Effects of Propofol on The mRNA Expression of Toll-like Receptor 4 in BV-2 Cells During Mimic Ischemia-Reperfusion Injury In Vitro

This study investigated the effects of propofol on the mRNA expression of Toll-like receptor-4 (TLR4) in BV-2 cells during mimic ischemia-reperfusion (I/R) injury in vitro. BV-2 cells, a mouse microglia line, were cultured and divided into 4 groups at random: control group (group C), ischemia/reperfusion group (group I/R), low-dose propofol (25 μmol/L) intervention group (group PF25) and high-dose propofol (100 μmol/L) intervention group (group PF100). The mRNA expression of TLR4 and NF-κB was measured by means of RT-PCR. TNF-α levels in the supernatants of BV-2 cells were detected by ELISA. The results showed that the mRNA expression of TLR4 and NF-κB was significantly higher in groups I/R, PF25 and PF100 than in group C (P<0.01). And the TNF-α level in the supernatants was elevated in groups I/R, PF25 and PF100 as compared with that in group C (P<0.01). After pre-treatment with propofol, the mRNA expressions of TLR4 and NF-κB and the TNF-α level were significantly decreased in groups PF25 and PF100 in comparison to those in group I/R (P<0.01). And the decrease in those indicators was more significant in group PF100 than in group PF25 (P<0.01). It was concluded that propofol exerted brain-protecting effects during I/R injury by suppressing the mRNA expressions of TLR4 and NF-κB and deceasing the TNF-α level.     

Effects of propofol on the mRNA expression of toll-like receptor 4 in BV-2 cells during mimic ischemia-reperfusion injury <Emphasis Type="Italic">in vitro</Emphasis>

This study investigated the effects of propofol on the mRNA expression of Toll-like receptor-4 （TLR4） in BV-2 cells during mimic ischemia-reperfusion （I/R） injury in vitro. BV-2 cells, a mouse microglia line, were cultured and divided into 4 groups at random： control group （group C）, ischemia/reperfusion group （group I/R）, low-dose propofol （25 μmol/L） intervention group （group PF25） and high-dose propofol （100 μmol/L） intervention group （group PF100）. The mRNA expression of TLR4 and NF-κB was measured by means of RT-PCR. TNF-α levels in the supernatants of BV-2 cells were detected by ELISA. The results showed that the mRNA expression of TLR4 and NF-κB was significantly higher in groups I/R, PF25 and PF100 than in group C （P〈0.01）. And the TNF-α level in the supernatants was elevated in groups I/R, PF25 and PF100 as compared with that in group C （P〈0.01）. After pre-treatment with propofol, the mRNA expressions of TLR4 and NF-κB and the TNF-α level were significantly decreased in groups PF25 and PF100 in comparison to those in group I/R （P〈0.01）. And the decrease in those indicators was more significant in group PF100 than in group PF25 （P〈0.01）. It was concluded that propofol exerted brain-protecting effects during I/R injury by suppressing the mRNA expressions of TLR4 and NF-κB and deceasing the TNF-α level.     

Effect of Wumeiwan on Cytokines TNF-α, IL-6, IL-8, IL-10 and Expression of NF-κBp65 in Rats with Ulcerative Colitis

The effects of Wumeiwan （WMW） on TNF-α, IL-6, IL-8, IL-10 and NF-κBp65 in rats with ulcerative colitis （UC） were investigated, the curative effectiveness of WMW vs salicylazosulfapyridine （SASP） was compared, and the action mechanism was analyzed. Fifty-Six Sprague-Dawley （SD） rats were randomly divided into four groups （n=14 in each group, with equal ratio of male and female）： normal control group, model group, SASP group, and WMW group. Except normal control group, the rat UC models in the remaining three groups were established using the method of 2.4-dinitrochlorobenzene （DNCB） immunization and acetic acid local enema. The rats in model group, SASP group, and WMW group were treated with distilled water, SASP, and WMW respectively. The changes in the symptoms and signs were observed, and levels of IL-6, IL-8, TNF-α, IL-10 and the expression of NF-κBp65 in the colonic tissues were statistically analyzed. The results showed that the levels of IL-6, IL-8, and TNF-α were significantly increased （P〈0.01）, while those of IL-10 significantly reduced （P〈0.01） after establishment of rat UC models as compared with normal control group. The levels of IL-6, IL-8, and TNF-α were obviously lower, but the level of IL-10 was obviously higher in WMW and SASP groups than those in model group （P〈0.05）. The levels of IL-6, IL-8, and TNF-α were lower, while the level oflL-10 was higher in WMW group than in SASP group. NF-κBp65 was expressed negatively or weakly in normal colonic tissues. The positive expression rate of NF-κBp65 in WMW group and SASP group was obviously lower than in model group （P〈0.01）, and there was significant difference between WMW group and SASP group （P〈0.05）. It was concluded that rat UC model was established successfully. WMW could up-regulate the expression of IL-10, down-regulate the expression of TNF-α, IL-6, IL-8, and inhibit the NF-κBp65 activity to adjust immune function, indicating WMW had better curative effects on UC in rats.     

