型细胞因子逐渐升高,Th1/Th2比值逐渐降低,趋向于Th1抑制和Th2极化。
2、Th1/Th2的失衡程度与SAP的严重程度有关,纠正Th1/Th2的失衡状态,可能在减缓SAP疾病的演进过程中有积极意义。
3、检测Th1/Th2型细胞因子可能对SAP预后的评估有重要意义。
Objective: SAP(Sever acute pancreatitis) is characteristic of acute onset,fast progression and dangerous pathogenetic condition, while multiple organ failure may happen early. Though many therapeutic measures are taken now , the death rate is still much high, and the pathogenesy of SAP is not distinct completely. SAP model was induced by retrograde injection of 5% sodium taurocholate to pancreatic duct. we detected the concentration change of Th1/Th2 cytokines in peripheral blood.,and studied the relation between Th1/Th2 cytokines and the severity degree and prognosis of SAP, as well as investigated the effect of Ulinastatin to Th1/Th2 cytokines.
Methods: 112 health Wistar rat(all maleness, body weight 240±30g) were divided into 16 groups randomly,7 per group,that is:norm control group(A group),sham operation group (B group),SAP group(C group) and Ulinastatin intervention group(D group). B group was divided into 2h,6h,12h,24h groups and survival groups(marked with B_1,B_2,B_3,B_4 and B_(sur) respectively), likewise, C group and D group were divided into C_1,C_2,C_3,C_4,C_(sur) and D_1,D_2,D_3,D_4,D_(sur).SAP model was induced by retrograde injection of 5% sodium tanrocholate to pancreatic duct. Compared with SAP group , stomach and duodenum were flipped only and 5% sodium tanrocholate was not injected in sham operation group ,as well as ulinastatin was used immediately after anesthesia in Ulinastatin intervention group.Nothing was done in norm control group.Then, at 2h,6h,12h,24h the blood and pancreatic tissue were collected to detect Th1/Th2 cytokines by ELISA and necrosis grade by HE stainning. The quantity of ascites was measured , the amylase of blood and ascetes was detected and the life span of every survival groups was observed.
Results:①At every time point,B group had no ascites, serum amylase,Th1,Th2 cytokines and the ratio of Th1/Th2 had no significant changes. Pancreatitic tissue had no edema, hemorrhage, necrosis and inflammatory cell infiltration by light microscope. And there were no death in postoperation two weeks.②C group: The quantity of hemorrhagic ascites increased gradually with time’s passing and reached the peak at 24h(P<0.05 among 2h,6h,12h,but 24h compared with 12h,P=0.059>0.05),and so did ascetic amylase(P<0.05)and serum amylase(P<0.05 among 2h,6h,12h,but 24h compared with 12h, P=0.79>0.05).Th1 cytokines increased temperately compared with A group at 2h(P<0.05),but immediately decreased with time’s passing(P < 0.05). Th2 cytokines increased temperately compared with A group at 2h(P=0.18>0.05),and after that, increased significantly(P < 0.05).The ratio of Th1/Th2 rose temperately compared with A group at 2h(P=0.082>0.05),then falled significantly with time’s passing, even the ratio was inverted. Pancreatic tissue was edema, hemorrhage, necrosis and inflammatory cell infiltration by light microscope, and its severity degree rose with time’s passing(P<0.05 among 2h,6h,12h,but P=0.091>0.05 at 24h compared with 12h),the life span of Csur group was 2.14±1.07d.③D group: The quantity of hemorrhagic ascites stepped up with time’s passing(P<0.05), and so did ascetic amylase and serum amylase(P<0.05). Th1 cytokines increased temperately compared with A group at 2h(P<0.05),but immediately falled with time’s passing(P<0.05), Th1 cytokines increased temperately with time’s passing,but there was no significantly difference among 2h,6h and 12h(P>0.05)except between 24h and 12h. The ratio of Th1/Th2 rose with time’s passing at 2h and 6h(P<0.05),then that was falled(P<0.05). Pancreatic tissue was edema, hemorrhage, necrosis and inflammatory cell infiltration by light microscope, and its severity degree rose with time’s passing(P>0.05 among 2h,6h,12h,but P=0.091<0.05 at 24h compared with 12h),the life span of Csur group was 5.86±2.41d.④At the same time,D group was compared with C group: the quantity of ascites, ascetic amylase , serum amylase and pancreatic patho-score were smaller and the life span of Dsur was longer(P<0.05).Th1 cytokines rose(P<0.05), however Th2 cytokines falled(P>0.05 at 2h,P<0.05 at the rest time points),and the ratio of Th1/Th2 rose significantly(P<0.05),the polarization was reverted partly.
