诱导抑郁症患者小胶质细胞炎性反应的机制

小胶质细胞炎性反应是抑郁症患者及相关动物模型中广泛观察到的病理现象,与抑郁症密切相关。抑郁症中诱导小胶质细胞炎性反应的机制包括下丘脑-垂体-肾上腺(HPA)轴激活后糖皮质激素水平变化直接或间接的调控作用、肠道微生物代谢物通过脑肠轴中免疫与神经途径的作用以及损伤相关分子模式(DAMPs)对小胶质细胞的直接激活作用等。

核转录因子-κB反义核酸对肾小球硬化细胞炎性反应的干预

目的检测大鼠肾组织核转录因子-κB(NF-κB),血清IL-6、TNF-α的表达,探讨NF-κB与多柔比星肾病大鼠肾小球细胞炎性反应的关系及反义核酸的干预作用。方法SD雄性大鼠30只分为5组,A组(假手术组),B组(硬化组),C组(正义核酸组),D组(无义核酸组),E组(反义核酸组),每组6只。采用右肾切除加尾静脉注射多柔比星法作肾小球硬化动物模型。第8周各组大鼠予不同药物干预,给药3 d及7 d,分别检测各组大鼠24 h尿蛋白,HE染色观察其肾组织形态学变化,利用图像分析系统计算肾小球硬化指数(GI),应用免疫组织化学、酶联免疫吸附法检测其肾组织NF-κB p65,血清IL-6、TNF-α的表达。采用SPSS 10.0软件进行统计学分析。结果在不同的时间点,E组尿蛋白、GI、NF-κB p65、IL-6、TNF-α的表达与B组比较均显著降低,在7 d时间点,反义核酸的干预效应较3 d时间点强。结论NF-κB与肾小球硬化、细胞炎性反应存在密切关系。NF-κB反义核酸通过肾动脉给药可直接抑制肾组织NF-κB的翻译、表达,抑制肾小球系膜细胞的炎性反应,从而延缓肾小球硬化进程。

巨噬细胞炎性反应蛋白3β复制缺陷型重组腺病毒的构建与鉴定

目的构建小鼠巨噬细胞炎性反应因子-3β(mMIP3β)的复制缺陷型重组腺病毒(Ad-mMIP3β),并在体外表达MIP3β。方法通过在细菌BJ5183内同源重组,利用AdEasyTM系统构建Ad-mMIP3β,经过噬斑筛选、PCR鉴定,然后用CsCl不连续密度梯度离心纯化。利用RT-PCR和Western blot方法研究Ad-mMIP3β的体外表达情况。结果成功构建了MIP3β的复制缺陷型重组腺病毒,并检测到分泌到胞外的MIP3β蛋白。结论MIP3β的复制缺陷型重组腺病毒构建成功,为下一步的肿瘤基因治疗研究奠定了基础。

三种膳食油脂对人血清补体、细胞炎性反应因子和趋化因子及其相关蛋白水平的调节作用及机制初探

目的观察膳食油脂棕榈液油(palm oil,PO)、可可脂(cocoa butter,CO)、大豆油(soybean oil,SO)对人血清补体成分C3、C4、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-10(interleukin-10,IL-10)、正常T细胞表达和分泌的活性调节蛋白(secreted regulatory proteins,RANTES)水平的影响。方法以符合纳入和排除标准的健康青年大学生作为受试者,按体质量指数(body mass index,BMI)采用数字表法随机分为3组,分别是棕榈液油组、可可脂组、大豆油组,每组31名,共93名,分别摄入受试油脂12周,分别在实验开始的第0周和实验结束的第12周检测受试者血清补体成分C3、C4、细胞炎性反应相关因子包括TNF-α、IL-10、趋化因子及RANTES水平。结果各组受试者实验前后比较,受试者血清补体C3、C4水平均降低,差异有统计学意义(P<0.05);棕榈液油组和大豆油组受试者血清TNF-α水平显著性降低(P<0.05);大豆油组受试者血清IL-10水平显著性升高(P<0.05);3组实验前后受试者血清RANTES水平差异无统计学意义(P>0.05)。与实验前比较,实验后,3组油脂血清补体成分、细胞炎性反应因子和活性调节蛋白水平的差异均无统计学意义(P>0.05)。结论棕榈液油、可可脂和大豆油这3种膳食油脂均可能降低人体炎性反应状态,尤其是大豆油的效果更加显著。

