血管内皮细胞间连接蛋白VE-cadherin和β-连环蛋白在颅内动脉瘤中的表达

目的 观察血管内皮钙黏蛋白(VE-cadherin)和β-连环蛋白(β-catenin)在SD大鼠和人颅内动脉瘤壁中的表达,探讨血管内皮细胞连接受损与颅内动脉瘤形成的关系.方法 建立肾性高血压诱导的SD大鼠颅内动脉瘤模型,获取8个SD大鼠颅内动脉瘤.另取15例颅内动脉瘤组织标本,应用免疫组织化学方法检测VE-cadherin在大鼠脑动脉瘤壁中的表达,以及VE-cadherin和β-catenin在人脑动脉瘤壁的表达.按照Martina的方法对血管内皮层免疫反应强度进行评分.结果 正常SD大鼠和人的脑动脉血管壁上内皮层完整,VE-cadherin呈棕褐色线样均匀表达.SD大鼠颅内动脉瘤壁上VE-cadherin表达减少,呈间断性非均匀分布.正常SD大鼠脑动脉血管壁和颅内动脉瘤壁免疫反应强度评分分别为2.40 ±0.55和1.38±0.51,两组间差异有统计学意义(t=3.41,P＜0.01).人颅内动脉瘤壁内皮排列紊乱,VE-cadherin和β-catenin表达也明显减少,分布不均匀或表达缺失.正常脑动脉与颅内动脉瘤VE-cadherin和β-catenin免疫反应强度评分分别为2.60±0.54/1.33±0.49和2.80±0.45/1.33±0.49,VE-cadherin和β-catenin在两者间差异均有统计学意义(t分别为4.88和5.91;P＜0.01).结论 颅内动脉瘤的形成过程中存在内皮细胞连接受损.

Abstract：

Objective To verify whether vascular endothelial intercellular gap injury takes part in the formation of intracranial aneurysms by assaying the expression of vascular endothelial intercellular gap proteins VE-cadherin and β-catenin in the rat experimental intracranial aneurysms and human intracranial aneurysms. Methods Twenty Sprague-Dawley rats were used for establishing a model of hypertension-induced experimental intracranial aneurysm. Eight aneurysmal changes were found in the junction of the anterior cerebralartery (ACA) and the olfactort artery (OA) of rats, and immunohistochemistry was performed to detect the expression level of VE-cadherin. The expression of VE-cadherin and β-catenin was also assayed by using immunohistochemistry in fifteen human intracranial aneurysms. Results The VE-cadherin expression was reduced obviously in the intracranial aneurysms of rats as compared with the nomal cerebral artery ( 1.38 ±0. 51 vs 2.40 ±0. 55 ,P ＜0. 01 ). In human intracranial aneurysms, the expression of VE-cadherin was lower than that in the human superficial temporal artery ( 1.33 ± 0. 49 vs 2. 60 ± 0. 54, P ＜0. 01 ). And the expression of β-catenin was also reduced obviously as compared with the controls ( 1.33 ±0. 49 vs 2. 80 ± 0. 45 ,P ＜0. 01 ). Conclusion Vascular endothelial intercellular gap injury may involve in the formation of intracranial aneurysms.     

