Effect of IL-4 on altered expression of complement activation regulators in rat pancreatic cells during severe acute pancreatitis

AIM: To investigate the effect of IL-4 on the altered expression of complement activation regulators in pancreas and pancreatic necrosis during experimental severe acute pancreatitis (SAP). METHODS: SAP model of rats was established by retrograde injection of 5% sodium taurocholate (1 mL/kg) into the pancreatic duct. We immunohistochemically assayed the expression of three complement activation regulators: decay accelerating factor (DAF; CD55), 20 ku homologous restriction factor (HRF20; CD59) and membrane cofactor protein (MCP; CD46), in the pancreatic acinar cells of rats at 0, 3, 6, 12, and 24 h after the induction of SAP model. Meanwhile the levels of amylase and lipase were determined, and morphological examination was performed. Then, 61 rats were randomly divided into three groups. Group A (n = 21) received no treatment after the SAP model was established; group B (n = 20) was given IL-4 (8 μg/animal) intraperitoneally 0,5 h before the SAP model was established; group C (n = 20) was given IL-4 (8 μg/animal) intraperitoneally 0.5 h after the SAP model was established. Plasma amylase and lipase, extent of pancreatic necrosis and expression of complement activation regulators were investigated 6 h after the induction of SAP model. RESULTS: Three complement activation regulators were all expressed in pancreatic acinar cells. MCP was not found on the basolateral surface as reported. Contrary to the gradually increasing plasma level of amylase and lipase, expression of complement activation regulators decreased after SAP model was set up. At the same time, the severity of pancreatic necrosis was enhanced. A strong negative correlation was found between the ex- pression of MCP, DAF, CD59 in pancreatic acinar cells and the severity of pancreatic necrosis (r=-0.748, -0.827, -0.723; P＜0.01). In the second series of experiments, no matter when the treatment of IL-4 was given (before or after the induction of SAP model), the serum level of amylase or lipase was decreased and the extent of pancreatic necrosis was ameliorated significantly. Compared to SAP control group, the expression of DAF and CD59 in pancreas was reinforced when IL-4 was given before the induction of SAP model (P＜0.01, P＜0.05), but the expression of MCP was not influenced (P＞0.05). The expression of DAF was enhanced, when IL-4 was given after the induction of SAP model (P＜0.05), but the expression of CD59 and MCP did not change (P＞0.05). CONCLUSION: Complement activation regulators may participate in the pathogenesis of pancreatic inflammation. Downregulation of complement activation regulators expression may be one of the causes of pancreatic necrosis. IL-4 treatment may control SAP aggravation by enhancing expression of DAF and CD59 in pancreas and decreasing pancreatic necrosis. Moreover, DAF and CD59 may play an important role in the regulation of complement activation regulators during SAP.     

Leptin treatment ameliorates acute lung injury in rats with cerulein-induced acute pancreatitis

AIM:To determine the effect of exogenous leptin on acute lung injury (ALI) in cerulein-induced acute pancreatitis (AP). METHODS:Forty-eight rats were randomly divided into 3 groups. AP was induced by intraperitoneal (i.p.) injection of cerulein (50 μg/kg) four times,at 1 h intervals. The rats received a single i.p. injection of 10 μg/kg leptin (leptin group) or 2 mL saline (AP group) after cerulein injections. In the sham group,animals were given a single i.p. injection of 2 mL saline. Experimental samples were collected for biochemical and histological evaluations at 24 h and 48 h after the induction of AP or saline administration. Blood samples were obtained for the determination of amylase,lipase,tumor necrosis factor (TNF)-a,interleukin (IL)-1β,macrophage inflammatory peptide (MIP)-2 and soluble intercellular adhesion molecule (sICAM)-1 levels,while pancreatic and lung tissues were removed for myeloperoxidase (MPO) activity,nitric oxide (NOx) level,CD40 expression and histological evaluation. RESULTS:Cerulein injection caused severe AP,confirmed by an increase in serum amylase and lipase levels,histopathological findings of severe AP,and pancreatic MPO activity,compared to the values obtained in the sham group. In the leptin group,serum levels of MIP-2,sICMA-1,TNF-a,and IL-1b,pancreatic MPO activity,CD40 expression in pancreas and lung tissues,and NOx level in the lung tissue were lower compared to those in the AP group. Histologically,pancreatic and lungdamage was less severe following leptin administration. CONCLUSION:Exogenous leptin attenuates inflammatory changes,and reduces pro-inflammatory cytokines,nitric oxide levels,and CD40 expression in ceruleininduced AP and may be protective in AP associated ALI.     

