Protective effect of diallyl trisulfide on liver in rats with sepsis and the mechanism

The protective effects of diallyl trisulfide on liver were examined in rats with sepsis. Sepsis was reproduced in rats by cecum ligation and puncture (CLP). Fifty-six male Wistar rats were randomly divided into sham-operated group (group S, n=8), sepsis model group (group C, n=24), diallyl trisulfide (DATS)-treated group (group D, n=24). Animals in groups C and D were further divided into three subgroups according to different observation time points, with 8 rats in each sub-group.Rats in group D and C were intravenously injected with normal saline or DATS respectively at a dose of 20 mg/kg after the establishment of sepsis model. Eight rats in groups C and D were sacrificed at 3, 6 and 24 h post-CLP and their livers were harvested for detection of interleukin (IL)-1 receptor associated kinase-4 (IRAK-4), nuclear factor-κB (NF-κB), c-fos, c-jun, malondialdehydethhe (MDA) and superoxide dismutase (SOD), tumor necrosis factor alpha (TNF-α) and for pathological examination. The results showed that the levels of serum IRAK-4, NF-κB and TNF-α in hepatic tissues were higher in group C than group S (control group) (P<0.05). After DATS treatment, the levels of IRAK-4 and NF-κB in the hepatic tissues and serum TNF-α in group D were lower than those in group C (P<0.05). The levels of c-fos and c-jun and MDA in the hepatic tissues were higher in group C than in group S (P<0.05). After DATS treatment, the levels of c-fos and c-jun and MDA in the hepatic tissues were significantly lower in group D than in group C (P<0.05). When compared with group S group, concentration of SOD in the hepatic tissues in group C was significantly lower (P<0.05). After DATS treatment, the concentration of SOD in the hepatic tissues was higher in group D than in group C (P<0.05). These findings suggested that treatment with DATS could ameliorate sepsis-induced liver injury in rats. The protective effect might be related to its ability to inhibit the signal pathway of IRAK-4 and NF-κB, thereby decreasing the production o     

Change and significance of nuclear factor-κB in adriamycin induced cardiomyopathy in rats

Background This study aimed at investigating the change and significance of nuclear factor-κB (NF-κB) in eardiomyopathy induced by adriamyein (ADR) in rats. Methods Sixty male Wistar rats were randomly divided into three groups: control, ADR and ADR pyrrolidine dithiocarbamate (PDTC) groups. After 30-day experiment, myocardial histopathological observation was performed. Location and distribution of NF-κB p50 was examined by immunohistochemical assay. Expression of NF-κB p50 protein was examined by immunoboh assay. Electrophoretic Mobility Shift Assay examined activity of NF-κB; Myocardium p53 gene expression was examined by RT-PCR analysis. Results The myocardial lesions of rats were less pronounced in ADR PDTC group than in ADR group. Compared with control group, there were many myocardium nucleuses, which expressed NF-κB p50 and distribute under epicardium. Expression of NF-κB p50 protein in nucleus increased significantly in ADR group. The NF-κB binding activity increased significantly in ADR group. Myocardium expressions of p53 mRNA increased in ADR group. Conclusions The NF-κB binding activity increased significantly in cardiomyopathy induced by ADR in rats. Moreover, NF-κB plays an important role in causing degeneration of myocardial tissue and regulating expression of related-apoptosis genes.     

The effect of root of rhododendron on the activation of NF-κB in a chronic glomerulonephritis rat model

Objective:We have explored the role of nuclear factor kappa B(NF-κB) in the pathogenesis of chronic glomerulonephritis,and investigated the effect of rhododendron root on the activation of NF-κB.Methods:Thirty-six Wistar rats were randomly divided into three groups:a control group,a glomerulonephritis model group and a therapy group(glomerulouephritis animals treated with the root of rhododendron).Bovine serum albumin(BSA) nephritis was induced by subcutaneous immunization and daily intraperitoneal administra-tion of BSA.Twenty-four-hour urinary protein and serum creatinine values were measured,and renal pathology was assessed histologi-cally by optical microscopy and electron microscopy.NF-κB activity was determined by an electrophoretic mobility shift assay(EMSA).Results:Compared with the control rats,glomerulonephrids model rats exhibited a significant increase in both 24 h urinary protein and serum creatinine,and had abnormal renal histology.The administration of the root of rhododendron ameliorated these changes.NF-κ B activity in glomerulonephritis model group was greater than that in rhododendron-treated group,and NF-κB activity was greater in both glomerulonephritis groups than in the control group(P<0.01).Conclusion:These observations suggest that NF-κ B plays a role in the pathogenesis of chronic glomerulonephritis,and rhododendron root may attenuate renal damages by downregulating the activation of NF-kB in this model.     