脾动脉缩窄对门静脉高压大鼠脾脏iNOS表达及Th1/Th2平衡的影响

Objective To investigate the effect and the potential mechanism of splenic artery coarctation on the expression of iNOS and Th1/Th2 cytokines in spleen of cirrhotic rats with portal hypertension (PHT). Methods Cirrhotic rats were randomized into 3 groups (n= 10):sham operation group (SOG), splenic artery coarctation group (SAC) and splenic artery ligation group (SAL). Ten normal rats treated with sham operation were employed to serve as normal control group (NCG). Immunohistochemial staining was used to observe iNOS. RT-PCR was used to detect IFN-γ and IL-4mRNA. The Pearson's correlation analysis was used to investigate the relationship between iNOS and IFN-γ or IL-4. Results The expression of iNOS was increased significantly in spleen of cirrhotic rats as compared with NCG(P＜0. 01). It was decreased after SAC and SAL compared with SOG (P＜0. 01). The expression of IFN-γmRNA and IFN-γ/IL-4 of SOG were decreased but IL-4mRNA increased significantly than that of NCG(P＜0.01). IFN-γmRNA was increased after SAC compared with SOG (P＜0.05). IL-4mRNA was decreased and IFN-γ/IL-4 increased after SAC and SAL compared with SOG (P＜0. 05). The expression of iNOS was negatively correlated with the expression of IFN-γmRNA(r=-0.672, P＜ 0.01 ) and positively correlated with the expression of IL-4 mRNA (r=0.634,P＜0. 01). Conclusion The expression of iNOS is decreased and IFN-γ/IL-4 increased after SAC in spleen of cirrhotic rats with PHT and it may improve Th1/Th2 polarization by reducing the expression of iNOS.     

脾动脉缩窄对门静脉高压大鼠脾脏iNOS表达及Th1/Th2平衡的影响

Objective To investigate the effect and the potential mechanism of splenic artery coarctation on the expression of iNOS and Th1/Th2 cytokines in spleen of cirrhotic rats with portal hypertension (PHT). Methods Cirrhotic rats were randomized into 3 groups (n= 10):sham operation group (SOG), splenic artery coarctation group (SAC) and splenic artery ligation group (SAL). Ten normal rats treated with sham operation were employed to serve as normal control group (NCG). Immunohistochemial staining was used to observe iNOS. RT-PCR was used to detect IFN-γ and IL-4mRNA. The Pearson's correlation analysis was used to investigate the relationship between iNOS and IFN-γ or IL-4. Results The expression of iNOS was increased significantly in spleen of cirrhotic rats as compared with NCG(P＜0. 01). It was decreased after SAC and SAL compared with SOG (P＜0. 01). The expression of IFN-γmRNA and IFN-γ/IL-4 of SOG were decreased but IL-4mRNA increased significantly than that of NCG(P＜0.01). IFN-γmRNA was increased after SAC compared with SOG (P＜0.05). IL-4mRNA was decreased and IFN-γ/IL-4 increased after SAC and SAL compared with SOG (P＜0. 05). The expression of iNOS was negatively correlated with the expression of IFN-γmRNA(r=-0.672, P＜ 0.01 ) and positively correlated with the expression of IL-4 mRNA (r=0.634,P＜0. 01). Conclusion The expression of iNOS is decreased and IFN-γ/IL-4 increased after SAC in spleen of cirrhotic rats with PHT and it may improve Th1/Th2 polarization by reducing the expression of iNOS.     

The dynamic balance of T-bet+VGATA-3+ and ROR-γt+ VFOXP3 + cells in rat liver allograft according to acute rejection severity

Objective To evaluate the dynamic balance of Th1, Th2, Th17 and iTreg cells in the process of acute rejection (AR) , orthotopic liver transplantation (OLT) was performed from LEW rats to BN rats. Methods OLT donated by male inbred LEW rats was performed on male inbred BN rats by Kamada' s two-cuff technique without liver arterial anastomosis, AR severity was graded using the Banff schema ,and the immunohistological method was applied to detect the expression of T-bet+, GATA-3+, RORγt+, and FOXP3+ cells in liver allografts. Results The number of T-bet+ cells (Control group: 19. 3±5. 1;Mild AR group: 63. 7±9. 7;Moderate AR group: 40. 9±13. 6;Severe AR group: 32. 3±3. 3) and RORγt + (Control group: 6. 5±1. 4;Mild AR group: 6.4±3. 1;Moderate AR group: 23. 2±9. 1;Severe AR group: 42. 6±14. 1) was significantly increased in mild, moderate and severe AR groups as compared with control group (all P<0.01). The number of GATA-3+cells (Control group: 7. 1±1. 3;Mild AR group: 13. 5±4. 8;Moderate AR group: 23. 2±9. 1;Severe AR group: 42. 6± 14. 1) showed no significant difference between mild AR group and control group (P>0. 05) , while it was significantly higher in moderate and severe AR group (P<0. 05 and P<0. 01). The number of FOXP3 + cells ( Control group;9.5±2.4;Mild AR group: 13. 5±4.8;Moderate AR group: 24.5 ±4.9;Severe AR group: 39.3±11.7) had no statistically significant difference (P>0. 05), but it was significantly decreased in moderate and severe AR group (all P<0.01). The ratio of T-bet + cells/GATA-3+cells was more significantly associated with AR than that of RORγt+cells/FOXP3+ cells in the early stage. Conclusion There is dynamic change in T-het+,GATA-3+,RORγt+and FOXP3+cells in the liver allografts.The T-bet+,GATA-3,RORγt'and FOXP3+cells were involved in AR,and the ratio of T-bet+cell/sGATA-3+cells was more significantly associated with AR than that of RORγt+cells/FOXP3+cells in the early stage.     