抗氧化剂对大鼠重症急性胰腺炎胰腺腺泡细胞凋亡的影响

目的探讨四氢化吡咯二硫代氨基甲酸酯（PDTC）对重症急性胰腺炎（SAP）胰腺腺泡细胞凋亡的影响。方法SD大鼠72只，随机分为：假手术组（sham operation，SO）、SAP组和PDTC组。SAP模型采用5％牛磺胆酸钠1ml/kg胰胆管逆行穿刺注射建立，PDTC组在造模前1h给予腹腔注射PDTC（1130mg/kg），术后分4个时段（1、3、6、12h）分批进行腹主动脉采血后处死，取胰腺组织作病理切片与液氮冻存。胰腺腺泡细胞的凋亡检测应用TUNEL法、电镜以及免疫组化法检测胰腺组织Caspase-3的表达；核因子（NF）-κB活化的检测应用免疫组化法。同时观察各组血清淀粉酶、脂肪酶水平及胰腺组织病理学评分。结果SAP组各时间点血清淀粉酶及脂肪酶水平及胰腺组织病理组织学评分较SO组显著增高（P〈0．05）。PDTC治疗组血清淀粉酶、脂肪酶水平较SAP组明显降低；胰腺组织病理组织学评分明显改善。PDTC治疗后3、6、12h胰腺组织Caspase-3的表达显著高于SAP组（P〈0．01），而在1h无统计学差异；各时间点胰腺腺泡细胞凋广指数显著高于SAP组（P〈0．01）；NF—κB的活化显著低于SAP组（P〈0．01）。结论SAP时，PDTC可能通过抑制NF—κB的活化，从而抑制NF—κB介导胰腺细胞的抗凋亡效应，减少胰腺腺泡细胞坏死。     

中药鸦胆子与补骨脂素对卡氏肺孢子虫病大鼠免疫调节的影响

目的 探讨中药鸦胆子和补骨脂素合剂对卡氏肺孢子虫肺炎(PCP)大鼠的免疫调节及杀虫作用。方法 地塞米松皮下注射SD大鼠(2.5mg/只,每周2次,连续6周),建立PCP大鼠模型。药物治疗剂量,每鼠每天口饲中药鸦胆子0.12mg与补骨脂素1mg,连续7d或14d。同时设PCP阳性不治疗对照组及正常大鼠对照组。制作肺组织病理切片观察肺组织病变,制作肺组织印片计数肺孢子虫包囊数观察中药合剂杀虫效果。检测血液中CD4⁺T细胞、CD8+T细胞和α肿瘤坏死因子(TNF-α)的变化,观察中药合剂对PCP大鼠的免疫调节作用。结果 治疗组大鼠,受损的肺组织病变减轻或修复,体重明显回升,肺组织包囊数明显低于阳性对照组(P<0.05)。血液中CD4⁺T细胞、CD8⁺T细胞和TNF-α均较阳性对照组升高(P<0.05)。结论 中药鸦胆子和补骨脂素合剂治疗PCP大鼠,可增强免疫调节作用,并可抑制及杀灭卡氏肺孢子虫。     

Can albumin administration relieve lung injury in trauma/hemorrhagic shock？

INTRODUCTION Previous studies have shown that there is obvious pulmonary microvascular injury at the early stage of trauma/hemorrhagic shock (T/HS)[1]. The polymorpho- nuclear neutrophils (PMNs) accumulated in lung are closely correlated with lung injury[…     