The effect of spinal cord injury on the expression of TGF-β and TNF-α in rat articular cartilage

Objective: To observe the expression of TGF-β and TNF-α in the spinal cord injured rat model and discuss the significance of the articular cartilage metabolism. Methods: 36 SD female rats were randomly divided into 2 groups: Rats models of spinal cord injury were implemented by Allen method. T10 laminectomy was performed in the control group. Both groups of rats were killed respectively in 1w, 3w and 6w. Hematoxylin-eosin stain was given to each slice in the model group and control group. Immunohistochemical stain was applied by using ABC method in the expression of TGF-β and TNF-α. Those expressed level were performed in image analysis and statistics process. Results: TGF-β and TNF-α were mainly distributed on the surface layer of the articular cartilage, with a weak expression in control group. The expression of TNF-α in the model group was more significant than that in the control group in the 1w, and still remained an evident difference with that in control group until the 6w(P ＜ 0.05). TGF-β expression of the model group had no remarkable difference with the control group in the lw (P ＞ 0.05) and prominently became stronger at 6w(P ＜ 0.05). Conclusion: The expression of TNF-o occurred early in the development of spinal cord injury, and the expression of TGF-β became stronger with the revival of spinal neural function. Both expressions were strengthened in articular cartilage in the 3rd week.     

Effect of Combination Regimen of Low-dose Gossypol Acetic Acid with Steroid Hormones on Expression of Protein Kinase C alpha （PKC-tx） and Cyclin D1 in Rat Testes

Objective To investigate the contraceptive mechanism of combination regimen of low-dose gossypol acetic acid (GA) with steroid hormones [desogestre/ethinylestradiol/testosterone undecanote(DSG/EE/TU)]. Methods Adult male rats were randomly divided into four groups. Group GH: rats were fed orally with gossypol acetic acid (GA, 12.5mg/kg) and desogestrel (DSG, 0.125mg/kg)/ ethinylestradiol (EE, 0.025mg/kg)/testosterone undecanoate (TU, 100mg/kg per day, qd×4 weeks or 10 weeks); group G: a single dose of GA (12.5mg/kg per day, qd×4 weeks or 10 weeks); group H: the same dosage of DSG/EE/TU as in group GH; group C: rats were treated with vehicle (1% methyl cellulose) as the control. Expression of protein kinase C alpha (PKC-α) and cyclin D1 in rat testes were tested mainly by immunohistochemistry (IHC) and Western blotting. Results IHC results showed that protein PKC-α was expressed mainly in interstitial tissue of testis among seminiferous tubule. The expression of PKC-α in groups H and GH at week 10 was decreased greatly compared with that in group C. The protein cyclin D1 was expressed mainly in residual body of seminiferous tubule cavosurface and interstitial tissue among seminiferous tubule of testis. Western blotting results showed that the expression of PKC-α in groups H and GH at week 10 was decreased significantly compared with that in group C (P<0.05). The expression of cyclin D1 in groups G, H or GH at week 10 rose significantly compared with that in group C (P<0.05).Conclusion The administration of low-dose gossypol acetic acid with steroid hormones for 10 weeks can decrease the expression of PKC-α greatly.     