The dynamic balance of T-bet+VGATA-3+ and ROR-γt+ VFOXP3 + cells in rat liver allograft according to acute rejection severity

Objective To evaluate the dynamic balance of Th1, Th2, Th17 and iTreg cells in the process of acute rejection (AR) , orthotopic liver transplantation (OLT) was performed from LEW rats to BN rats. Methods OLT donated by male inbred LEW rats was performed on male inbred BN rats by Kamada' s two-cuff technique without liver arterial anastomosis, AR severity was graded using the Banff schema ,and the immunohistological method was applied to detect the expression of T-bet+, GATA-3+, RORγt+, and FOXP3+ cells in liver allografts. Results The number of T-bet+ cells (Control group: 19. 3±5. 1;Mild AR group: 63. 7±9. 7;Moderate AR group: 40. 9±13. 6;Severe AR group: 32. 3±3. 3) and RORγt + (Control group: 6. 5±1. 4;Mild AR group: 6.4±3. 1;Moderate AR group: 23. 2±9. 1;Severe AR group: 42. 6±14. 1) was significantly increased in mild, moderate and severe AR groups as compared with control group (all P<0.01). The number of GATA-3+cells (Control group: 7. 1±1. 3;Mild AR group: 13. 5±4. 8;Moderate AR group: 23. 2±9. 1;Severe AR group: 42. 6± 14. 1) showed no significant difference between mild AR group and control group (P>0. 05) , while it was significantly higher in moderate and severe AR group (P<0. 05 and P<0. 01). The number of FOXP3 + cells ( Control group;9.5±2.4;Mild AR group: 13. 5±4.8;Moderate AR group: 24.5 ±4.9;Severe AR group: 39.3±11.7) had no statistically significant difference (P>0. 05), but it was significantly decreased in moderate and severe AR group (all P<0.01). The ratio of T-bet + cells/GATA-3+cells was more significantly associated with AR than that of RORγt+cells/FOXP3+ cells in the early stage. Conclusion There is dynamic change in T-het+,GATA-3+,RORγt+and FOXP3+cells in the liver allografts.The T-bet+,GATA-3,RORγt'and FOXP3+cells were involved in AR,and the ratio of T-bet+cell/sGATA-3+cells was more significantly associated with AR than that of RORγt+cells/FOXP3+cells in the early stage.     

颅内外血管搭桥治疗复杂动脉瘤及搭桥血管闭塞的防治

目的 探讨颅内外血管搭桥技术在复杂动脉瘤治疗中的应用和移植血管闭塞的原因.方法 回顾分析2006年11月至2008年11月以颅内外血管搭桥治疗的20例颅内复杂动脉瘤,并总结大隐静脉闭塞的原因.其中男性8例,女性12例,年龄27～65岁,平均54.5岁.动脉瘤位于海绵窦内13例,床突旁动脉瘤4例,床突上动脉瘤2例,基底动脉干动脉瘤1例;动脉瘤＜15 mm者4例,15～25 mm者12例,＞25 mm者4例.结果 1例术中出现血管闭塞,1例术后出现血管闭塞,出院时格拉斯哥预后评分(GOS)4～5分者17例(17/20),3分(重残)2例(2/20),1分(死亡)1例(1/20).19例生存患者术后6个月随访GOS评分4～5分者18例(18/19),3分(重残)1例(1/19).结论 颅内外血管搭桥技术治疗颅内复杂动脉瘤是有效、安全的,机械因素和血流动力学因素是移植血管闭塞最主要的原因,移植血管的长期通畅率还有待进一步观察.