Conclusion:1.With pathogenetic condition’s aggravation, Th1 cytokines fall,but Th2 cytokines increase gradually and the ratio of Th1/Th2 falls, inclined to Th1 suppression and Th2 polarization in early SAP.
2. The imbalance of Th1/Th2 is concerned with the severity of SAP and Correcting the imbalance of Th1/Th2 is maybe very valuable in lessening the progress of SAP.
3. It is much significant to detect Th1/Th2 cytokines in evaluating the prognosis of SAP.
方法:选取Wistar大鼠112只,均为雄性,体重240±30g,随机分为16组,每组7只,即正常对照组(A组),假手术组(B组),SAP组(C组)和乌司他丁干预组(D组),其中B组分为2h,6h,12h,24h处死组和生存组(分别记为B1,B2,B3,B4和B生存),同样C组和D组亦分为2h,6h,12h,24h处死组和生存组(分别记为C1,C2,C3,C4,C生存和D1,D2,D3,D4,D生存)。SAP组用3.5%的水合氯醛(0.8ml/100g)行腹腔注射麻醉后,仰卧固定于实验台上,备皮,消毒,铺巾,作剑突下正中切口长约1.5cm,入腹后找到十二指肠将其提起,可见胰胆管及十二指肠乳头,在十二指肠乳头对侧肠壁用皮试针扎一小孔,再用特制的带金属头的钝头5号针头自该孔进入十二指肠肠腔,轻柔的穿过十二指肠乳头进入胰胆管约1cm,在靠近十二指肠用2-0号带线缝针缝扎胰胆管,打活结固定5号针头,在靠近肝门处用一血管夹夹闭胰胆管近心端,用1ml皮试空针匀速逆行胰胆管内注射5%牛磺胆酸钠(0.1ml/100g,0.1ml/min),注射完毕后保持8分钟,拔出针头,松开血管夹和结扎线,腹腔内留置青霉素20万单位,皮下注射生理盐水补液(3ml/100g),做好标记放入鼠笼自由进食水,按规定时间(术后2h,6h,12h,24h)行心脏采血并处死大鼠,观察腹内脏器情况,收集腹水和胰腺头部,C生存用于观察生存期,所采全血37℃水浴半小时后离心3000rpm×10min,用EP管收集血清1.5ml冻存于-70℃冰箱待用,测量腹水后用EP管装1.5ml放入-70℃冰箱待用,所采组织标本放入4%多聚甲醛固定,交技术人员进行脱水,包埋,切片,HE染色。干预组D组在麻醉后立即臀部皮下注射乌司他丁(3000u/100g),其余步骤同C组。B组开腹后仅翻动胃十二指肠数次,然后关腹,其余步骤同C组。用ELISA法检测血清细胞因子IFN-γ和IL-4,分别代表Th1和Th2型细胞因子,检测腹水和血清淀粉酶,光镜下观察胰腺组织病理变化,评估其病变程度,观察B生存,C生存和D生存各组生存期。
结果:①B组各时间点无腹水,血清淀粉酶和Th1,Th2型细胞因子及Th1/Th2比值无明显变化,光镜下见胰腺组织无水肿出血坏死及炎症细胞浸润,生存期组在两周内无死亡。②C组血性腹水量随时间的增加而增加,在24h达到高峰(2h,6h,12h之间P<0.05,但24h与12h比较P=0.059>0.05),腹水淀粉酶随时间的增加而增加(相邻时间点比较P<0.05),血清淀粉酶随时间的增加而增加(2h,6h,12h相邻两者之间P<0.05,但24h与12h比较P=0.79>0.05),Th1型细胞因子在2h较A组有轻度的升高(P<0.05),但随即随时间的增加而降低(P<0.05),Th2型细胞因子在2h较A组轻度升高(P=0.18>0.05),之后随时间的增加而显著升高(P<0.05),Th1/Th2比值在2h较A组轻度升高(P=0.082>0.05),之后其比值迅速降低,甚至出现比例倒置。光镜下见胰腺水肿、出血、坏死和炎细胞浸润,其严重程度病理评分随时间的增加而增加(2h,6h,12h相邻两者之间P<0.05,但24h与12h比较P=0.091>0.05),生存期组存活2.14±1.07天;③D组血性腹水量随时间的增加而增加(P<0.05),腹水淀粉酶和血清淀粉酶均随时间的增加而增加(P<0.05),Th1型细胞因子在2h较A组有轻度的升高(P<0.05),但随即随时间的增加而下降(P<0.05),Th2型细胞因子随时间的增加而升高(2h,6h,12h相邻两者之间P>0.05,但24h与12h比较P<0.05),Th1/Th2比值在2h,6h随时间的增加其比值升高(P<0.05),之后随时间的增加其比值降低(P<0.05),光镜下见胰腺水肿、出血、坏死和炎细胞浸润,其病理评分随时间的增加而增加(2h,6h,12h相邻两者之间P<0.05,24h与12h比较P>0.05),生存期组存活5.86±2.41天;④相同时点D组与C组比较: D组腹水量,腹水淀粉酶,血清淀粉酶和胰腺病理评分均较C组减轻,D生存较C生存延长(P<0.05),Th1型细胞因子升高(P<0.05),Th2型细胞因子降低(2h,P>0.05,其余P<0.05),Th1/Th2比值显著升高(P<0.05),不同程度逆转了Th1/Th2的极化。