Effect of Dachengqi Decoction on NF-κB p65 Expression in Lung of Rats with Partial Intestinal Obstruction and the Underlying Mechanism

To investigate the effect of Dachengqi decoction on NF-κB p65 expression in lung of rats with partial intestinal obstruction and the underlying mechanism, 30 SD rats were randomly divided into three groups： sham-operation group, model group and Dachengqi decoction treatment group （Dachengqi group）, with 10 animals in each group. The models were made by partially ligating their large intestines outside the body. The pathological changes were analyzed by HE staining. The expression of NF-κB p65 in rats lung were measured by using real-time polymerase chain reaction and immunohistochemistry respectively. Moreover, the expression of caveolin-1 in rats lung was also measured to. Increased edema, interstitial thickening, hemorrhage, and infiltration of inflammatory cells were found in the model group. In contrast, this change was significantly reduced in Dachengqi group as compared with model group. In addition, the up-regulated caveolin-1 and NF-κB p65 were also suppressed by Dachengqi decoction in lung of rats with partial intestinal obstruction. We are led to concluded that the caveolin-l-NF-κB pathway plays an important role in the development of lung injury of rats with partial intestinal obstruction and Dachengqi decoction could down-regulate the expression of caveolin-1 and NF-κB p65 in lung of rats with partial intestinal obstruction.     

Effect of the regimen of "Gaoshan Hongjingtian" on mechanism of Poly (ADP-ribose) polymerase regulation of nuclear factor kappa B in the experimental diabetic retinopathy

Background Poly (ADP-ribose) polymerase (PARP) plays an important role in the death of retinal capillary cells in diabetic retinopathy (DR), partly via its regulation of nuclear factor kappa B (NF-κB). The current study investigated the effect of the regimen of "Gaoshan Hongjingtian" on the mechanism of PARP regulation of NF-κB, and demonstrated the possible impact of the regimen of "Gaoshan Hongjingtian" and rhodiola sachalinensis on diabetic retinopathy. Methods Wistar rats were made diabetic with streptozotocin, then, were assigned to three groups. After 2 months, these diabetic rats treated with rhodiola sachalinensis or the regimen of “Gaoshan Hongjingtian”, or untreated. Analyses of expression levels of PARP, NF-κB and intercellular adhesion molecule-1 (ICAM-1) on the retinas of rats in different groups were performed by Western blotting and immunohistochemical assays, and mRNA levels of NF-κB and ICAM-1were determined by real-time PCR. In addition, the basement membranes of capillaries in the rats’ retinas were observed by using electron microscopy, and diabetes-induced capillary degeneration (ghost pericytes and acellular capillaries) were quantitated. Results From the third month after the injection, the diabetic rats were given daily with the regimen of “Gaoshan Hongjingtian”, rhodiola sachalinensis or tap water separately. The diabetic rats failed to gain weight when compared with normal age-matched ones, whereas their glycated hemoglobin levels were significantly increased. After 5 months, the mRNA levels of NF-κB and ICAM-1, the protein expression of PARP, NF-κB, and ICAM-1 were significantly increased in the retinas of diabetic rats in untreated group, compared to the nondiabetic controls. After 8 months, the number of degenerated retinal capillaries (ghost pericytes and acellular capillaries) was significantly increased in the diabetic rats in untreated group, compared to normal age-matched rats. The regimen of “Gaoshan Hongjingtian” and rhodiola sachalinensis inhibited diabetes-induced up-expression of PARP, NF-κB, and ICAM-1 in the retinas of diabetic rats with diabetic duration of 5 months. Treatment using the regimen of “Gaoshan Hongjingtian” and rhodiola sachalinensis significangtly inhibited increases in the number of acellular capillaries and pericyte ghosts and suppressed basement membrane thickening in the retinas of rats with diabetes for 8 months, as compared to control diabetic rats. Conclusions These results indicate that PARP plays a role in the pathogenesis of diabetic retinopathy. Rhodiola sachalinensis and the regimen of “Gaoshan Hongjingtian” may have acted on the mechanism of PARP regulation of NF-κB, which suppressed the expression of NF-κB and ICAM-1, and led to inhibition of retinal capillary degeneration.     