Abstract：

Objectives To study the revascularization techniques in the treatment of intracranial complex aneurysms and occlusive reasons of bypass vessel. Methods The 20 complex intracranial aneurysms who underwent saphenous bypass treatment from November 2006 to November 2008 were retrospectively analyzed and the occlusive reasons were studied. Of the 20 patients, 12 were female and 8 were male, mean age was 54.5 years (ranged 27-65 years). The distribution of the lesions inclued 13 cavenous sinus aneurysms, 4 paraclinoidal aneurysms, 2 supraclinoidal aneurysms, and 1 basilar trunk aneurysm. Four aneurysms were giant (diameter ＞25 mm), 12 aneurysms were large( 15-25 mm), and 4 aneurysms were medium( 10-15 mm) size. Results One saphenous vein was occlused intraoperativly and one saphenous vein was occlused postoperativly. At discharge, 18 out of the 20 patients had Glasgow Outcome Scale (GOS) score of 4 orS, 2 patients had score of 3, and 1 patient had score of 1. At 6 months follow up, 18 of 19 survivors had GOS score of : or 5 and 1 patient had score of 3. Conclusions Extracranial-intracranial revascularization technique is a safe and effective method in the treatment of complex aneurysms. Mechanical and hemodynamic factors are two leading reasons for occlusion of bypass vessels. Long-term bypass vessels patent rate still needs further observation.     

Effect of local testicular heating on spermatogenic cell apoptosis in rats

Aim: To study the effect of testicular local heating on spermatoge-nic cell apoptosis in rats. Methods: Seventy male SD rats were divided into the heat-treated and the control groups. The former was exposed to heat (43 ℃) for 12 hours. Each group was further divided into seven subgroups with respect to the time of observation after heat exposure, i.e., 12 h and 1 days, 3 days, 6 days, 10 days, 50 days and 80 days, respectively. In each subgroup, sper-matogenic cell apoptosis was examined by means of electron microscopy, flow cytometry and terminal deoxynucleotidyl trans-ferase-mediated dUDP-nick end labeling (TUNEL) methods. Results: The percentage of cells with sub-haploid and the percentage of positive TUNEL cells were significantly higher in the heat-treated groups than in the controls (P<0.01). The reaction of cell apoptosis to local heat was highly selective: spermatocytes were the most sensitive, followed by spermatids, spermatozoa and sper-matogonia in a decreasing order. Conclusion: Local testic     

与大鼠胰岛细胞联合移植的骨髓间充质干细胞的免疫调节作用

目的 研究同种异体或自体的大鼠骨髓间充质干细胞(MSC)与胰岛肝内联合移植对胰岛移植物的免疫调节作用及其机制.方法 以链佐星制备Lewis大鼠的糖尿病模型,作为胰岛移植受者,分为3组:单纯移植组大鼠经门静脉单独移植SD大鼠胰岛6000 IEQ/kg;同系MSC组大鼠经门静脉共同移植1×109/L的Lewis大鼠MSC 1 ml与SD大鼠胰岛6000 IEQ/kg;同种MSC组大鼠经门静脉共同移植1×109/L的SD大鼠MSC 1 ml与SD大鼠胰岛6000 IEQ/kg.检测受鼠的血糖变化,术后1、3 d大鼠外周血γ干扰素(IFN-γ)、白细胞介素(IL)-2、IL-4和IL-10的含量.结果 3组大鼠术后第1天血糖均下降到13.9 mmol/L以下.同系MSC组移植物存活时间为(11.38±4.03)d,同种MSC组为(5.50±2.07)d,单纯移植组为(2.88±1.25)d(P＜0.01).术后1、3 d,单纯移植组IFN-γ和IL-2的含量显著高于同系MSC组和同种MSC组(P＜0.01),同种MSC组IFN-γ和IL-2的含量高于同系MSC组(P＜0.05);单纯移植组IL-10的含量低于同系MSC组和同种MSC组(P＜0.01),同系MSC组IL-10的含量与司种MSC组相比较,差异无统计学意义(P＞0.05);各组IL-4含量的差异无统计学意义(P＞0.05).结论 MSC与同种胰岛共移植可以延长胰岛移植物存活时间,应用同系MSC的效果优于同种异体MSC.共移植的MSC主要通过减少TH1类细胞因子(IFN-y和IL-2)的表达使受者TH1/TH2平衡向TH2方向偏移.