结论:1、SAP早期,总体上随着病情的加重Th1型细胞因子逐渐降低, Th2型细胞因子逐渐升高,Th1/Th2比值逐渐降低,趋向于Th1抑制和Th2极化。
2、Th1/Th2的失衡程度与SAP的严重程度有关,纠正Th1/Th2的失衡状态,可能在减缓SAP疾病的演进过程中有积极意义。
3、检测Th1/Th2型细胞因子可能对SAP预后的评估有重要意义。
Objective: SAP(Sever acute pancreatitis) is characteristic of acute onset,fast progression and dangerous pathogenetic condition, while multiple organ failure may happen early. Though many therapeutic measures are taken now , the death rate is still much high, and the pathogenesy of SAP is not distinct completely. SAP model was induced by retrograde injection of 5% sodium taurocholate to pancreatic duct. we detected the concentration change of Th1/Th2 cytokines in peripheral blood.,and studied the relation between Th1/Th2 cytokines and the severity degree and prognosis of SAP, as well as investigated the effect of Ulinastatin to Th1/Th2 cytokines.
Methods: 112 health Wistar rat(all maleness, body weight 240±30g) were divided into 16 groups randomly,7 per group,that is:norm control group(A group),sham operation group (B group),SAP group(C group) and Ulinastatin intervention group(D group). B group was divided into 2h,6h,12h,24h groups and survival groups(marked with B_1,B_2,B_3,B_4 and B_(sur) respectively), likewise, C group and D group were divided into C_1,C_2,C_3,C_4,C_(sur) and D_1,D_2,D_3,D_4,D_(sur).SAP model was induced by retrograde injection of 5% sodium tanrocholate to pancreatic duct. Compared with SAP group , stomach and duodenum were flipped only and 5% sodium tanrocholate was not injected in sham operation group ,as well as ulinastatin was used immediately after anesthesia in Ulinastatin intervention group.Nothing was done in norm control group.Then, at 2h,6h,12h,24h the blood and pancreatic tissue were collected to detect Th1/Th2 cytokines by ELISA and necrosis grade by HE stainning. The quantity of ascites was measured , the amylase of blood and ascetes was detected and the life span of every survival groups was observed.