支气管哮喘大鼠CD4+CD25+T细胞的变化及地塞米松干预作用的研究

目的 研究支气管哮喘(简称哮喘)大鼠模型支气管肺泡灌洗液(BALF)、血液、脾脏CD4+CD25+T细胞的变化,及地塞米松对CD4+CD25+T细胞的影响.方法 50只SD大鼠随机分为5组,空白对照(A)组,哮喘(B)组,地塞米松1(C)组、地塞米松2(D)组,地塞米松3(E)组.A组第l天给予腹腔注射生理盐水l ml,第15～21天每天给予生理盐水雾化.B、C、D、E组用卵蛋白建立哮喘大鼠模型,第1天,每只大鼠腹腔注射抗原l ml(卵蛋白1 mg+灭活百日咳杆菌9×106个+氢氧化铝干粉100 mg)混悬液,第15～21天给予1%的卵蛋白雾化30 min,C、D、E组于雾化后分别给予腹腔注射地塞米松0.2 mg/kg、1 mg/kg、2 mg/kg.采用流式细胞仪检测的方法 ,观察大鼠体内BALF、外周血、脾脏CD4+CD25+T细胞的变化及使用不同剂量地塞米松后对其的影响.结果 B组BALF、外周血、脾脏CD4+CD25+T细胞表达占CD4+T细胞的百分比分别是(42.21±5.62)%、(12.69±2.70)%、(11.15±1.05)%,A组结果 分别是(18.76±5.85)%、(6.21±1.73)%、(7.85±2.13)%.B组与A组比较,差异均具有统计学意义(P＜0.01,P＜0.01,P＜0.05);C组、D组、E组BALF中CD4+CD25+T细胞占CD4+T细胞的百分比表达分别是(10.49±4.03)%、(13.28±5.12)%、(7.51±5.39)%,显著低于A组和B组,(P＜0.05,P＜0.01);外周血中,C组(6.03±1.43)%、D组(4.88±0.95)%与A组(6.21±1.73)%比较,差异无统计学意义,E组(3.49±0.62)%与C组、A组比较,差异有统计学意义(P＜0.05).脾脏中,C组(7.25±1.82)%、D组(8.63±3.18)%与A组(7.85±2.13)%比较,差异无统计学意义,E组(3.38±1.37)%与C组、D组、A组比较,差异有统计学意义(P＜0.05).结论 CD4+CD25+T细胞在哮喘大鼠体内有明显的优势表达,可能是哮喘发病的机制之一.地塞米松可以抑制CD4+CD25+T细胞的表达.BALF内CD4+CD25+T细胞的变化与外周血和脾脏的变化具有一致性,监测外周血或脾脏CD4+CD25+T细胞变化可了解肺部情况.     

支气管哮喘大鼠CD4^＋CD25^＋T细胞的变化及地塞米松干预作用的研究

目的研究支气管哮喘（简称哮喘）大鼠模型支气管肺泡灌洗液（BALF）、血液、脾脏CD4^＋CD25^＋T细胞的变化,及地塞米松对CD4^＋CD25^＋T细胞的影响。方法50只SD大鼠随机分为5组,空白对照（A）组,哮喘（B）组,地塞米松1（C）组、地塞米松2（D）组,地塞米松3（E）组。A组第1天给予腹腔注射生理盐水1ml,第15～21天每天给予生理盐水雾化。B、C、D、E组用卵蛋白建立哮喘大鼠模型,第1天,每只大鼠腹腔注射抗原1ml（卵蛋白1mg＋灭活百日咳杆菌9×10。个＋氢氧化铝干粉100mg）混悬液,第15～21天给予1％的卵蛋白雾化30min,C、D、E组于雾化后分别给予腹腔注射地塞米松0．2mg／kg、1mg／kg、2mg／kg。采用流式细胞仪检测的方法,观察大鼠体内BALF、外周血、脾脏CD4^＋CD25^＋T细胞的变化及使用不同剂量地塞米松后对其的影响。结果B组BALF、外周血、脾脏CD4^＋CD25^＋T细胞表达占CD4^＋T细胞的百分比分别是（42．21±5．62）％、（12．69±2．70）％、（11．15±1．05）％,A组结果分别是（18．76±5．85）％、（6．21±1．73）％、（7．85±2．13）％。B组与A组比较,差异均具有统计学意义（P〈0．01,P〈0．01,P〈0．05）;C组、D组、E组BALF中CD4^＋CD25^＋T细胞占CD4^＋T细胞的百分比表达分别是（10．49±4．03）oA、（13．28±5．12）％、（7．51±5．39）％,显著低于A组和B组,（P〈0．05,P〈0．01）;外周血中,C组（6．03±1．43）％、D组（4．88±0．95）％与A组（6．21±1．73）％比较,差异无统计学意义,E组（3．49士0．62）％与C组、A组比较,差异有统计学意义（P〈0．05）。脾脏中,c组（7．25±1．82）%、D组（8．63±3．18）％与A组（7．85±2．13）％比较,差异无统计学意义,E组（3．38±1．37）％与C组、D组、A组比较,差异有统计学意义（P〈0．05）。结论CD4^＋CD25^＋T细胞在哮喘大鼠体内有明显的优势表达,可能是哮喘发病的机制之一。地塞米松可以抑制CD4^＋CD25^＋T细胞的表达。BALF内CD4^＋CD25^＋T细胞的变化与外周血和脾脏的变化具有一致性,监测外周血或脾脏CD4^＋CD25^＋T细胞变化可了解肺部情况。     