Abstract：

Objective To compare the immune regulation of syngenic and allogenic mesenchymal stem cells (MSCs) in the transplantation combined with islets. Methods After induction of diabetes in 30 Lewis rats with streptozotocin (STZ), the recipient Lewis rats received islets from SD rats combined with syngenic (group B) or allogenic (group C) MSCs injection via the portal vein. The group of islets transplanted alone served as control (group A). The survival time of grafts in all groups was assessed by the level of blood glucose. ELISA was used to detect the levels of interferon-γ (IFN-γ), interleukin 2 (IL-2), IL-4 and IL-10 in the peripheral blood on the 1st and 3rd day after transplantation. Results The blood glucose levels in all three groups were decreased in a normal range (13. 9 mmol/L) and the survival time of grafts in group B (11.38 ± 4. 03 days) was significantly longer than in group C (5. 50± 2. 07 days) as well as group A (2. 88 ± 1.25 days). On the 1 st and 3rd day after transplantation, the levels of TH 1 cytokines IFN-γ and IL-2 in group A were significantly higher than in groups C and B (P＜0.05). Meanwhile the levels of TH 2 cytokine IL-10were increased in group B, but there was no significant difference between groups A and C (P＞0.05). The levels of IL-4 had no significant difference among these three groups (P ＞ 0.05).Conclusion Islet transplantation combined with MSCs could prolong the survival time of grafts.Syngenic MSCs, superior to allogenic ones, were more effective in changing the balance of TH1/TH2to TH2. Decreased expression of TH1 cytokine (IFN-γ, IL-2), which was closely related to the induction of immune tolerance, was beneficial to the long-term survival of grafts.     

静脉注射后脂肪来源干细胞在创伤大鼠体内的迁移

Objective To explore the migration of intravenously injected adipose tissue-derived stem cells(ADSCs) in SD rats with soft tissue wound. Methods 1.8cm×1.8cm full thickness skin defect and 0.5cm in depth soft tissue defects were made on the back of 6 SD rats. ADSCs were isolated and cultured in vitro for 3 passages. 2.4×106 ADSCs were labeled with DiI and transplanted into the SD rats through taft vein. Nounal skin and wound tissue Samples were collected for fluorescent distribution observation 24、48 days after injection, respectively. Results Compared to normal skin, more florescent positive cells were detected in the margin and deep layer of the wound 24 days after operation. But it is accumulated within dermis and adenoid 48 days after operation. Conclusion Wound can probably induce the migration and accumulation of intravenously transplanted ADSCs.     

鞘内联合应用白细胞介素-1受体拮抗剂、氯胺酮对慢性神经病理性疼痛的影响

objective To observe the effect of intrathecal injection of IL-1ra and ketamine for chronic constriction injury (CCI)in rats. Methods 40 male SD rats were randomly divided into 5 groups: Sham operation group;CCI group; IL-1ra group;ketamine group; IL-1ra and ketamine group. Thermal withdrawal latency (TWL) and mechanical withdrawal threshold(MWT) of 2 h pre -drug and 2,4,6 h after injected drugs at day 21 in rats were performed by Hargreave's thermal radiation apparatus and von Frey's method of micro filament, respectively. The rats were killed after the behavior test was finished. The expression of p-ERK was measured in the spinal cord by immunohistochemical analysis and western blot method. Results The group of combined injection IL-1rα and ketamine produced significantly more potent analgesia compared with that of CCI group. MWT(7.1±2.2 )and TWL ( 13.5 ±3.6)were increased from 2 hours to 6 hours after intrathecal injection of IL-1rα and ketamine(P<0.01 ). The expressions of p-ERK and the number of positive cells in the combined injection group were significantly lower than that of CCI group (P<0.01). The effect of IL-1ra and ketamine in lower dosages is tendency enhanced than that of higher dosage alone. Conclusion The combined injection of IL-lra and ketamine in lower dosage can produce synergistic abirritation in chronic pain.     