Results:①At every time point,B group had no ascites, serum amylase,Th1,Th2 cytokines and the ratio of Th1/Th2 had no significant changes. Pancreatitic tissue had no edema, hemorrhage, necrosis and inflammatory cell infiltration by light microscope. And there were no death in postoperation two weeks.②C group: The quantity of hemorrhagic ascites increased gradually with time’s passing and reached the peak at 24h(P<0.05 among 2h,6h,12h,but 24h compared with 12h,P=0.059>0.05),and so did ascetic amylase(P<0.05)and serum amylase(P<0.05 among 2h,6h,12h,but 24h compared with 12h, P=0.79>0.05).Th1 cytokines increased temperately compared with A group at 2h(P<0.05),but immediately decreased with time’s passing(P < 0.05). Th2 cytokines increased temperately compared with A group at 2h(P=0.18>0.05),and after that, increased significantly(P < 0.05).The ratio of Th1/Th2 rose temperately compared with A group at 2h(P=0.082>0.05),then falled significantly with time’s passing, even the ratio was inverted. Pancreatic tissue was edema, hemorrhage, necrosis and inflammatory cell infiltration by light microscope, and its severity degree rose with time’s passing(P<0.05 among 2h,6h,12h,but P=0.091>0.05 at 24h compared with 12h),the life span of Csur group was 2.14±1.07d.③D group: The quantity of hemorrhagic ascites stepped up with time’s passing(P<0.05), and so did ascetic amylase and serum amylase(P<0.05). Th1 cytokines increased temperately compared with A group at 2h(P<0.05),but immediately falled with time’s passing(P<0.05), Th1 cytokines increased temperately with time’s passing,but there was no significantly difference among 2h,6h and 12h(P>0.05)except between 24h and 12h. The ratio of Th1/Th2 rose with time’s passing at 2h and 6h(P<0.05),then that was falled(P<0.05). Pancreatic tissue was edema, hemorrhage, necrosis and inflammatory cell infiltration by light microscope, and its severity degree rose with time’s passing(P>0.05 among 2h,6h,12h,but P=0.091<0.05 at 24h compared with 12h),the life span of Csur group was 5.86±2.41d.④At the same time,D group was compared with C group: the quantity of ascites, ascetic amylase , serum amylase and pancreatic patho-score were smaller and the life span of Dsur was longer(P<0.05).Th1 cytokines rose(P<0.05), however Th2 cytokines falled(P>0.05 at 2h,P<0.05 at the rest time points),and the ratio of Th1/Th2 rose significantly(P<0.05),the polarization was reverted partly.
Conclusion:1.With pathogenetic condition’s aggravation, Th1 cytokines fall,but Th2 cytokines increase gradually and the ratio of Th1/Th2 falls, inclined to Th1 suppression and Th2 polarization in early SAP.
2. The imbalance of Th1/Th2 is concerned with the severity of SAP and Correcting the imbalance of Th1/Th2 is maybe very valuable in lessening the progress of SAP.
3. It is much significant to detect Th1/Th2 cytokines in evaluating the prognosis of SAP.
引文
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[6] Bhatia M, Brady M, Shokubi S, et al. Inflammatory mediators in acute pancreatitis[J]. J Pathol, 2000,190(2):117-125
[7] Laser HG, Gross V, Scheibenbogen C, et al. Elevation of serum interleukin-6 concentration precedes acute phase response and reflects severity in acute pancreatitis [J] .Gastroenterology, 1991,101:782-785
[8] 安云庆,高晓明.医学免疫学[M].北京:北京大学医学出版社,2004:74-78
[9] Wrobleski DM, Barth MM,Oyen LJ .Necrotizing pancreatitis pathophysi- ology,diagnosis, and acute care management[J].AACN Clin Issues, 1999,10(4): 464-477.