Expression of β-1,4-galactosyltransferase I in a surgically-induced rat model of knee osteoarthritic synovitis

Background There are few reports of a biological role for glycosyltransferases in the infiltration of osteoarthritic synovitis. The aim of this research was to investigate the expression and cellular location of β-1,4-galactosyltransferase Ⅰ （β-1,4-GaIT-Ⅰ） in a surgically-induced rat model of knee osteoarthritis （OA）, and explore the role of β-1,4-GalT-Ⅰ in the pathogenesis of OA.Methods Male Sprague-Dawley rats were randomly divided into three groups： OA group, sham group and normal group. The model of OA was established in the right knees of rats by anterior cruciate ligament transaction （ACLT） with partial medial meniscectomy. Fibroblast-like synoviocytes （FLSs） obtained from normal rat synovial tissue were cultured.The expression of β-1,4-GalT-Ⅰ mRNA in the synovial tissue, articular cartilage and FLSs treated with tumor necrosis factor-α （TNF-α） were assayed by real-time PCR. Western-blotting and immunohistochemisty were used to observe the expression of β-1,4-GalT-Ⅰ at the protein level. Double immunofluorescent staining was used to define the location of the β-1,4-GalT-Ⅰ with macrophage-like synoviocytes, FLSs, neutrophils, and TNF-α in the OA synovium. The alteration of TNF-α in FLSs which were treated with lipopolysaccharide （LPS） and β-1,4-GalT-Ⅰ-Ab were detected by enzyme-linked immunosorbent assay （ELISA）.Results The mRNA and protein expression of β-1,4-GalT-Ⅰ increased in synovial tissue of the OA group compared with the normal and sham groups at two and four weeks after the surgery, however, no significant difference appeared in the articular cartilage. Immunohistochemistry also indicated that the β-1,4-GalT-Ⅰ expression in OA synovium at four weeks after surgery increased sharply compared with the control group. β-1,4-GalT-Ⅰ co-localized with macrophage-like synoviocytes, FLSa, neutrophils and TNF-α in rat OA synovitis. Moreover, in vitro β-1,4-GalT-Ⅰ mRNA in FLSs was affected in a dose- and time-dependent manner in response to TNF-α stimulation. ELISA revealed that the expression of TNF-α was attenuated in FLSs in vitro when treated with anti β-1,4-GalT-Ⅰ antibody.Conclusion β-1,4-GalT-Ⅰ may play an important role in the inflammation process of rat OA synovial tissue which would provide the foundation for further researching into the concrete mechanism of β-1,4-GalT-Ⅰ in OA synovitis.     

Expression of Toll-like receptor 4 in neonatal cord blood mononuclear cells in patients with preeclampsia

The expression of Toll-like receptor 4 (TLR4) in neonatal cord blood mononuclear cells (MNCs) and serum TNF-α were investigated in order to explore the roles of TLR4 in the pathogenesis of preeclampsia.The study enrolled 27 patients suffering from preeclampsia (experimental group) and 21 normal pregnancy patients (control group).After MNCs were separated, the expression of TLR4 mRNA and protein was detected by using real-time quantitative PCR and Western blotting respectively, and the expression of TNF-α by using ELISA.The results showed the TLR4 mRNA level in cord blood MNCs (2-CT:0.07±0.17), TLR4 protein expression level (absorbance ratio:0.81%±0.15%) and TNF-α level (9.5±1.73 pg/mL) were all increased in experimental group as compared with control group with the differences being statistically significant (P<0.05).There was a positive correlation between the expression of TLR4 mRNA and TNF-α in both experimental group and control group (r=0.54 and 0.53, respectively, P<0.05).It was concluded that TLR4 expression in the experimental group of cord blood MNCs was increased and there was a positive correlation between the expression of TLR4 mRNA and TNF-α in both groups.TLR4-mediated release of inflammatory cytokines may be one of the important reasons leading to preeclampsia.     