Relationship between T-helper cell 17/regulatory T cell imbalance and atherosclerosis cardiovascular disease in maintenance hemodialysis patients

Objective To investigate whether the T-helper cell 17 (Th17)/regulatory T cell (Treg) of patients on maintenance hemodialysis (MHD) present imbalance in terms of proportion and function, and if so, the relationship between such imbalance and atherosclerosis cardiovascular disease (ASCVD). Methods Fifty-seven MHD patients, included 25 with ASCVD and 32 without ASCVD, and 24 healthy volunteers were enrolled. The common carotid artery intima media thickness (CCA-IMT), carotid plaque and plaque area were determined with high-resolution B-mode ultrasonography. Treg (CD4+CD25+Foxp3+) and Th17 (CD4+IL-17+) were measured by flow cytometry. The Foxp3 and RAR-related orphan receptor γt (RORγt) mRNA expressions were measured by real-time PCR. TGF-β1, IL-10, IL-17 and IL-6 in serum were detected by ELISA. Results There were decreased Treg proportion, Foxp3 mRNA, TGF-β1 and IL-10, and increased Th17 proportion, RORγt mRNA, IL-17, IL-6 and Th17/Treg in the ASCVD group compared with that in the non-ASCVD group and healthy group (all P＜0.01). No correlation was observed between Treg and CCA-IMT, but IL-10 were negatively correlated with CCA-IMT (P＜0.05). Th17, IL-17 and ratio of Th17/Treg were positively correlated with CCA-IMT (all P＜0.05). MHD patients with carotid plaques had lower Treg, TGF-β1 and IL-10, higher Th17, IL-17 and ratio of Th17/Treg than those without carotid plaques (all P＜0.05). Moreover, Treg proportion was negatively correlated with carotid plaque area in MHD patients with carotid plaques (P＜0.01). Conclusions The Th17/Treg numerical and functional imbalance exists in MHD patients, especially in patients with ASCVD. This might act synergistically with micro-inflammation on immune-mediated atherosclerosis and contribute to the high incidence of ASCVD.     

脾动脉缩窄对门静脉高压大鼠脾脏iNOS表达及Th1/Th2平衡的影响

目的 观察脾动脉缩窄对肝硬化门静脉高压大鼠脾脏iNOS、Th1/Th2型细胞因子表达的影响,并探讨机制.方法 肝硬化门静脉高压大鼠随机分3组(n=10):假手术组(SOG)、脾动脉缩窄术组(SAC)和脾动脉结扎术组(SAL);正常大鼠10只行假手术作为对照组(NCG).免疫组化法测脾脏iNOS表达,RT-PCR法测脾脏IFN-γ、IL-4mRNA表达,对iNOS与IFN-γ、IL-4表达量作相关分析.结果 SOG脾脏iNOS明显高于NCG(P＜0.01),SAC和SAL明显低于SOG(P＜0.01).SOG脾脏IFN-γmRNA和IFN-γ/IL-4明显低于NCG(P＜0.01),IL-4mRNA明显高于NCG(P＜0.01);SAC脾脏IFN-γmRNA高于SOG(P＜0.05),SAC和SAL脾脏IL-4mRNA低于SOG(P＜0.05),而IFN-γ/IL-4高于SOG(P＜0.05).iNOS与IFN-γ负相关(r=-0.672,P＜0.01),与IL-4正相关(r=0.634,P＜0.01).结论 脾动脉缩窄术后门静脉高压大鼠脾脏iNOS表达降低,IFN-γ/IL-4升高,脾脏Th1/Th2失衡改善可能与术后iNOS表达降低有关.     