[10] 吕云福.现代胰腺外科学[M].北京:人民军医出版社,2003:126-134
[11] Ueda T, Takeyama Y, Yasuda T, et al. Functional alterations of splenocytes in severe acute pancreatitis [J]. J Surg Res, 2002, 102(2) :161-168
[12] Ueda T ,Takeyama Y, Yasuda T, et al. Significant elevation of serum interleukin-18 levels in patients with acute pancreatitis[J].J Gastroenterol, 2006,41(2):158-165
[13] Richard AG. Immunology[M]. 5th ed. New York:Freeman and Company, 2003:288-291
[14] 江晓东,陈惠晓.Th 细胞的分化调控[J].国外医学·免疫学分册,2003,26(2): 79-82
[15] Ouyang W,Lohning M, Gao Z, et al. Stat6-Independent GATA-3 Autoactivation Directs IL-4-Independent Th2 Development and Commitment [J]. Immunity, 2000,12(1):27-37
[16] Dorfman DM, van den Elzen P, Weng AP, et al. Differential expression of T-bet,a T-box transcription factor required for Th1 T-cell development ,in perihheral T-cell lymphomas[J]. Am J Clin Pathol, 2003, 120(6): 866-873
[17] Rarcity MG,Connor S,Criddle CN, et al. Acute pancreatitis and organ failure:pathophysiology natural history,and management strategies[J].Curr Gastroenterol Rep, 2004,6(2):99-103
[18] Wittel UA, Rau B, Gansauge F, et al. Influence of PMN Leukocyte-Mediated Pancreatic Damage on the Systemic Immune Response in Severe Acute Pancreatitis in Rats. Dig Dis Sci [J]. 2004,49(8):1348-1357
[19] 傅强 , 崔乃强 , 邵伟 , 等. 急性出血坏死性胰腺炎小鼠辅助性 T 细胞亚群1/2 的 变 化 规 律 及 中 药 干 预 作 用 研 究 [J]. 中 国 中 西 医 结 合 急 救 杂 志2006,13(4):214-217
[20] Ma T, Kang CS, Shao HW, et al.Protective Effects of Ulinastatin on Proliferation and Cytokine Release of Splenocytes from Rats with Severe Acute Pancreatitis[J]. Eur Surg Res 2006,38(5):445-450
[21] 郑长青,胡刚正.CD4+T 细胞亚群的新认识及对炎症性肠病研究的指导[J].世界华人消化杂志,2004,12(3):505-5117
[22] Richard AG. Immunology[M]. 5th ed. New York:Freeman and Company, 2003: 288-291
[23] 安云庆,高晓明. 医学免疫学[M].北京:北京大学医学出版社,2004:74-78
[24] Takashi U, Yoshifumi T, Takeo Y, et al. Significant elevation of serum interleukin-18 levels in patients with acute pancreatitis[J]. J Gastroenterol,2006, 41(2):158-165
[25] Raffaele P, Rosa C, Bhajat B, et al. Immune-Manipulation of the Inflammatory Response in Acute Pancreatitis. What Can Be Expected? [J]. J Pancreas (Online) 2004,5(3):115-121
[26] Horwitz MS, Krahl T, Fine C, et al. Protection from Lethal Coxsackievirus- Induced Pancreatitis by Expression of Gamma Interferon[J]. J Virol, 1999 ,73(3): 1756-1766
[27] Rau BM, Kruger CM, Hasel C, et al. Effects of Immunosuppressive and Immunostimulative Treatment on Pancreatic Injury and Mortality in Severe Acute Experimental Pancreatitis[J]. Pancreas, 2006 ,33(2): 174-183
[28] Anne D, Jacques D .New Frontiers in the Pharmacological Prevention of Post-ERCP Pancreatitis: The Cytokines [J]. JOP, 2003 ,4(1):49-57.