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 interleukin-18 on asthmatic airway inflammation and nuclear fa ctor kappa-B in murine models

Objective To investigate the effects of Interleukin-18 (IL-18) on asthmatic airway infla mmation and nuclear factor kappa-B (NF-κB) in a murine asthmatic model. Methods BALB/C mice were randomly divided into three groups: group A(control group,n=10) ; group B (asthmatic model group, n=10); group C (IL-18 injection group, n=10) . The asthmatic model was established in group B and C by respiratory syncytial virus (RSV) killed by ultraviolet light. Saline solution (0.1 ml) and IL-18 (0.1 ml, 1 μg) was injected in groups B and C at seven time points (1, 2, 7, 8 , 9, 21, 22 d). The symptoms and the numbers of eosinophils and plasmacytes in the airways were observed and the expression of NF-κB activation in the lung w as analyzed by Immohistochemistry (IHC) and Western blot. Results The symtoms of group C were more severe than in groups A and B. Group A did no t have EOS and plasmacytes in the airway submucosal while the numbers of eosinop hils ［15±3 (average cell counts per microscopic visual field, the same below) ］ and plasmacytes (10±2) in group B were found to have increased significantly . But the numbers of eosinophils and plasmacytes in group C were decreased sig nificantly when compared with group B (6±2 and 2±1, respectively, both P&lt;0 .05). ISH showed that the expression of NF-κB activation in group B was stro nger than that in groups A and C. The amount of NF-κB inhibitor (IκB) in gro up A and group C were 3.5 times and 2.5 times more than that of group B respec tively via Western blot. Conclusion IL-18 can inhibit asthmatic airway inflammation in mice and its mechamism may b e due to the fact that IL-18 can inhibit the activation of NF-κB in the murin e asthmatic model.     

Pioglitazone ameliorates experimental nonalcoholic steatohepatitis by regulating hepatic nuclear factor-kappa B and cyclooxygenases-2 expression in rats

Background Pioglitazone is effective in nonalcoholic steatohepatitis (NASH), but the mechanism of action is not completely understood. This study was designed to investigate the impact of pioglitazone on hepatic nuclear factor-kappa B (NF-κB) and cyclooxygenases-2 (COX-2) expressions in NASH rats. Methods Thirty Sprague-Dawley male rats were randomly assigned to a control group (n=10), NASH group (n=10) and pioglitazone treatment group (n=10). Liver tissues were processed for histology by HE and Masson stained. Biochemical parameters of antioxidant enzyme activities, tumor necrosis factor alpha (TNF-α), prostaglandin E2 (PGE2) in the serum and hepatic samples were measured. The mRNA and protein expressions of peroxisome proliferator-activated receptor gamma (PPARγ), NF-κB and COX-2 were determined by real-time polymerase chain reaction, Western blotting and immunohistochemistry, respectively. Results There were severe steatosis, moderate inflammatory cellular infiltration and fibrosis in the livers of the NASH models. After treatment, steatosis, inflammation and fibrosis were significantly improved compared with the NASH group (χ2＝20.40, P＜0.001; χ2＝20.17, P＜0.001; χ2＝13.98, P=0.002). The serum and hepatic levels of total anti-oxidation competence (T-AOC), superoxide dismutase (SOD), catalase(CAT), glutathione peroxidase (GSH-PX) and malondialdehyde (MDA) in the NASH group were conspicuous disordered than those indicators in the control group. In the NASH group, the levels of serum TNF-α and PGE2 were significantly increased compared with the control group. Immunohistochemistry showed expressions of NF-κB and COX-2 in livers were significantly elevated, but PPARγ was decreased in the NASH group. Real-time PCR and Western blotting revealed mRNA and protein expressions of COX-2 were increased in the NASH group compared with the control group (0.57±0.08 vs 2.83±0.24; 0.38±0.03 vs 1.00±0.03, P＜0.001 and P=0.004, respectively). After pioglitazone intervention, all of those indicators markedly improved(P＜0.05 or P＜0.01 ). Conclusion Suppressing hepatic NF-κB and COX-2 expression, at least in part, is one of the possible therapeutic mechanisms of pioglitazone in NASH rats.     