Role of liver X receptors alpha agonist on expressions of LPS-induced inflammatory response associated factor IRAK-4 and NF-kappaB in Kupffer cells

Objective： To explore the role of activated liver X receptor α （LXRα） on the expressions of interleukin-1 receptor associated kinase-4 （IRAK-4） and NF-kappaB （NF-κB） in the inflammatory response which induced by LPS in the Kupffer cells and to investigate the possible mechanisms of LXRα negative regulation of inflammatory response. Methods： The Kupffer cells were isolated from male Kunming mice by collagen perfusion in situ. And these cells were divided into 4 groups： normal control group, LPS treatment group, LXRct agonist T0901317 treatment group, LPS and T0901317 combined treatment group. The LPS treatment group were treated with a final concentration of 1 μg/ml LPS in RPMI 1640 and cultured for 6 h, the T0901317 treatment group were treated with a final concentration of 5 μg/ml in RPMI 1640 and cultured for 24 h, and the combined treatment group received pre-culture for 24 h with a final concentration of 1μg/ml T0901317 in RPMI 1640 and then cultured for 6 h with a final concentration of 5 μg/ml LPS in RPMI 1640. All groups were cultured for 30 h. The expression of LXRα, IRAK-4 and NF-κB at mRNA and protein levels were detected by real-time PCR and Western blotting, and the TNF-α and IL-1β levels were detected by ELISA. Results： The levels of LXRα mRNA and protein were highest in T0901317 group, and lowest in LPS group （P〈0.05）. The level of IRAK4 and NF-κB mRNAs and proteins were evidently lower in the Combined-treated group than in LPS group （P〈0.05）. And the level of TNF-α and IL-1 were observed highest in LPS group （P〈0.05）, but no difference among the Control group, T0901317 group and Combined-treated group （P〉0.05）. Conclusion： These date suggest that the LXR agonists can effectively up-regulate the expressions of LXRα mRNA and protein and inhibit the inflammatory response. This may be via down-regulating the expressions of IRAK4 and NF-κB at mRNA and protein levels.     

Effect of Berberine on Expression of Hepatocyte Nuclear Factor-4α in Rats with Fructose-induced Insulin Resistance

The effects of berberine on the expression of hepatocyte nuclear factor-4α(HNF-4α) in liver of rats with fructose-induced insulin resistance and the molecular mechanism of berberine preventing insulin resistance were investigated.The experimental animals were divided into two groups of 16 animals each.The control group received a control routine diet containing 60% carbohydrate,and the study group a high-fructose diet containing 60% fructose as the sole source of carbohydrate.At the end of 6 weeks these were each subdivided into two groups.One was administered with berberine [187.5 mg/(kg·d) in 5 g/L carboxymethyl cellulose] by intragastric intubation and the other group was treated with a vehicle(5 g/L carboxymethyl cellulose).The rats were fed on the same dietary regimen for the next 4 weeks.After the experimental period of 10 weeks,plasma glucose,insulin and triglyceride levels were measured.HOMA insulin resistance index(HOMA-IR) was assayed.Immunohistochemistry,semiquantitative RT-PCR and western blot were used to detect the expression of HNF-4α in liver.Compared with control diet,fructose feeding induced hyperinsulinemia,HOMA-IR and increased triglyceride(all P<0.01).Berberine prevented the rise in plasma insulin(P<0.01),HOMA-IR(P<0.01) and triglyceride(P<0.05) in the fructose-fed rats.No change in plasma glucose was seen among these groups.The mRNA and protein expression of HNF-4α was decreased in the fructose-fed rats,but berberine could promote its expression.It was concluded that berberine could prevent fructose-induced insulin resistance in rats possibly by promoting the expression HNF-4α in liver.     

The effect of spinal cord injury on the expression of TGF-β and TNF-α in rat articular cartilage

Objective： To observe the expression of TGF-β and TNF-α in the spinal cord injured rat model and discuss the significance of the articular cartilage metabolism. Methods： 36 SD female rats were randomly divided into 2 groups： Rats models of spinal cord injury were implemented by Allen method. T10 laminectomy was performed in the control group. Both groups of rats were killed respectively in 1w, 3w and 6w. Hematoxylin-eosin stain was given to each slice in the model group and control group. Immunohistochemical stain was applied by using ABC method in the expression of TGF-β and TNF-α. Those expressed level were performed in image analysis and statistics process. Results： TGF-β and TNF-α were mainly distributed on the surface layer of the articular cartilage, with a weak expression in control group. The expression of TNF-α in the model group was more significant than that in the control group in the lw, and still remained an evident difference with that in control group until the 6w（P 〈 0.05）. TGF-β expression of the model group had no remarkable difference with the control group in the lw （P 〉 0.05） and prominently became stronger at 6w（P 〈 0.05）. Conclusion： The expression of TNF-α occurred early in the development of spinal cord injury, and the expression of TGF-β became stronger with the revival of spinal neural function. Both expressions were strengthened in articular cartilage in the 3rd week.     