T细胞亚群在TNBS诱发大鼠实验性结肠炎发病及治疗后的变化

目的探讨T细胞亚群在TNBS诱发大鼠实验性结肠炎模型的外周血、脾、结肠中变化特点及经柳氮磺胺吡啶（SASP）和氢化泼尼松（PSL）治疗后这些细胞的变化。方法分为对照组、建模后1周及3周组、SASP及PSL治疗组，用流式细胞仪检测各组外周血、脾和结肠黏膜上皮细胞内CD4^＋，CD8^＋T细胞比例的变化；肠道积分评定组织学变化。结果CD4^＋T细胞：建模第1周外周血、脾明显低于对照组；建模第3周脾CD4^＋T细胞升至对照组水平，SASP治疗组外周血高于模型3周组，PSL治疗组外周血、脾、结肠均明显低于对照组和模型3周组。CD8^＋T细胞：建模第1周外周血、结肠低于对照组，建模第3周外周血中仍低，结肠升至对照组水平；SASP治疗组外周血高于模型3周组。PSL治疗组外周血、脾、结肠均明显低于对照组，脾、结肠低于模型3周组。结论大鼠实验性结肠炎发病过程中，机体免疫功能下调，以局部免疫异常为主，与整体的免疫异常可能关系较小。PSL导致机体免疫功能下调。SASP对全身免疫状态影响较小，其作用更集中于肠道。     

弓形虫感染大鼠外周血T淋巴细胞亚群及IFN-γ、TNF-α、IL-4动态变化

目的 观察弓形虫感染大鼠外周血T淋巴细胞亚群及干扰素-γ（IFN-γ）、肿瘤坏死因子-α（TNF-α）、白细胞介素-4（IL-4）动态变化。 方法 将54只清洁级Sprague-Dawley（SD）成年大鼠随机分成9组,其中8组每鼠腹腔注射经纤维素-11（CF-11）纯化的活弓形虫速殖子悬液2 ml（2×10⁵/L）。分别于弓形虫感染后1、3、7、14、28、35、42、60 d时取大鼠外周全血,流式细胞术检测T细胞亚群变化,ELISA检测外周血IFN-γ、TNF-α、IL-4动态变化。对照组腹腔注射2 ml生理盐水。 结果 T淋巴细胞亚群CD8+水平在感染弓形虫后3 d（14.22%）、7 d（14.93%）、14 d（16.08%）均显著低于正常水平（23.08%）（P＜0.05）,至感染后28 d恢复正常水平,而CD4+水平在整个感染过程中无明显变化。IFN-γ和IL-4水平在感染后第7 d分别升高至6.73 pg/ml和6.91 pg/ml（P＜0.05）,至感染60 d仍高于正常水平;TNF-α至感染后28 d（14.37 pg/ml）及60 d均高于正常水平（10.81 pg/ml）（P＜0.05）。 结论 弓形虫感染后大鼠外周血CD8+水平一度降低,28 d后恢复正常,CD4+无明显变化,IFN-γ、IL-4和TNF-α均高于正常水平,但出现时间有差异。     

Effect of resveratrol on activation of nuclear factor kappa-B and inflammatory factors in rat model of acute pancreatitis

AIM: To observe the effect of resveratrol on nuclear factor Kappa-B (NF-κB) activation and the inflammatory response in sodium taurocholate-induced pancreatitis in rats. METHODS: Seventy-two male SD rats were randomly divided into three groups: sham operation group (control), severe acute pancreatitis (SAP) group, and severe acute pancreatitis group treated with resveratrol (RES). A SAP model was established by injecting 4% sodium taurocholate 1 mL/kg through puncturing the pancreatic duct. In Res group, Res was given at 30 mg/kg b.m. intraperitoneally after the SAP model was successfully established. Eight animals from each group were sacrificed at 3, 6 and 12 h after modeling. The expression of NF-κB activation of pancreas was detected by irnmunohistochemical staining, whereas the levels of TNF-α and IL-8 in pancreatic tissues were estimated by radioimrnunoassay. The pathological changes of pancreas and lungs were examined microscopically. RESULTS: Much less hyperemia, edema, dust-colored necrotic focus and soaps were noticed in pancreas in RES group than in SAP group. In RES group, hemorrhage, exudates and infiltration of inflammatory cells in pancreas and interstitial edema, destruction of alveolar wall in lung were significantly less than in SAP group. In the SAP group, the activation of NF-κB in pancreatic tissues was enhanced significantly at any measure point compared with control group (64.23±10.72% vs 2.56±0.65%, 55.86±11.34% vs 2.32±0.42%, 36.23±2.30% vs 2.40±0.36%,P<0.01), TNF-α, IL-8 were also increased and reached their peak at 6 h and then declined. The activation of NF-κB and the levels of TNF-α and IL-8 in RES group were significantly lower than those in SAP group (P<0.01): activation (52.63±9.45% vs 64.23±10.72%, 40.52±8.40% vs 55.86±11.34%, 29.83±5.37% vs 36.23±2.30%), TNF-α (132.76±15.68 pg/mL vs 158.36±12.58 pg/mL, 220.32±23.57 pg/mL vs 247.67± 11.62 pg/mL, 175.68±18.43 pg/mL vs 197.35±12.57 pg/mL) and IL-8 (0.62±0.21 μg/L vs 0.83±0.10 μg/L, 1.10±0.124 μg/L vs1.32±0.18 μg/L, 0.98±0.16 μg/L vs 1.27±0.23μg/L). CONCLUSION: The activation of NF-KB is involved in the inflammatory response of rats with SAP. Resveratrol could effectively inhibit the expression of NF-κB activation, alleviate the severity of SAP through its anti-inflammatory effects and regulate the inflammatory mediators.     