[29] Zhang C, Ge CL, Guo RX, et al. Effect of IL-4 on altered expression of complement activation regulators in rat pancreatic cells during severe acute pancreatitis[J]. World J Gastroenterol. 2005, 11(43): 6770-6774
[30] Brock P , Gisela S, Thomas R, et al. Interleukin-4 gene transfer into rat pancreas byrecombinant adenovirus[J] . Scand J Gastroenterol, 2005,40(9): 1109-1117
[31] Raquel L, Juan M, Carlos M, et al.Different profile of cytokine synthesis according to the severity of acute pancreatitis[J]. World J Gastroenterol 2005,11(34): 5309-5313.
[32] Chao KC,Chao KF, Chuang CC, et al.Blockade of interleukin 6 accelerates acinar cell apoptosis and attenuates experimental acute pancreatitis in vivo[J]. Br J Surg, 2006,93(3):332-338
[33] Rarcity MG, Connor S, Criddle CN, et al. Acute pancreatitis and organ failure: panthophysiology natural history,and management strategies [J]. Curr Gastroenterol Rep, 2004,6(2):99-103
[34] Nora E. Cruz R, Carmen M B, et al. Toll-like receptors: dysregulation in vivo in patients with acute respiratory distress syndrome[J].Rev Alerg Mex,2004,51(6): 210-217
[35] Mentula PM. Kyl?np?? LE.Kemppainen S. et al.Plasma antiinflammatory cytokines and monocyte human leucocyte antigen-DR expression in patients with acute pancreatitis[J]. Scand J Gastroenterol,2004,39(2): 178-187
[36] Uwe AW,Bettina R,Frank G,et al.Influence of PMN Leukocyte-Mediated Pancrearic Damage on the Systemic Immune Response in Severe Acute Pancreatitis in Rats[J]. Dig Dis Sci, 2004,49(8):1348-1357
[37] Kylanpaa ML, Mentula P, Kemppainen E, et al.Monocyte Anergy Is Present in Patients With Severe Acute Pancreatitis and Is Significantly Alleviated by Granulocyte-Macrophage Colony-stimulating Factor and Interferon-[gamma] In Vitro[J]. Pancreas, 2005,31(1):23-27
[38] 江晓东,陈惠晓.Th 细胞的分化调控[J]. 国外医学·免疫学分册,2003,26(2):79-82
[39] Kristine R, Linda VD, Dale R, et al. Peptide YY Attenuates STAT1 and STAT3 Activation Induced by TNF-α in Acinar Cell Line AR42J[J]. J Am Coll Surg, 2006,202(5):788-796
[40] Ouyang W,Lohning M, Gao Z, et al.Stat6-independent GATA-3 autoactivation dirests IL-4-independent Th2 development and commitment[J]. Immunity, 2000, 12(1):27-37
[41] Dorfman DM, van den Elzen P, Weng AP, et al. Differential expression of T-bet,a T-box transcription factor required for Th1 T-cell development ,in perihheral T-cell lymphomas[J]. Am J Clin Pathol, 2003,120(6): 866-873
[2] Bonagura VR, Hatam L, DeVoti J, et a1. Recurrent respiratory papillom- atosis:altered CD8+ T-cell subsets and TH1/TH2 cytokine imba1ance[J].Clin Immunol,1999,93(3):302-3l1
[3] 姚新生.Thl 与 Th2 研究概况及进展[J].国外医学·免疫学分册,2002,25(6): 290-293
[4] 周家华,朱海涛,王凤臣,等.急性坏死性胰腺炎大鼠胰腺组织 NF-κB 活性变化及对其干预研究[J].东南大学学报:医学版,2007, 26(1):14-18
[5] Takahito H, Yuko I, Akihiko K, et al.IFN-gamma protects cerulein-induced acute pancreatitis by repressing NF-kappa B activation[J].J Immunol, 2007, 178(11): 7385-7394
[6] Bhatia M, Brady M, Shokubi S, et al. Inflammatory mediators in acute pancreatitis[J]. J Pathol, 2000,190(2):117-125
[7] Laser HG, Gross V, Scheibenbogen C, et al. Elevation of serum interleukin-6 concentration precedes acute phase response and reflects severity in acute pancreatitis [J] .Gastroenterology, 1991,101:782-785
[8] 安云庆,高晓明.医学免疫学[M].北京:北京大学医学出版社,2004:74-78
[9] Wrobleski DM, Barth MM,Oyen LJ .Necrotizing pancreatitis pathophysi- ology,diagnosis, and acute care management[J].AACN Clin Issues, 1999,10(4): 464-477.