盐酸多巴胺注射液联合心脉隆注射液对老年扩张型心肌病中晚期患者心脏功能及心肌细胞炎性反应的影响

目的探究盐酸多巴胺注射液联合心脉隆注射液对老年扩张型心肌病中晚期患者心脏功能及心肌细胞炎性反应的影响。方法选取我科2017年8月至2019年8月129例老年扩张型心肌病中晚期患者作为研究对象,根据随机数字表法分为对照组64例和观察组65例。2组患者均接受常规抗心力衰竭治疗,对照组给予心脉隆注射液治疗,观察组在对照组基础上增加盐酸多巴胺注射液治疗,10 d后,对比2组患者临床疗效、心脏功能:入院24 h及出院时左心室舒张末内径(LVEDd)、左心室射血分数(LVEF)、6 min步行距离(6 MWD)、心脏指数(CI)、心排血量(CO);心肌细胞炎性反应:血清N-末端脑钠肽前体(NT-proBNP)、超敏C反应蛋白(hs-CRP)水平;药物安全性。结果观察组总有效率97%高于对照组73%,LVEF、CI、CO、6MWD高于对照组,NT-proBNP、hs-CRP、LVEDd低于对照组,差异有统计学意义(P<0.05);观察组不良反应发生率为9%,对照组为8%,差异无统计学意义(P>0.05)。结论盐酸多巴胺注射液联合心脉隆注射液治疗老年扩张型心肌病中晚期患者,可提升心脏功能,降低炎性反应,提高临床疗效,药物安全性较高。

肝细胞特异性Sirt1缺失加重小鼠肝脏炎性反应

目的探究沉默信息调节因子1(Sirt1)基因的缺失对小鼠肝脏炎性反应的影响及其作用机制。方法每周记录在高脂喂养下的野生型C57BL/6J雄性小鼠和肝脏特异性Sirt1敲除(Sirt1-LKO)雄性小鼠的体质量,用Western blot和q-PCR检测炎性因子的表达;给小鼠腹腔注射20 mg/kg LPS,用生存曲线来反映小鼠对炎性反应的敏感性。结果与对照组WT小鼠相比,Sirt1-LKO小鼠表现为体质量明显减轻(P<0.05)。另外,生存曲线KaplaneMeier图显示Sirt1-LKO小鼠对LPS刺激呈现低敏感性(P<0.05)。Sirt1-LKO小鼠原代肝细胞中NF-κB蛋白水平增高(P<0.05),及NF-κB下游靶基因炎性因子Tnfα和IL6明显上调(P<0.05)。结论肝细胞特异性Sirt1缺失加重肝脏炎性反应。

Serum soluble interleukin-2 receptor levels in patients with chronic hepatitis B virus infection and its relation with antiHBc