Activated protein C,an anticoagulant polypeptide,ameliorates severe acute pancreatitis via regulation of mitogen-activated protein kinases

Background Our aim was to investigate the changes of mitogen-activated protein kinases (MAPKs) by activated protein C (APC) treatment in rats with severe acute pancreatitis (SAP), and relate them to changes in SAP severity, thus providing evidence for developing clinical therapies. Methods Sprague-Dawley rats were given an intravenous injection of saline (SAP group), APC (50 μg/kg or 10 μg/kg), or CNI1493 just before SAP induction. One group of rats underwent a sham operation (control group). Experimental samples were harvested 16 h after SAP induction. The gene expression of pancreatic MAPKs was evaluated by cDNA microarrays. The mRNA and protein/phosphorylated protein levels of p38 MAPK, extracellular signal-regulated protein kinase (ERK) 1/2, and c-Jun N-terminal kinase (JNK) and the protein levels of tumor necrosis factor (TNF)-α and interleukin (IL)-1β were determined in pancreatic tissue. The severity of disease was evaluated by pancreatic histology, the pancreatic wet/dry weight ratio, and the serum amylase level. Results In rats treated with APC (50 μg/kg) or CNI1493, the severity of pancreatitis and expression of pancreatic TNF-α and IL-1β proteins were attenuated by the decreased expression and activity of p38 MAPK and JNK (vs. the SAP group, P < 0.01). The expression and activity of ERK1/2 were increased in APC-treated rats, especially in the group treated with APC 50 μg/kg (vs. the SAP or CNI1493-treated group, P < 0.01, respectively). Conclusions Inhibition of expression of pancreatic p38 MAPK and JNK and upregulation of ERK1/2 expression by APC treatment may protect against pancreatic injury, thus ameliorating severity of the disease.     

艾滋病患者胃黏膜与外周血中人类免疫缺陷病毒感染和CD4~+T淋巴细胞数量的区室性差异的探讨

目的 探讨AIDS患者胃黏膜与外周血中HIV感染和CD4~+T淋巴细胞数量的区室性差异.方法 选取AIDS患者35例,对照组为HIV抗体阴性者10例,均进行胃镜检查并收集外周血.PCR法制备地高辛标记HIV-1长末端重复序列(LTR)、gag基因的双链cDNA探针,核酸原位杂交方法观察胃黏膜组织冰冻切片和外周血单个核细胞(PBMC)涂片H1V感染情况,免疫组织化学方法检测CD41T淋巴细胞,数据结果行t检验.结果 AIDS未治疗组胃黏膜中HIV阳性率为(1.67±1.48)%,PBMC中为(19.37±9.23)%.AIDS未治疗组与高效抗反转录病毒治疗(HAART)组各组间的胃黏膜HIV阳性率差异尤统计学意义(t=-0.996,t=-0.794,t=-0.461;P＞0.05).PBMC涂片中,治疗1～4年组HIV阳性率为(4.25±3.47)%,明显低于未治疗组的(19.37±9.23)%(t=3.000,P＜0.05).AIDS未治疗组胃黏膜单个核细胞(MMC)中CD4+T淋巴细胞阳性率为(12.53±8.14)%,PBMC中CD4+T淋巴细胞阳性率为(19.00±9.55)%,HAART1～4年组胃黏膜MMC中CD4~+T淋巴细胞计数为(37.44±18.00)%,仍低于对照组的(50.35±3.41)%(t=-4.620,P＜0.01),但PBMC中CD4+T淋巴细胞计数与对照组比较,差异无统计学意义(t=-2.094,P＞0.05).结论 胃黏膜与外周血中HIV感染和CD4~+T淋巴细胞数量存在区室性差异.     