Role of β2-adrenoceptor-β-arrestin2-nuclear factor-κB signal transduction pathway and intervention effects of oxymatrine in ulcerative colitis

Objective:To investigate the β2-adrenoceptor(β2AR)-β-arrestin2-nuclear factor-κB(NF-κB) signal transduction pathway and the intervention effects of oxymatrine in a rat model of ulcerative colitis.Methods: Forty SD rats were randomly divided into four groups,which included the normal control group,the model group, the mesalazine group and the oxymatrine treatment group,with 10 rats per group.Experimental colitis induced with trinitrobenzene sulfonic acid(TNBS) was established in each group except the normal control group.The rats in the oxymatrine treatment group were treated with intramuscular injection of oxymatrine 63 mg/(kg·d) for 15 days and the rats in the mesalazine group were treated with mesalazine solution 0.5 g/(kg·d) by gastric lavage for 15 days. The rats in the normal control group and model group were treated with 3 mL water by gastric lavage for 15 days. Diarrhea and bloody stool were carefully observed.Histological changes in colonic tissue were examined on day 7 in 2 rats per group that were randomly selected.The expression of β2AR,β-arrestin2 and NF-κB p65 in colon tissue and spleen lymphocytes were detected with immunohistochemistry and Western immunoblotting techniques on day 16 after fasting for 24 h.Six rats died of lavage with 2 each in the normal control,the model group and the mesalazine group;and were not included in the analysis.Results:The rats in the model group suffered from looser stool and bloody purulent stool after modeling.But in the oxymatrine and mesalazine groups,looser stool and bloody purulent stool reduced after treatment.And the colonic wall in the model group was thickened and the colon length shortened.The colon mucosa was congested in multiple areas with edema,erosion,superficial or linear ulcer and scar formation,while the intestinal mucosa injury reduced in the mesalazine and oxymatrine groups(P<0.01).In colonic mucosa and in spleen lymphocytes,compared with the normal control group,the expression of NF-κBp65 were significantly increased(P<0.01) in the model group while the expressions ofβ2AR andβ-arrestin2 were significantly decreased(P<0.01).Compared with the model group,the expression of NF-κBp65 was significantly decreased in the mesalazine group(P<0.01) and oxymatrine treatment group(P<0.01) while the expressions of β2AR and β-arrestin2 were significantly increased(P<0.01).There were no statistically significant differences in the expression of β2AR,β-arrestin2 and NF-κBp65 between the mesalazine group and oxymatrine group(P>0.05).Conclusions:The β2AR-β-arrestin2-NF-κB signal transduction pathway participated in the pathologic course of ulcerative colitis.Oxymatrine attenuated ulcerative colitis through regulating the β2AR-β-arrestin2-NF-κB signal transduction pathway.     

Effects of Hirudin on High Glucose-Induced Oxidative Stress and Inflammatory Pathway in Rat Dorsal Root Ganglion Neurons