大鼠肝移植物中T-bet+/GATA-3+和ROR-γt+/FOXP3+细胞随排斥程度的动态变化及意义

目的 检测Th1、Th2、Th17和iTreg CD4+T细胞亚系特征性表达的核转录因子以探究它们在大鼠肝移植急性排斥中的动态变化及意义.方法 Kamada"二袖套"法建立大鼠肝移植急性排斥模型,按Banff标准对排斥程度进行分级;免疫组织化学方法检测Th1、Th2、Th17和iTreg特征性表达的核转录因子,对阳性细胞计数.结果 与对照组比较,轻度排斥组、中度排斥组和重度排斥组中T-bet+(对照组:19.3±5.1;轻度排斥组:63.7±9.7;中度排斥组:40.9±13.6;重度排斥组:32.3±3.3)细胞数和RORγt+(对照组:6.5±1.4;轻度排斥组:8.3±2.3;中度排斥组:26.8±3.2;重度排斥组:39.2±12.8)细胞数显著升高(P<0.01);轻度排斥组中GATA-3+(对照组:7.1±1.3;轻度排斥组:6.4±3.1;中度排斥组:23.2±9.1;重度排斥组:42.6±14.1)细胞数无明显升高(P>0.05),中度排斥组中明显升高(P>0.05),重度排斥组显著升高(P<0.01);轻度排斥组FOXP3+(对照组:9.5±2.4;轻度排斥组:13.5±4.8;中度排斥组:24.5±4.9;重度排斥组:39.3±11.7)细胞数无明显升高(P>0.05),中度、重度排斥组显著升高(P<0.01).结论 大鼠肝移植术后急性排斥的发生与T-bet+、GATA-3+、RORγt+和FOXP3+细胞相关;在排斥早期,T-bet+/GATA-3+平衡与排斥的发生相关性比RORγt+/FOXP3+平衡更强.     

芦荟多糖调控Th17/Treg细胞平衡缓解肠炎的实验研究

目的探讨芦荟提取液芦荟多糖是否通过调节辅助性T细胞（Th17）与调节性T细胞（Treg）免疫失衡,缓解2,4,6-三硝基苯磺酸（2,4,6-TNBS）诱导的肠炎。 方法用完全随机数字法将30只SD大鼠分成正常组（PBS液灌肠）、芦荟多糖治疗组（TNBS-乙醇液灌肠+肛门注入芦荟多糖）及模型组（TNBS-乙醇液灌肠）,每组10只。记录大鼠体质量变化、疾病活动指数（DAI）。取结肠组织进行宏、微观损伤评分及病理学检查;检测结肠组织髓过氧化物酶（MPO）活性以及细胞因子转化生长因子（TGF-β）、白细胞介素（IL）-10、IL-17A、IL-6 mRNA的表达,转录因子RORγt、Foxp3⁺及信号转导和转录激活因子（STAT）3、5蛋白表达;检测血浆中细胞因子IL-1β、肿瘤坏死因子α（TNF-α）、肿瘤坏死因子α刺激基因6（TSG-6）的表达。 结果与模型组比较,芦荟多糖治疗组的DAI、MPO、组织病理学评分及血浆细胞因子IL-1β、TNF-α的蛋白表达显著降低,血浆TSG-6浓度增加。芦荟多糖抑制了结肠组织中促炎细胞因子IL-17A、IL-6 mRNA表达,以及转录因子RORγt活化及STAT3磷酸化,促进了抑炎细胞因子IL-10、TGF-βmRNA表达,以及Foxp3⁺表达及STAT5磷酸化。 结论芦荟多糖可以通过纠正Th17/Treg细胞失衡,缓解TNBS诱导的实验性肠炎。     