Alterations of intestinal immune function and regulatory effects of L-arginine in experimental severe acute pancreatitis rats

ABM: To discuss the changes of intestinal mucosal immune function in rats with experimental severe acute pancreatitis (SAP) and the regulatory effect of L-arginine. METHODS: Male adult Wistar rats were randomly divided into pancreatitis group, sham-operation group, and L-arginine treatment group. Animals were killed at 24, 48, and 72 h after SAP models were developed and specimens were harvested. Endotoxin concentration in portal vein was determined by limulus endotoxin analysis kit. CD3+, CD4+, CD8+ T lymphocytes in intestinal mucosal lamina propria were examined by immunohistochemistry. Secretory immunoglobulin A (SIgA) in cecum feces was examined by radioimmunoassay. RESULTS: Compared to the control group, plasma endotoxin concentration in the portal vein increased, percentage of CD3+ and CD4+ T lymphocyte subsets in the end of intestinal mucosal lamina propria reduced significantly, CD4+/CD8+ ratio decreased, and SIgA concentrations in cecum feces reduced at 24, 48, and 72 h after SAP developed. Compared to SAP group, the L-arginine treatment group had a lower level of plasma endotoxin concentration in the portal vein, a higher CD3+ and CD4+ T lymphocyte percentage in the end of intestinal mucosal lamina propria, an increased ratio of CD4+/CD8+ and a higher SIgA concentration in cecum feces. CONCLUSION: Intestinal immune suppression occurs in the early stage of SAP rats, which may be the main reason for bacterial and endotoxin translocation. L-arginine can improve the intestinal immunity and reduce bacterial and endotoxin translocation in SAP rats.     

大鼠卡氏肺孢子虫肺炎的免疫应答

[目的 ]探讨应用糖皮质激素 (GC)诱发大鼠卡氏肺孢子虫肺炎 (Pneumocystiscariniipneumonia ,PCP)的免疫应答。 [方法 ]SD大鼠皮下注射GC建立免疫抑制PCP模型 ,测定外周血淋巴细胞比例、CD4+ T细胞 /CD8+ T细胞比例 ,肺泡灌洗液 (bronchoalveolarlavagefluid ,BALF)中淋巴细胞比例、sIL 2R和TNF α值。 [结果 ]①免疫抑制后外周血淋巴细胞比例、CD4+ T细胞 /CD8+ T细胞比例降低 ,BALF中淋巴细胞比例、TNF α、sIL 2R均降低 ;②BALF中TNF α、外周血CD4+ T细胞 /CD8+ T细胞比值在PCP组中最低 ;③PCP组的BALF中淋巴细胞比例比PC阴性对照组明显升高。 [结论 ]使用GC造成大鼠免疫抑制 ,诱发PC感染后 ,CD4+ 减少及TNF α分泌降低。     

Role of bone marrow-derived mesenchymal stem cells in a rat model of severe acute pancreatitis

AIM: To investigate the role and potential mechanisms of bone marrow mesenchymal stem cells (MSCs) in severe acute peritonitis (SAP).METHODS: Pancreatic acinar cells from Sprague Dawley rats were randomly divided into three groups: non-sodium deoxycholate (SDOC) group (non-SODC group), SDOC group, and a MSCs intervention group (i.e., a co-culture system of MSCs and pancreatic acinar cells + SDOC). The cell survival rate, the concentration of malonaldehyde (MDA), the density of superoxide dismutase (SOD), serum amylase (AMS) secretion rate and lactate dehydrogenase (LDH) leakage rate were detected at various time points. In a separate study, Sprague Dawley rats were randomly divided into either an SAP group or an SAP + MSCs group. Serum AMS, MDA and SOD, interleukin (IL)-6, IL-10, and tumor necrosis factor (TNF)-α levels, intestinal mucosa injury scores and proliferating cells of small intestinal mucosa were measured at various time points after injecting either MSCs or saline into rats. In both studies, the protective effect of MSCs was evaluated.RESULTS: In vitro, The cell survival rate of pancreatic acinar cells and the density of SOD were significantly reduced, and the concentration of MDA, AMS secretion rate and LDH leakage rate were significantly increased in the SDOC group compared with the MSCs intervention group and the Non-SDOC group at each time point. In vivo, Serum AMS, IL-6, TNF-α and MAD level in the SAP + MSCs group were lower than the SAP group; however serum IL-10 level was higher than the SAP group. Serum SOD level was higher than the SAP group at each time point, whereas a significant between-group difference in SOD level was only noted after 24 h. Intestinal mucosa injury scores was significantly reduced and the proliferating cells of small intestinal mucosa became obvious after injecting MSCs.CONCLUSION: MSCs can effectively relieve injury to pancreatic acinar cells and small intestinal epithelium, promote the proliferation of enteric epithelium and repair of the mucosa, attenuate systemic inflammation in rats with SAP.     