AIM： To investigate the relationship between serum soluble interleukin-2 receptor （sIL-2R） level and anti-HBc in patients with chronic hepatitis B virus （HBV） infection. METHODS： Sera from 100 patients with chronic HBV infection and 30 healthy controls were included in this study. The patients were divided into group A [HBsAg （＋）, HBeAg （＋） and anti-HBc （＋）, n = 50] and group B [HBsAg （＋）, HBeAg （＋） and anti-HBc （-）, n = 50]. sIL-2R levels were determined using ELISA. HBV DNA and alanine aminotransferase （ALT） were also detected. RESULTS： Serum sIL-2R levels were significantly higher in patients with chronic HBV infection than in healthy controls. Moreover, serum sIL-2R levels were significantly higher in patients with HBsAg （＋）, HBeAg （＋） and antiHBc （＋） （976.56±213.51×10^3 U/L） than in patients with HBsAg （＋）, HBeAg （＋） and anti-HBc （-） （393.41±189.54 ×10^3 U/L, P〈 0.01）. A significant relationship was found between serum sIL-2R and ALT levels （P〈 0.01） in patients with chronic HBV infection, but there was no correlation between sIL-2R and HBV DNA levels. The anti-HBc status was significantly related to the age of patients （P〈 0.01）. CONCLUSION： The high sIL-2R level is related to positive anti-HBc in chronic hepatitis B patients. Positive anti-HBc may be related to T-lymphocyte activation and negative anti-HBc may imply immune tolerance in these patients.

Costimulatory molecule programmed death-1 in the cytotoxic response during chronic hepatitis C

Hepatitis C virus（HCV）-specific CD8^＋ T cells play an important role in the resolution of HCV infection. Nevertheless, during chronic hepatitis C these cells lack their effector functions and fail to control the virus. HCV has developed several mechanisms to escape immune control. One of these strategies is the upregulation of negative co-stimulatory molecules such us programmed death-1 （PD-1）. This molecule is upregulated on intrahepatic and peripheral HCV-specific cytotoxic T cells during acute and chronic phases of the disease, whereas PD-1 expression is low in resolved infection. PD-1 expressing HCV-specific CD8^＋ T cells are exhausted with impairment of several effector mechanisms, such as： type-1 cytokine production, expansion ability after antigen encounter and cytotoxic ability. However, PD-1 associated exhaustion can be restored by blocking the interaction between PD-1 and its ligand （PD-L1）. After this blockade, HCV-specific CD8^＋ T cells reacquire their functionality. Nevertheless, functional restoration depends on PD-1 expression level. High PD-l-expressing intrahepatic HCV-specific CD8^＋ T cells do not restore their effector abilities after PD-1/ PD-L1 blockade. The mechanisms by which HCV is able to induce PD-1 up-regulation to escape immune control are unknown. Persistent TCR stimulation by a high level of HCV antigens could favour early PD-1 induction, but the interaction between HCV core protein and gClq receptor could also participate in this process. The PD-1/PD-L1 pathway modulation could be a therapeutic strategy, in conjunction with the regulation of others co-stimulatory pathways, in order to restore immune response aclainst HCV to succeed in clearing the infection.

Leukolysin/MMP25/MT6-MMP: a novel matrix metalloproteinase specifically expressed in the leukocyte lineage

A novel matrix metalloproteinase (MMP) was identified from leukocytes and found to be specifically expressedby peripheral blood leukocytes among 29 different tissuesexamined. Named leukolysin, it encodes for 562 residueswith a conserved MMP structure, i.e., pre-, pro-, catalytic , hinge- and hemopexin-like domains, but also a RXK/RRmotif, known for its role in MMP zymogen activation, anda C-terminal hydrophobic segment. Overall, leukolysindisplays the strongest homology to the newly identifiedMT-MMP subgroup with 45% and 39% identities to MT4and MT1-MMPs vs 30% and 31.5% to MMP1 and 3 respectively. Unlike MT4-MMP whose proteolytic activityremains undefined, a C-terminally truncated leukolysin isexpressed as a strong gelatinolytic species at 28 kDa whichis derived from a cell-associated 34 kDa proenzyme, presumably by furin or proprotein convertase mediated removal of the propeptide (-6 kDa). By green fluorescentprotein (GFP) tagging, the intracellular proenzyme is localized to granules throughout the cell, suggesting thatactivation occur immediately prior to secretion. Taken together, leukolysin may be part of the proteolytic arsenaldeployed by leukocytes during inflammatory responses.