Objective: To investigate protective effects of hirudin on oxidative stress and apoptosis of spinal dorsal root ganglion cells in high-glucose rats at the cellular and molecular level. Methods: Dorsal root ganglion neurons(DRGn) were harvested from embryonic day in 15 SD rats, purified and identificated after primary culture. They were divided into the normal control group, high-glucose(HG) group, positive control(alpha-lipoic acid, ALA) group, low-dose hirudin group(H1), medium-dose hirudin group(H2) and high-dose hirudin group(H3). The control group was cultured by neuron specific culture medium, while the HG group was cultured by neuron specific culture medium and 20 mmol/L glucose(HG medium). The hirudin groups were cultured by HG medium+0.25 IU/mL hirudin(H1), HG medium+0.5 IU/mL hirudin(H2) and HG medium+1 IU/mL hirudin(H3). The ALA group was cultured by HG medium+100 μmol/L ALA. 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenylt etrazolium bromide(MTT) assay was used to explore the optimum concentration and intervention time. Flow cytometry assay was used to detect the level of reactive oxygen series(ROS). Western blot and quantificational realtime polymerase chain reaction(qRT-PCR) were used to detect the expression of protein and mRNA of nuclear factor erythroid 2-related factor 2(Nrf-2), hemeoxygence-1(HO-1), nuclear factor-κB(NF-κB) and Caspase-3. TUNEL assay was used to test the apoptosis rate of different groups. Results: After 24 h of culture, the cell activity of hirudin and ALA groups were higher than that of HG group, and there was a statistical difference between the H1 group and HG group(P0.05). In hirudin groups, the apoptosis rate of cells, the expression of activated Caspase-3 protein and Caspase-3 mRNA were lower than those of HG group(P0.01), higher than those of ALA group(P0.01 or P0.05). The ROS level of hirudin groups was higher than that of ALA group(P0.01), lower than that of HG group(P0.01 or P0.05). The expression of NF-κB(P65) protein in H3 group were lower than those of HG group(P0.05). The expression of Nrf-2 protein in hirudin groups was higher than that of HG group(P0.01), lower than that of ALA group(P0.01 or P0.05). The expression of HO-1 protein in hirudin groups was lower than that of ALA group(P0.01 or P0.05), higher than that of HG group(P0.01 or P0.05). Conclusions: The activity of DRGn cells can be promoted by hirudin under HG conditions. The effects of hirudin on the inhibition of HG on DRGn cells damage mainly include scavenging ROS, up-regulating Nrf-2/HO-1 pathway, inhibiting activation of NF-κB pathway, down-regulating the expression of and Caspase-3 and reducing DRGn cell apoptosis.     

Effect of Chaiqin Chengqi Decoction(柴芩承气汤) on Cholecystokinin Receptor 1-Mediated Signal Transduction of Pancreatic Acinar Cells in Acute Necrotizing Pancreatitis Rats

Objective:To investigate the effect of Chaiqin Chengqi Decoction(柴芩承气汤,CQCQD)on cholecystokinin receptor 1(CCKRI)-mediated signal transduction of pancreatic acinar cell in rats with acute necrotic pancreatitis(ANP).Methods:Twenty-seven Sprague-Dawley rats were randomized into three groups:the control group,the ANP group,and the CQCQD group(9 in each group).ANP rats were induced by two intraperitoneal injections of 8%L-arginine(pH=7.0,4.4 g/kg) over a 2-h period.Rats were treated with 1.5 mL/100 g body weight of CQCQD(CQCQD group) or physiological saline(control and ANP groups) at 2 h interval.And 6 h after induction,pancreatic tissues were collected for histopathological examination.Pancreatic acinar cells were isolated for determination of CCKR1 mRNA and protein expression,phospholipase C(PLC) and inositol-1,4,5-triphosphate(IP3),and determination of fluorescence intensity(Fl)as a measure of intracellular calcium ion concentration[Ca~(2+)]_i.Results:The pancreatic histopathological score(6.2 + 1.1) and the levels of PLC(1,187.2 ±228.2 μg/mL) and IP3(872.2 ±88.4 μg/mL) of acinar cells in the ANP group were higher than those in the control(2.8 ±0.4,682.5 ±121.8 μg/mL,518.4 ±115.8 μg/mL)and the CQCQD(3.8 ±0.8,905.3 ±78.5 μg/mL,611.0 ±42.5 μg/mL) groups(P0.05).[Ca~(2+)]_i Fl for the ANP group(34.8 ±27.0) was higher than that in the control(5.1 ±2.2) and CQCQD(12.6 ±2.5) groups(P0.05).The expression of pancreatic acinar cell CCKR1 mRNA in the ANP group was up-regulated(expression ratio=1.761;P=0.024) compared with the control group.The expression of pancreatic acinar cell CCKR1 mRNA in the CQCQD group was down-regulated(expression ratio=0.311;P=0.035) compared with the ANP group.The ratio of gray values of the CCKR1 and B-actin in the ANP group(1.43 ±0.17) was higher than those in the control(0.70 ±0.15) and CQCQD(0.79 ±0.11) groups(P0.05).Conclusions:Pancreatic acinar cell calcium overload of ANP induced by L-arginine was related to the up-regulated expressions of pancreatic acinar cell CCKR1 mRNA and protein.CQCQD can down-regulate expressions of pancreatic acinar cell CCKR1 mRNA and protein to reduce the PLC and IP3 of pancreatic acinar cells,relieving the calcium overload and reducing the pathological changes in rats with ANP.     