Interleukin-10-secreting CD4 cells from aged patients with AIDS decrease in-vitro HIV replication and tumour necrosis factor alpha production

OBJECTIVE: To evaluate the impact of age on the proliferative response, cytokine profile and viral kinetics in AIDS patients treated successfully with antiretroviral drugs. METHODS: Peripheral blood mononuclear cells (PBMC), CD4 cell-depleted PBMC or CD4 T cells from young adult and aged HIV-1-infected patients were activated in vitro with anti-CD3 with or without interleukin (IL)-2. Lymphoproliferation and cytokines were measured after 3 days and in-vitro HIV-1 replication after 7 days. RESULTS: Both lymphoproliferation and cytokine [IL-1beta, tumour necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma)] secretion were higher in younger than in older AIDS patients. In cultures of cells derived from aged patients and activated by anti-CD3, IFN-gamma production was severely damage and IL-10 production was much higher. Although IL-2 addition to activated PBMC elevated IFN-gamma secretion, IL-10 production remained elevated in the aged group. The depletion of CD4 T lymphocytes from these cultures dramatically reduced released IL-10 in the older group but did not alter significantly IFN-gamma production. Interestingly, higher IL-10 levels produced by CD4 T cells were related to lower in-vitro HIV-1 replication, and the blockade of this cytokine by anti-IL-10 monoclonal antibody enhanced virus replication. This effect may be correlated with elevated TNF-alpha secretion. Finally, impaired IFN-gamma secretion detected in activated CD4 T cells obtained from aged patients was not directly correlated with high IL-10 production. CONCLUSIONS: Elevated IL-10 production by aged AIDS patients contributed considerably to control of HIV replication and to inhibition of TNF-alpha secretion but not to the reduced IFN-gamma production.     

Effects of CH-100, a chinese herbal medicine, on acute concanavalin A-mediated hepatitis in control and alcohol-fed rats

BACKGROUND: Administration of concanavalin A (Con A) leads to acute hepatitis that involves T-cell activation and inflammatory mediator production in mice and rats. We examined the role of CH-100, a Chinese herbal medicine previously trialed in human hepatitis C, in the prevention of Con A-related, T-cell-mediated, acute liver injury in rats. METHODS: Female Wistar rats were fed 40% ethanol, 2% sucrose, or isocaloric sucrose for 8 weeks. At the same time, these animals were fed either the Chinese herbal medicine CH-100 (4 tablets/kg body weight/ day) or placebo in chow daily. Blood from the tail vein was collected for endotoxin (lipopolysaccharide) assay at 0, 4, and 8 weeks of ethanol consumption. Twenty-four hours after injection of Con A (20 mg/kg body weight) or phosphate-buffered saline, blood from the tail vein was collected for alanine aminotransferase and tumor necrosis factor (TNF)-alpha assays. Liver-associated CD4+ T cells were isolated from liver perfusates and then cultured with Con A (5 microg/ml) at 37 degrees C for 24 hr. Supernatants were harvested for TNF-alpha assay. The proportion of CD4+ T cells in blood and liver perfusates was measured. Liver samples were collected for histopathological analysis. RESULTS: Lipopolysaccharide levels were significantly reduced in CH-100-treated ethanol-fed rats compared with placebo-treated rats. After Con A injection, alanine aminotransferase levels were lower at 12 and 24 hr in herb-treated rats compared with placebo-treated rats. Furthermore, serum TNF-alpha levels were lower in ethanol-fed rats on herbal treatment. A significant decrease in TNF-alpha production by liver-associated CD4+ T cells in culture was observed in CH-100-treated ethanol-fed rats. CH-100 treatment was associated with a decreased percentage of CD4+ cells in both blood and liver perfusate in all groups. Herb-treated rats displayed markedly less hepatic necrosis and a reduced CD4+ T-cell infiltrate in portal areas than did placebo-fed rats. CONCLUSIONS: The results demonstrate that CH-100 modified the T-cell response to Con A injection. The effect was more marked in ethanol-fed rats, which suggests a possible role for CH-100 in treating alcoholic liver disease.     

外周血单个核细胞表面CD44表达水平与冠心病及其严重程度有关联

目的:观察不同类型冠心病患者外周血单个核细胞(PBMC)表面CD44的表达变化情况,并探讨其临床意义。方法:采用实时荧光定量RT-PCR法,对正常对照30例,稳定型心绞痛(SAP)26例,不稳定型心绞痛(UAP)29例、急性心肌梗死(AMI)27例的PBMC表面CD44 mRNA进行检测,并以β-actin作为内参进行半定量评价。结果:1UAP和AMI组PBMC表面CD44 mRNA水平均显著高于SAP组和正常对照组(P0.05);2AMI组PBMC表面CD44 mRNA水平显著高于UAP组(P0.05);3SAP组与正常对照组相比,PBMC表面CD44 mRNA水平无统计学差异。结论:PBMC表面CD44表达的水平与冠心病及其病情的严重程度有关联。     