Electroacupuncture Delays Cartilage Degeneration by Modulating Nuclear Factor-κB Signaling Pathway

Objective: To illustrate the molecular mechanisms underlying the therapeutic effects of electroacupuncture(EA) on knee osteoarthritis(OA). Methods: Twenty-seven six-month-old New Zealand white rabbits were allocated into three groups in accordance with a random number table: normal group(no surgery-induced OA; without treatment), model group(surgery-induced OA; without treatment) and EA group [surgery-induced OA; received treatment with EA at acupoints Dubi(ST 35) and Neixiyan(EX-LE 5), 30 min twice a day]. After eight consecutive weeks of treatment, the histopathological alterations in cartilage were observed using optical microscopy and transmission electron microscopy, cartilage degeneration was evaluated by modified Mankin's score principles, the synovial fluid concentration of interleukin-1β(IL-1β), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α) and matrix metalloproteinase-3(MMP-3) were evaluated by enzyme-linked immunosorbent assay, and the protein expression levels of IL-1β, IL-6, TNF-α, MMP-3, IκB kinase-β(IKK-β), nuclear factor of α light polypeptide gene enhancer in B-cells inhibitor α(IκB-α) and nuclear factor-κB(NF-κB) p65 were quantified by Western blot analysis. Results: EA treatment significantly improved cartilage structure arrangement and reduced cellular degeneration. The IL-1β, IL-6, TNF-α and MMP-3 of synovial fluid in the EA-treated group were significantly decreased compared with the model group(all P0.01). Compared with the model group, the IL-1β, IL-6, TNF-α, MMP-3, IKK-β and NF-κB p65 protein expressions in cartilage of EA-treated group were significantly decreased(all P0.01), whereas IκB-α expression was significantly up-regulated(P0.01). Conclusion: EA treatment may delay cartilage degeneration by down-regulating inflammatory factors through NF-κB signaling pathway, which may, in part, explain its clinical efficacy in the treatment of knee OA.     

Huaiqihuang Granules (槐杞黄颗粒) Reduce Proteinuria by Enhancing Nephrin Expression and Regulating Necrosis Factor κB Signaling Pathway in Adriamycin-Induced Nephropathy

Objective: To investigate the effects of Huaiqihuang Granules(槐杞黄颗粒, HQH), a mixture of Chinese herbs including Trametes robiniophila Murr, Fructus Lycii and Polygonatum sibiricum, on adriamycininduced nephropathy(ADRN) in rats and its underlying mechanisms. Methods: Rats with ADRN were divided into four groups: the sham group, the model group(distilled water), the low-dose HQH-treated(2 g/kg) group, and the high-dose HQH-treated(4 g/kg) group. Body weight and 24-h urinary protein(Upro) were checked every week. After 5-week intervention, at the end of the study, the rats were sacrificed and blood samples were collected for examination of biochemical parameters, including glomerular morphological makers, podocyte shape, cellular apoptosis, expressions of nephrin, inflammatory and apoptosis markers. Results: HQH ameliorated the rat's general status, proteinuria, renal morphological appearance and glomerulosclerosis. The decreased expression of nephrin in ADRN rats was increased by HQH, as well as the impaired podocyte foot process fusion. Cytosolic levels of p65 and inhibitor of nuclear factor κBα(IκBα) were decreased in ADRN rats, and recovered by the treatment of HQH. Consistently, the induced expression of tumor necrosis factor α(TNF-α), phosphorylated nuclear factor κB p65(p-NFκB p65) and IκBα in ADRN were markedly suppressed by HQH. In addition, induction of Bax, cleaved caspase-3 and cytochrome C in ADRN rats were suppressed by HQH, indicating the amelioration of apoptosis. Conclusion: HQH could ameliorate renal impairments in ADRN rats by increasing nephrin expression, inhibiting NF-κB signaling pathway via the down-regulation of p-NF-κB p65 and p-IκBα, and suppression of glomerular and tubular apoptosis.     