低剂量雷公藤甲素预处理减轻小鼠肝脏缺血再灌注损伤的作用及其机制

目的 探讨雷公藤甲素(TPT)预处理减轻小鼠肝脏缺血再灌注损伤的作用及其作用机制.方法 将60只雄性C57BL/6小鼠分成4组,每组15只.(1)TPT手术组:手术方法参照Kobayashi法,术中夹闭肝门静脉左支、肝动脉左支及左肝管,90min后开放血管,建立小鼠肝脏缺血再灌注损伤模型;(2)TPT假手术组:手术方式同TPT手术组,但术中不阻断血流,仅用生理盐水(NS)纱布覆盖切口90 min;(3)NS手术组:手术方式同TPT手术组;(4)NS假手术组:手术方式同TPT假手术组.两TPT组小鼠于术前1周开始腹腔注射TPT0.1 mg·kg-1·d-1,术前1 h加用1次,而两NS组同期仅腹腔注射等体积无菌NS.再灌注后24 h,检测各组小鼠肝功能和观察肝组织病理学变化,采用流式细胞术检测各组TH17细胞占单个核细胞的比例,采用实时聚合酶链反应(PCR)检测各组肝组织中IL-17和ROR-γt mRNA的表达,采用酶联免疫吸附试验(ELISA)检测血清中IL-6、IL-17和TGF-β的含量.结果 TPT手术组肝功能的损伤较NS手术组明显减轻(P＜0.05);NS假手术组、TPT假手术组、NS手术组和TPT手术组TH17细胞占单个核细胞的比例分别为(0.72±00.23)%、(0.41±0.18)%、(4.26±0.82)%和(1.77±0.53)%;两TPT组IL-17和ROR-γt mRNA的表达量以及外周血中IL-6、IL-17和TGF-β的含量,均明显低于相应的NS组(P＜0.05).结论 低剂量TPT预处理能够减轻小鼠肝脏缺血再灌注损伤,这可能与TPT预处理抑制小鼠体内Th17细胞分化、发育及其功能有关.     

Th17 alloimmunity prevents neonatal establishment of lymphoid chimerism in IL-4-deprived mice

Immune responses in newborn mice are known to be biased toward the helper type 2 phenotype. This may account for their propensity to develop tolerance. Herein, we evaluated the effects of IL-4 deprivation on CD4(+) T-cell activities elicited by neonatal exposure to allogeneic spleen cells. We showed that chimerism, Th2-type polarization and pathology, as well as skin allograft acceptance were inhibited in BALB/c mice immunized at birth with (A/J x BALB/c) F(1) spleen cells upon in vivo IL-4 neutralization. While IL-4 neutralization inhibited the development of Th2 cells in this model, it led to the accumulation of IL-17A, IL-17F, IL-22, IL-6 and RORγt mRNA in the spleen or graft tissues. Moreover, IL-4 deprivation led to the differentiation of donor-specific Th17 cells with a concomitant Th1 response characterized by IFN-γ production. The Th17-type response emerging in IL-4-deprived mice was found to mediate both intragraft neutrophil infiltration and the abrogation of B-cell chimerism. Neutralization of this Th17 response failed however to restore functional skin graft acceptance. Collectively, our observations indicate that the neonatal Th2 response opposes the development of Th17 cells, and that Th17 cells are responsible for controlling lymphoid chimerism in mice neonatally injected with semiallogeneic cells.     

Dynamic changes in Th1, Th17, and FoxP3+ T cells in patients with acute cellular rejection after cardiac transplantation

Previously, studies suggest that CD4(+) effector T-cell subsets participate in allograft rejection. However, the dynamic changes and relative roles of these CD4(+) effector T-cell subsets, especially Th17 cells, have not been systemically examined in patients with acute rejection after cardiac transplantation. In this study, we have studied and compared these CD4(+) T-cell subsets in peripheral blood and endomyocardial biopsies (EMB) in patients with stable-graft and acute cellular rejection. We observed that the gene expressions including T-bet, IFN-γ, RORγt, IL-17, IL-23, and FoxP3, the functional marker of Th1, Th17, and FoxP3(+) CD4(+) T cells, were elevated in EMB samples from patients with acute graft rejection. Accordingly, the percentages of circulating Th1, Th17, and FoxP3(+) CD4(+) T cells were also significantly increased. The data suggest that Th1, Th17, and FoxP3(+) CD4(+) T cells are associated with acute graft rejection in patients with cardiac transplantation.