Fuzheng Huayu Formula(扶正化瘀方) Prevents Rat Renal Interstitial Fibrosis Induced by HgCl_2 via Antioxidative Stress and Down-regulation of Nuclear Factor-Kappa B Activity

Objective:To investigate the mechanism of action of Fuzheng Huayu Formula(扶正化瘀方,FZHY)against renal interstitial fibrosis(RIF)relating to oxidative injury and nuclear factor-kappa B(NF-κB)activity.Methods:Thirty-two Sprague-Dawley rats were randomly divided into 3 groups:normal group,model group and FZHY treatment group.The RIF model was induced by oral administration of HgC l2 at a dose of 8 mg/kg body weight once a day for 9 weeks.Meanwhile,rats in FZHY treatment group orally took FZHY at a dose of4.0 g/kg rat weight for 9 weeks.The content of hydroxyproline(Hyp)and collagen deposition in kidney were observed.The activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px),the content of glutathione(GSH)and malondialdehyde(MDA)of kidney were tested.The expressions of inhibitor-κappa B(IκB),phospho-IκB(p-IκB),tumor necrosis factor-α(TNF-α),matrix metalloproteinase-2(MMP-2)andα-smooth muscle actin(α-SMA)were analyzed by Western blot.α-SMA expression was also observed by immunofluorescent staining.MMP-2 activity was measured by gelatin zymography.NF-κB activation was determined by electrophoretic mobility shift assay.Results:Renal interstitial fibrosis was induced by Hg Cl2,demonstrated by remarkably increased Hyp contents and excessive collagen deposition in kidney(P0.01).FZHY significantly inhibited renal interstitial collagen deposition and reduced Hyp content of the Hg Cl2-treated rats(P0.01).GSH content decreased obviously,and MDA content increased significantly in HgC l2-treated rats compared with that of normal rats(P0.01).FZHY significantly increased GSH content and decreased MDA content in the model rats(P0.01).The expressionα-SMA was increased in model rats compared with that of normal rats,FZHY significantly decreased its expression(P0.01).The expressions of p-IκB and TNF-αand MMP-2,MMP-2 activity,and NF-κB activation were increased in model group compared with that in normal group(P0.01),FZHY significantly decreased NF-κB activation,MMP-2 activity and p-IκB and TNF-αexpressions(P0.01).Conclusions:FZHY could protect kidney from oxidative injury intoxicated by Hg Cl2,and antagonized oxidative stress-stimulated NF-κB activity through inhibition of IκB phosphorylation in the interstitial fibrotic kidney,these effects importantly contributed to FZHY action mechanism against renal interstitial fibrosis.     

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.     

Protective Effect of Emodin against Lipopolysaccharidesinduced Corneal Injury in Rats

Objective To investigate the effect of emodin on lipopolysaccharides （LPS）-induced corneal injury in rats. Methods Three parallel incisions on the central surface of corneal epithelium were made and LPS was applied on them to induce corneal injury in Wistar rats. All rats were randomly divided into emodin group （n=40） and keratitis group （n=40）. Rats in the emodin group received subconjunctival injection of emodin and rats in the keratitis group received its vehicle 30 minutes before LPS exposure. At different time points-1 3, 6, 12, and 24 hours after LPS exposure, the symptoms of all rats were observed and the severity of their ocular inflammation was examined with a slit lamp microscope, then 8 rats in each group were killed through cervical dislocation and their eyes were enucleated and prepared to observe pathological changes of corneal tissue under a light microscope. The activation of nuclear factor-loB （NF-κB） under different condi- tions was determined by Western blot. Immunocytochemistry staining with an antibody against intercellular adhesion molecule-1 （ICAM-1） was performed to identify positive cells in corneal tissues. Results The model of acute keratitis was successfully established in Wistar rats. LPS could induce a typical corneal inflammatory response, such as hyperemia, corneal edema and opacity, which were observed in model rats. Compared with keratitis group, both ocular behaviors and damages of the corneal structure were improved in emodin group. Furthermore, the activation of NF-κB and the expression of ICAM-1 induced by LPS were markedly inhibited in emodin group. Conclusion Emodin can inhibit the activation of NF-κB and the expression of ICAM-I induced by LPS in corneas, protect against acute corneal injury, and improve symptoms in rats.