Adenoviral transfer of human interleukin-10 gene in lethal pancreatitis

AIM: To evaluate the therapeutic effect of adenoviral-vector-delivered human interleukin-10 (hIL-10) gene on severe acute pancreatitis (SAP) rats. METHODS: Healthy Sprague-Dawley (SD) rats were intraperitoneally injected with adenoviral IL-10 gene (AdvhIL-10), empty vector (Adv0) or PBS solution. Blood, liver, pancreas and lung were harvested on the second day to examine hIL-10 level by ELISA and serum amylase by enzymatic assay. A SAP model was induced by retrograde injection of sodium taurocholate through pancreatic duct. SAP rats were then administered with AdvhIL-10, Adv0and PBS solution by a single intraperitoneal injection 20 min after SAP induction. In addition to serum amylase assay, levels of hIL-10 and tumor necrosis factor-α (TNF-α) were detected by RT-PCR, ELISA and histological study. The mortality rate was studied and analyzed by Kaplan-Meier and log rank analysis.RESULTS: The levels of hIL-10 in the pancreas, liver and lung of healthy rats increased significantly after AdvhIL-10 injection (1.42 ng/g in liver, 0.91 ng/g in pancreas); while there was no significant change of hIL-10 in the other two control groups. The concentration of hIL-10 was increased significantly in the SAP rats after AdvhIL-10 injection (1.68 ng/g in liver, 1.12 ng/g in pancreas) compared to the other two SAP groups with blank vector or PBS treatment (P&lt;0.05). The serum amylase levels remained normal in the AdvhIL-10 transfected healthy rats. However, the serum amylase level was significantly elevated in the other two control SAP rats. In contrast, serum amylase was down-regulated in the AdvhIL-10 treated SAP groups.The TNF-α expression in the AdvhIL-10 treated SAP rats was significantly lower compared to the other two control SAP groups. The pathohistological changes in the AdvhIL-10 treated group were better than those in the other two control groups. Furthermore, the mortality of the AdvhIL-10 treated group was significantly reduced compared to the other two control groups (P&lt;0.05). CONCLUSION: Adenoviral hIL-10 gene can significantly attenuate the severity of SAP rats, and can be used in the treatment of acute inflammation process.     

An experimental study of emodin assisted early enteral nutrition for the treatment of severe acute pancreatitis

BACKGROUND/AIMS: Both emodin and early enteral nutrition (EEN) have been affirmed as effective means to restore the intestinal mucosal function and abate the severity of severe acute pancreatitis (SAP). However, whether a combined strategy applying both is more effective than either one alone is still undetermined. In this study, we investigated the feasibility and efficacy of emodin assisted early enteral nutrition (EAEEN) for the treatment of SAP. METHODOLOGY: Sixty male Wistar rats were randomly divided into four groups (n=15). SAP was induced in all the rats by a retrograde infusion of 5.0% sodium taurocholate into the pancreatic main duct. Rats in group A received no further intervention, group B with emodin alone, group C with early enteral nutrition (EEN) alone, and group D with emodin assisted early enteral nutrition (EAEEN), respectively, all through an enteral nutrition tube incubated after the induction of SAP. 72 hours after SAP induction, all surviving animals were sacrificed to collect blood and tissue samples for the following measurements: serum amylase, tumor necrosis factor-alpha (TNF-alpha), angiotensin II (AngII) and maleic dialdehyde (MDA), intestinal mucosal secretory IgA (SIgA), pancreatic myeloper oxidase (MPO) activity, and the wet-dry weight ratio of pancreatic tissue (pww/dw). The severity of pancreatic destruction was analyzed by pathological grading and scoring. The severity of intestinal mucosal damage was assessed by the wet-dry weight ratio of ileum (iww/dw), plasma D-lactate and plasma endotoxin. RESULTS: The results of every index in group B, C and D were significantly better than those in group A (P<0.05). Compared with group B and C, group D had significantly reduced levels of serum amylase, TNF-alpha, Ang-II and MDA (P<0.05). Group D also had significantly lowered plasma D-lactate and endotoxin, and decreased pancreatic MPO activity (P<0.05). The pww/dw and iww/dw ratios were decreased, while the SIgA level increased in this group, both with statistical significance (P<0.05). Furthermore, group D had significantly better pancreatic pathologic scores over group B and group C (P<0.05). CONCLUSIONS: Our results suggested that EAEEN could obviously abate the severity of experimental SAP in rats. This combined strategy was rational, safe and more effective than either EEN or emodin used alone, and has a broad potential for future clinical application.