[10] 吕云福.现代胰腺外科学[M].北京:人民军医出版社,2003:126-134
[11] Ueda T, Takeyama Y, Yasuda T, et al. Functional alterations of splenocytes in severe acute pancreatitis [J]. J Surg Res, 2002, 102(2) :161-168
[12] Ueda T ,Takeyama Y, Yasuda T, et al. Significant elevation of serum interleukin-18 levels in patients with acute pancreatitis[J].J Gastroenterol, 2006,41(2):158-165
[13] Richard AG. Immunology[M]. 5th ed. New York:Freeman and Company, 2003:288-291
[14] 江晓东,陈惠晓.Th 细胞的分化调控[J].国外医学·免疫学分册,2003,26(2): 79-82
[15] Ouyang W,Lohning M, Gao Z, et al. Stat6-Independent GATA-3 Autoactivation Directs IL-4-Independent Th2 Development and Commitment [J]. Immunity, 2000,12(1):27-37
[16] Dorfman DM, van den Elzen P, Weng AP, et al. Differential expression of T-bet,a T-box transcription factor required for Th1 T-cell development ,in perihheral T-cell lymphomas[J]. Am J Clin Pathol, 2003, 120(6): 866-873
[17] Rarcity MG,Connor S,Criddle CN, et al. Acute pancreatitis and organ failure:pathophysiology natural history,and management strategies[J].Curr Gastroenterol Rep, 2004,6(2):99-103
[18] Wittel UA, Rau B, Gansauge F, et al. Influence of PMN Leukocyte-Mediated Pancreatic Damage on the Systemic Immune Response in Severe Acute Pancreatitis in Rats. Dig Dis Sci [J]. 2004,49(8):1348-1357
[19] 傅强 , 崔乃强 , 邵伟 , 等. 急性出血坏死性胰腺炎小鼠辅助性 T 细胞亚群1/2 的 变 化 规 律 及 中 药 干 预 作 用 研 究 [J]. 中 国 中 西 医 结 合 急 救 杂 志2006,13(4):214-217
[20] Ma T, Kang CS, Shao HW, et al.Protective Effects of Ulinastatin on Proliferation and Cytokine Release of Splenocytes from Rats with Severe Acute Pancreatitis[J]. Eur Surg Res 2006,38(5):445-450
[21] 郑长青,胡刚正.CD4+T 细胞亚群的新认识及对炎症性肠病研究的指导[J].世界华人消化杂志,2004,12(3):505-5117
[22] Richard AG. Immunology[M]. 5th ed. New York:Freeman and Company, 2003: 288-291
[23] 安云庆,高晓明. 医学免疫学[M].北京:北京大学医学出版社,2004:74-78
[24] Takashi U, Yoshifumi T, Takeo Y, et al. Significant elevation of serum interleukin-18 levels in patients with acute pancreatitis[J]. J Gastroenterol,2006, 41(2):158-165
[25] Raffaele P, Rosa C, Bhajat B, et al. Immune-Manipulation of the Inflammatory Response in Acute Pancreatitis. What Can Be Expected? [J]. J Pancreas (Online) 2004,5(3):115-121
[26] Horwitz MS, Krahl T, Fine C, et al. Protection from Lethal Coxsackievirus- Induced Pancreatitis by Expression of Gamma Interferon[J]. J Virol, 1999 ,73(3): 1756-1766
[27] Rau BM, Kruger CM, Hasel C, et al. Effects of Immunosuppressive and Immunostimulative Treatment on Pancreatic Injury and Mortality in Severe Acute Experimental Pancreatitis[J]. Pancreas, 2006 ,33(2): 174-183
[28] Anne D, Jacques D .New Frontiers in the Pharmacological Prevention of Post-ERCP Pancreatitis: The Cytokines [J]. JOP, 2003 ,4(1):49-57.