Mesenchymal stem cells： a new strategy for immunosuppression and tissue repair

Mesenchymal stem cells （MSCs） have great potential for treating various diseases, especially those related to tissue damage involving immune reactions. Various studies have demonstrated that MSCs are strongly immunosuppressive in vitro and in vivo. Our recent studies have shown that un-stimulated MSCs are indeed incapable of immunosuppression; they become potently immunosuppressive upon stimulation with the supernatant of activated lymphocytes, or with combinations of IFN-γ, with TNF-α, IL-1α or IL-1β. This observation revealed that under certain circumstances, inflammatory cytokines can actually become immunosuppressive. We showed that there is a species variation in the mechanisms of MSC-mediated immunosuppression： immunosuppression by cytokine-primed mouse MSCs is mediated by nitric oxide （NO）, whereas immunosuppression by cytokine-primed human MSCs is executed through indoleamine 2, 3-dioxygenase （IDO）. Additionally, upon stimulation with the inflammatory cytokines, both mouse and human MSCs secrete several leukocyte chemokines that apparently serve to attract immune cells into the proximity with MSCs, where NO or IDO is predicted to be most active. Therefore, immunosuppression by inflammatory cytokine-stimulated MSCs occurs via the concerted action of chemokines and immune-inhibitory NO or IDO produced by MSCs. Thus, our results provide novel information about the mechanisms of MSC-mediated immunosuppression and for better application of MSCs in treating tissue injuries induced by immune responses.

Lactobacillus crispatus M206119 exacerbates murine DSS-colitis by interfering with inflammatory responses

AIM： TO investigate the role of Lactobacillus crispatus （L. crispatus） strain China Center for Type Culture Col- lection （CCTCC） M206119 in intestinal inflammation.METHODS： Forty 8-wk-old Balb/c mice （20± 2 g） were divided into four groups of 10 mice each. Three groups that had received dextran sulfate sodium （DSS） were administered normal saline, sulfasalazine or CCTCC M206119 strain, and the fourth group received none of these. We assessed the severity of colitis using a disease activity index, measured the colon length and weight, collected stools and mesenteric lymph nodes for bacterial microflora analysis. One centimeter of the proximal colon, middle colon and distal colon were collected and fixed in 10% buffered formalin, dehydrated in ethanol, and embedded in paraffin. Interleukin （IL）- 1β, IL-6 and tumor necrosis factor （TNF）-α expression was detected using reverse transcription polymerase chain reaction. Protective factors zonula occludens （ZO）-1 and β-defensin 2 were detected by immunoblot-ting. The features of CCTCC M206119 strain were identified based on morphology, biochemical profile, and 16S RNA sequencing.RESULTS： DSS-colitis animals treated with CCTCC M206119 had markedly more severe disease, with greater weight loss, diarrhea, fecal bleeding, and shortened colon length. In addition, the CCTCC-M206119- treated group had comparatively higher histologi- cal scores and more neutrophil infiltration than the controls. Expression of protective factors ZO-1 and β-defensin 2 was downregulated due to destruction of the mucosal barrier after CCTCC M206119 strain treatment. An in vitro assay demonstrated that CCTCC M206119 strain increased the nuclear translocation of nuclear factor-κB in epithelial cells. Intestinal proinflam- matory or anti-inflammatory cytokine responses were evaluated. Proinflammatory colonic cytokine （IL-Iβ, IL-6 and TNF-α） levels were clearly increased in CCTCC- M206119-treated animals, whereas anti-inflammatory colonic cytokine （IL-10） level was lowered compared with saline or 5-aminosalicylic-acid-treated DSS-colitis mice. Next, CCTCC M206119 strain was characterized as 1. crispatus by microscopic morphology, biochemical tests and 16S rRNA gene level.CONCLUSION： Not all lactobacilli are beneficial for in- testinal inflammation, and L. crispatus CCTCC M206119 strain is involved in exacerbation of intestinal inflamma- tion in DSS-colitis mice.