高温复合脂多糖应激大鼠肾组织TNF-α表达变化

目的 探讨高温与脂多糖(LPS)复合应激大鼠肾组织TNF-α的表达特点.方法 雄性SPF级Wistar大鼠随机分为常温+生理盐水组(C组)、高温+生理盐水组(H组)、常温+LPS组(L组)、高温+LPS组(HL组).置动物于模拟气候舱,HL组、H组暴露环境干球温度(dry bulb temperature,Tdb)为(35.0±0.5)℃,L组和C组Tdb为(26+0.5)℃;HL组和L组动物经尾静脉iv LPS 10 mg/kg,H组和C组动物经尾静脉iv 9g/LNaCl 10 ml/kg.应激120 min时,免疫组织化学SABC染色法检测动物肾组织TNF-α的表达特点,并做常规病理学检查.结果 高温与LPS复合应激组肾组织TNF-α的表达显著增强,肾损伤程度加重.结论 高温与LPS复合应激造成的肾毒性与肾TNF-α表达密切相关.

Abstract：

Objective To investigate the effects of co-exposure of LPS and heat on TNF-α expression in rat kidneys. Methods Male pathogen-free Wistar rats were randomly assigned in saline-injected normothermic control (C group), saline-injected heat exposure (H group), LPS-injected normothermic control (L group), and LPS-injected heat exposure (HL group). The rats in H and HL groups were exposed in a chamber at an ambient dry bulb temperature (Tdb) of 35.0±0.5 degrees, and those in C and L groups were exposed to a Tdb of 26±0.5 degrees. The rats in L and HL groups were given an intravenous injection of LPS (10 mg/kg) via the tail vein to induce endotoxemia, and equivalent normal saline was injected in C and H groups. TNF-α expression in the kidney was detected by immunohistochemical SABC method, and the renal damage was evaluated histologically at 120 min after the treatment. Results Co-exposure of the rats with LPS and heat caused significantly enhanced TNF-α expression and histopathological damage in the kidneys. Conclusion LPS combined with heat exposure causes renal toxicity, while is closely associated with the expression of TNF-α in the kidneys.     

Carboxymethylpachymaran enhances immunologic function of dendritic cells cultured in two kinds of hepatoma carcinoma cell line’s supernatant via nuclear factor κB/Rel pathway

Objective:To study the immunologic function of dendritic cells(DCs) cultured in two kinds of hepatoma cell line’s supernatant and the enhancing effects of carboxymethylpachymaran(CMP) on DCs.Methods:DCs were harvested after stimulation by granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin(IL)-4 from umbilical cord blood using density-gradient centrifugation method. Cultured supernatant of two hepatoma cell lines(HepG2 and HepG2.2.15) were collected for condition medium(CM) according to a volume ratio of supernatant to incomplete RPMI-1640 medium,which was 3:1.CMP was dissolved in incomplete RPMI-1640 medium.Experimental groups were divided according to the culture medium,either CM or with CMP in it.DCs subsets CD83,CD86,CD1a,and d-related human leukocyte antigens(HLA-DR) were analyzed by flow cytometry.The proliferation ability of allogeneic T cells in mixed lymphocyte reaction(MLR) stimulated by DCs was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide(MTT) analysis.IL-12p70,interferon-γ(IFN-γ),and nuclear factor k B(NF- k B) were detected by enzyme-linked immunosorbent assay analysis.Results:The proliferation of lymphocytes and secreting level of IL-12 and expression of phenotype of DCs cultured in two kinds of CM were lower than those of normal group(P<0.01).Compared with the normal group,groups treated with CMP showed a higher expression level of DCs subsets,lymphocyte reproductive activity,as well as IL-12 and IFN-γsecretion levels.Groups treated with CMP also demonstrated higher levels of DCs phenotype expression and IL-12 and IFN-γsecretion in supernatant of MLR and higher lymphocyte reproductive activity compared with CM group(P<0.05).Compared with the normal group,the expression level of NF-k B in DCs nuclear was higher in CMP groups but lower in two CM groups(P<0.05).After CMP was added,the NF-κB expression levels of two CM groups were increased compared with levels before CMP was added(P<0.05).However,there was no significant difference between the two CM groups(P>0.05).Conclusions:Two kinds of hepatoma cell line’s supernatant can inhibit the immunologic function of DCs.This suppressive effect may be related to the inhibition of NF-κB/Rel pathway.CMP may up-regulate the DCs function by activating the NF-κB/Rel pathway.     