[29] Zhang C, Ge CL, Guo RX, et al. Effect of IL-4 on altered expression of complement activation regulators in rat pancreatic cells during severe acute pancreatitis[J]. World J Gastroenterol. 2005, 11(43): 6770-6774
[30] Brock P , Gisela S, Thomas R, et al. Interleukin-4 gene transfer into rat pancreas byrecombinant adenovirus[J] . Scand J Gastroenterol, 2005,40(9): 1109-1117
[31] Raquel L, Juan M, Carlos M, et al.Different profile of cytokine synthesis according to the severity of acute pancreatitis[J]. World J Gastroenterol 2005,11(34): 5309-5313.
[32] Chao KC,Chao KF, Chuang CC, et al.Blockade of interleukin 6 accelerates acinar cell apoptosis and attenuates experimental acute pancreatitis in vivo[J]. Br J Surg, 2006,93(3):332-338
[33] Rarcity MG, Connor S, Criddle CN, et al. Acute pancreatitis and organ failure: panthophysiology natural history,and management strategies [J]. Curr Gastroenterol Rep, 2004,6(2):99-103
[34] Nora E. Cruz R, Carmen M B, et al. Toll-like receptors: dysregulation in vivo in patients with acute respiratory distress syndrome[J].Rev Alerg Mex,2004,51(6): 210-217
[35] Mentula PM. Kyl?np?? LE.Kemppainen S. et al.Plasma antiinflammatory cytokines and monocyte human leucocyte antigen-DR expression in patients with acute pancreatitis[J]. Scand J Gastroenterol,2004,39(2): 178-187
[36] Uwe AW,Bettina R,Frank G,et al.Influence of PMN Leukocyte-Mediated Pancrearic Damage on the Systemic Immune Response in Severe Acute Pancreatitis in Rats[J]. Dig Dis Sci, 2004,49(8):1348-1357
[37] Kylanpaa ML, Mentula P, Kemppainen E, et al.Monocyte Anergy Is Present in Patients With Severe Acute Pancreatitis and Is Significantly Alleviated by Granulocyte-Macrophage Colony-stimulating Factor and Interferon-[gamma] In Vitro[J]. Pancreas, 2005,31(1):23-27
[38] 江晓东,陈惠晓.Th 细胞的分化调控[J]. 国外医学·免疫学分册,2003,26(2):79-82
[39] Kristine R, Linda VD, Dale R, et al. Peptide YY Attenuates STAT1 and STAT3 Activation Induced by TNF-α in Acinar Cell Line AR42J[J]. J Am Coll Surg, 2006,202(5):788-796
[40] Ouyang W,Lohning M, Gao Z, et al.Stat6-independent GATA-3 autoactivation dirests IL-4-independent Th2 development and commitment[J]. Immunity, 2000, 12(1):27-37
[41] Dorfman DM, van den Elzen P, Weng AP, et al. Differential expression of T-bet,a T-box transcription factor required for Th1 T-cell development ,in perihheral T-cell lymphomas[J]. Am J Clin Pathol, 2003,120(6): 866-873