Lipopolysaccharide induced hyper- and hypo-responsiveness in macrophage cell lines

Objective: To build a cell model of LPS-induced hyper- and hypo-responsiveness in macrophage cells . Methods: Macrophage cell line RAW264. 7 was pre-cultured with or without 10 ng/ml LPS for 18 h, then challenged with lipopolysaccharide(LPS) , or MDP, Zymosan, PAF, FMLP, PMA for 24 h. The levels of TNF-α , IL-1 , IL-6, IL-10 , NO and O2-, were measured. Results: LI'S pretreatment markedly inhibited TNF-a NO and IL-6 production, but increased IL-1, IL-10 and O2- release to LPS challenge. LPS pretreatment also altered macrophage responsiveness to the other stimuli. Conclusion: LPS can induce hyper- and hypo-responsiveness simultaneously in the macrophage cell lines. Changes in macrophage responsiveness depend on stimuli and effectors which are measured.

The roles of toll-like receptors in carcinogenesis and cancer immunotherapy

Toll-like receptors （TLRs） are probably the most important class of pattern-recognition receptors. Members of the TLR family play key roles in the both innate and adaptive immune responses. Recognition of pathogen-associated molecular patterns （PAMPs） by TLRs, either alone or in heterodimedzation with other TLR or non-TLR receptors, induces the production of signals that are responsible for the activation of genes important for an effective host defense, especially those of proinflammatory cytokines. Thus, TLRs are involved in the development of many pathological conditions including infectious diseases, tissue damage, and cancer especially. In this review, the contribution of TLRs to tumorgenesis is evaluated. We hope to provide new insight into the progression of cancer and more importantly into the potential for TLRs as targets of therapeutics.

TaqMan real-time fluorescent quantitative RT-PCR in detection of macrophage inflammatory protein-2γ mRNA in myocarditis murine

Objective: To study the role of macrophage inflammatory protein (MIP)-2γ in myocarditis pathogenesis in BALB/c mice. Methods: The relationship between the progression of Coxsarckie virus B3(CVB3) viral myocarditis and experimental autoimmune myocarditis and MIP-2γ mRNA expression in mouse was studied by TaqMan real-time fluorescent quantitative RT-PCR. Results: MIP-2γ mRNA expression rose on 3 to 5 d after CVB3 infection, reached peak on 7 d, and returned to normal level until 14 d, which corresponded well with the disease course. The MIP-2γ mRNA expression level rose significantly on the day 18 d after immunization with porcine cardiac myosin, which was consistent with pathological examination. Conclusion: MIP-2γ may be involved in the pathogenesis of myocarditis.

Effects of Resveratrol on the Proliferation and Apoptosis in Synoviocytes of Rheumatoid Arthritis

This study was undertaken to investigate the regulatory effect of Resveratrol (Res) on the proliferation and apop- tosis of synoviocytes of patients with rheumatoid arthritis (RA), as the proliferation of synoviocytes of patients was deter- mined by MTT chromatometry and the apoptosis of these cells was assayed with TUNEL flow cytometry. It was found in this experiment that the degree of cell proliferation of the Res-treated group with dosages of 50-400 μM was significantly reduced in comparison with that of the control group, but percentage of the apoptotic cells demonstrated with TUNEL labeling was el- evated under treatment with Res at the same dosages in a concentration-dependent manner. The difference between the Res- treated group and the control group was quite significant ( P < 0.01). It is concluded that Res shows a potent anti-prolifera- tive effect on synoviocytes of patients with RA with induction of cell apoptosis, and it is likely a valuable candidate for the chemotherapy and management of patients with RA.