Dynamic Observation of Liver Fibrosis in Mice Infected with Schistosoma japonica

Summary： The expression of TNF-α in the liver at different periods post Schistosoma japonica infection and the effect on liver fibrosis after supplementary injection of these cytokines were investigated. The mice infected with schistosome cercariae were divided into 3 groups., normal control group, TNF-α-untreated infection group and TNF-α-treated infection group. ABC immunohistochemistry and pathologic image multimedia quantification system were applied to dynamically detect the activity of TNF-α. The results showed that the levels of TNF-α in the liver in TNF-α-untreated infection group were slowly decreased with prolongation of infection time （from 8th, 11th, 14th to 18th week）, while in the TNF-α-treated infection group, those were increased significantly after intraperitoneal injection of TNF-α at 6th week after infection. At first to 8th week after the final injection of TNF-α, the intrahepatic TNF-α levels in the TNF-α-treated infection group were significantly higher than in the other two groups （P〈0.01）, and the granulomatous inflammation and fibrosis in the liver were also milder in the normal control group. It was concluded that at the early stage of Schistosoma japonica infection mouse liver mainly released Thl cytokine and TNF-α from Thl activated macrophages. Six weeks after infection （post egg deposition）, exogenous supplement with intraperitoneal injection of TNF-α could induce the enhanced expression of Thl cytokines and alleviate the liver granulomatous inflammation and fibrosis.     

Effect of surfactant protein A on lipopolysaccharide-induced tumor necrosis factor-α expression in human proximal tubular epithelial cells

Background Surfactant protein A （SP-A） contributes to the regulation of sepsis-induced acute lung injury.In a previous study,we demonstrated the expression and localization of SP-A in the kidneys.The present study evaluated the effect of SP-A on lipopolysaccharide （LPS）-induced tumor necrosis factor-α （TNF-α） expression and its underlying mechanisms in the human renal tubular epithelial （HK-2） cells.Methods Indirect immunofiuorescence assay was used to detect SP-A distribution and expression in HK-2 cells.HK-2 cells were treated with various concentrations of LPS （0,0.1,1,2,5,and 10 mg/L） for 8 hours and with 5 mg/L LPS for different times （0,2,4,8,16,and 24 hours） to determine the effects of LPS on SP-A and TNF-α expression.Then,HK-2 cells were transfected with SP-A siRNA to analyze nuclear factor κB （NF-κB） P65 and TNF-α expression of HK-2 cells after LPS-treatment.Results Indirect immunofluorescence assay revealed that SP-A is localized to the membrane and cytoplasm of HK-2 cells.Interestingly,SP-A1/SP-A2 and TNF-α expression were found to be significantly increased in HK-2 cells upon LPS treatment.Transfection of LPS-treated HK-2 cells with SP-A siRNA resulted in significant increases in the levels of NF-κB P65 protein and TNF-α mRNA and protein compared to those in non-transfected LPS-treated HK-2 cells.Conclusion SP-A plays an important role in protecting cells against sepsis-induced acute kidney injury by inhibiting NF-κB activity to modulate LPS-induced increase in TNF-α expression.     

Effect of expression of TGF-beta and TNF-alpha with Qidan granule treatment in rats by bleomycin-induced pulmonary fibrosis

Objective: To evaluate the therapeutic effects and mechanisms of Qidan granule in blemycinA5-induced pulmonary interstitial fibrosis (PIF)in rats. Methods: PIF models were established by blemycinA5-induced in rats. They were treated by Qidan granule and Hydrocortisone respectively. The pathological changes and collagen protein disposition were observed, and the expression of TGF-β, TNF-α proteins were measured by immunohistochemical technique . Results: The pulmonary alveolitis and fibrosis were alleviated remarkably in Qidan granule group compared with those in the model control group and hydrocortisone group (P <0. 01). The expression of TGF-β and TNF-α protein were higher in Qidan granule group than those in normal group , and were significantly less than those in the model control group and in hydrocortisone group (P < 0. 01). Conclusion: Qidan granule would ameliorate the pulmonary alveolitis and fibrosis. TGF-β and TNF-α might play an important role in the development of alveolitis and fibro