Effects of recombinant sCR1 on the immune inflammatory reaction in acute spinal cord injury tissue of rats

Objective: To determine the effects of recombinant soluble complement receptor type I (sCR1) on the immune inflammatory reaction in acute spinal cord injury tissue of rats and its protective effects. Methods: SD rat models of acute spinal cord injury were prepared by modified Allen’s method. The motor function of the rat lower extremities in sCR1 group and normal saline (NS) group was evaluated by the tiltboard experiment at 12 h, 1 d, 3 d, 7 d, and 14 d. The neutrophil infiltration and C3c positive expression were observed. The myeloperoxidase activity was assessed in the injury tissue at 12 h, 1 d, 3 d, 7 d, and 14 d after injury in the two groups. Results: The motor function of rat in sCR1 group at 3 d, 7 d, and 14 d was obviously better than that in NS group (P< 0.01, P< 0.01, P< 0.01). C3c positive expression in sCR1 group at each time point after injury was obviously less than that in NS group (P< 0.01). The myeloperoxidase activity in sCR1 group at each time point after injury was obviously less than that in NS group (P< 0.01). Conclusions: Recombinant soluble complement receptor type I (sCR1) can lessen the immune inflammatory reaction in acute spinal cord injury tissue and relieve secondary spinal cord injury by inhibiting the activation of the complement system.

Effects of phospholipase D on cardiopulmonary bypass-induced neutrophil priming

Objective:To investigate the relationship betwe en phospholipase D (PLD) activation and neutrophil priming induced by cardiopulm onary bypass (CPB), and try to clarify whether CPB-induced systemic inflammator y response can be attenuated by inhibiting neutrophilic PLD activation. Methods:Neutrophils were isolated from arterial blood of 8 pat ients undergoing valve replacement before operation and 30 min after initiation of CPB respectively. Both the preoperative and CPB-stirred neutrophils were sub divided into 5 groups by receiving different experimental interventions: (1) bac terial lipopolysaccharide (LPS, 10 ng·ml -1 ), (2) N-formylmethionylph enylalanine (fMLP, 1 μmol·L -1 ), (3) LPS+fMLP, (4) 1-butanol ( 0.5 %)+ LPS+fMLP, (5) vehicle. Elastase and myeloperoxidase (MPO) release was measured f or the parameters of neutrophil activation, neutrophil PLD activity was determin ed by quantitation of choline produced from the stable product of phosphatidylch oline catalyzed by PLD. Results:(1) Preoperative neutrophils treated with LPS+fMLP pre sented significantly higher PLD activity ( 13.48 ± 2.61 nmol choline·h -1 ·mg -1 ) and released more elastase and MPO than cells treated with v ehicle (PLD activity 3.70 ± 0.49 nmol choline·h -1 ·mg -1 , P< 0.01 ), LPS (P< 0.01 ) and fMLP respectively. In 1-butanol+LPS+fMLP group, PLD activity of preoperative neutrop hils was lower than that in LPS+fMLP group (P< 0.01 ), b esides the release of elastase and MPO decreased sharply below both LPS + fMLP a nd fMLP groups (P< 0.01 ). In LPS group, PLD activity wa s higher (P< 0.01 ), while elastase and MPO release did not differ from control. fMLP group presented PLD activity, elastase and MPO rel ease higher than control (P< 0.01 ); nevertheless, lower than LPS+fMLP group (P< 0.01 ). (2) CPB-stirred neutro phils presented prominent PLD activity increment, and even the control level was 3.59 -fold of the pre-operative control (P< 0.01 ) . PLD activity in LPS+fMLP group was higher than that in other groups. Notably, PLD activity was even nonstatistically lower in 1-butanol+LPS+fMLP group than t hat in LPS or fMLP group. CPB-stirred neutrophils in LPS+fMLP group released mo re elastase and MPO than control, LPS, and 1-butanol+LPS+fMLP groups did ( P< 0.01 ); however, neither of the release was statistically different from that of fMLP group. Conclusions:Cardiopulmonary bypass enables neutrophil priming accompanied with significant increase in PLD activity. Inhibition of neutrophil PLD activation attenuates its priming and may alleviate CPB-induced systemic in flammatory reaction.