Changes of IFN-γ, IL-6 and TNF-α levels in rats with severe pneumonia

AIM:To study the variety of cytokines in severe bacterial pneumonia of Sprague Dawleg (SD) rats. METHODS:A total of 60 SD rats were randomly divided into three groups: model Ⅰ group (n=24), model Ⅱ group (n=24), and control group (n=12). Rats in the model Ⅰ group and the model Ⅱ group were intratracheally instilled with suspension of klebsiella pneumoniae at different doses. Rats in the control group were intratracheally injected with 1 mL saline. On the 2nd, 4th and 6th day after intratracheal instillation, 1/3 rats in each group were killed to determine the concentration of IFN-γ, IL-6 and TNF-α in blood. RESULTS:The levels of IL-6 and TNF-α in model groups were higher than those in control group, while the level of IFN-γ was lower. The change of cytokines was more significant in the model Ⅱ group (severe pneumonia) than those in the model Ⅰ group. CONCLUSION:The cytokines we studied may play an important role in the pathogenesis of severe pneumonia. The change of cytokines is more significant in severe pneumonia than those in common pneumonia.     

Effects of NF-κB "decoy" oligodeoxynucleotides on TNF-α and IL-6 expression in LPS-induced mouse macrophages

AIM: To study the effect of NF-κB "decoy" oligodeoxynucleotides on TNF-α and IL-6 expression in LPS-induced mouse macrophages. METHODS: Mouse macrophage cell line J774.1 cells were cultured with LPS and liposome-mediated oligodeoxynucleotides, and the levels of TNF-α and IL-6 measured in the different culture supernatant by enzyme linked immunosorbent assay. RNA was extracted from macrophages, and the mRNA expression of TNF-α and IL-6 in macrophages was observed by RT-PCR. RESULTS: NF-κB "decoy" oligodeoxynucleotides decreased the expression of TNF-α and IL-6 in LPS-induced macrophages and inhibited generation of TNF-α and IL-6. The level of TNF-α and IL-6 did not change in control group. CONCLUSIONS: NF-κB "decoy" oligodeoxynucleotides inhibit the expression of TNF-α and IL-6 in LPS-induced macrophages, which is probably due to the specific inhibition of activated NF-κB binding sites .     

Alteration of serum IL-6, IL-8, TNF-α and sICAM-1 in patients with pre-menstruation recurrent aphthous ulceration

AIM:To investigate the serum levels of interleeukin-6(IL-6), interleukin-8(IL-8), tumor necrosis factor-alpha(TNF-α) and soluble intercellular adhesion molecule-1 (sICAM-1)in female patients with pre-menstruation recurrent aphthous ulceration(RAU).METHODS:Serum levels of IL-6, IL-8, TNF-α and sICAM-1 in 21 pre-menstruation RAU patients were examined using ELISA technique, and compared to 10 healthy individuals and 22 the female RAU patients unrelated to menstrual cycle.RESULTS:The serum levels of IL-6, IL-8, TNF-α in patients with pre-menstruation RAU were not only significantly higher than that in the normal control group(P＜0.01), but also higher than that in the RAU patients without pre-menstruous recurrence (P＜0.01).The level of serum TNF-α in the RAU patients without pre-menstruous recurrence was slightly higher than that in the normal control group(P＜0.05).The level of sICAM-1 had not changed.CONCLUSION:The serum levels of IL-6, IL-8 and TNF-α increase in pre-menstruation RAU patients, which might play a role in local lesion of RAU.     

Expression of TNF-α mRNA in hypertrophic myocardium by pressure overload in rats

AIM: To observe the change of TNF-α mRNA in hypertrophic cardiac myocytes induced by pressure overload in rats and the effect of captopril. METHODS: Serum and heart were collected 42 days after the cardiac hypertrophy model made by pressure overload by abdomen aorta-constriction (AC). Hypertrophic parameter and the concentration of TNF-α in serum and left ventricle were determined by ELISA. TNF-α mRNA in cardiac myocytes was determined by in situ hybridization and analyze by ELIA image analysis system. The orientation of TNF-α mRNA in cardiac myocytes was also observed. RESULTS: Left ventricle hypertrophy was observed 42 days after operation. TNF-α mRNA in AC group elevated 98% compared to sham-operated group and descended 64.14% by captopril (P<0.01), but did not descend to the normal level. The expression of TNF-α mRNA showed mostly in myocardial matrix by in situ hybridization. The level of expression was very low in sham-operation group and markedly enhanced after aorta-constriction, but it was decreased when treated by captopril. CONCLUSION: Endogenous TNF-α acts as an important adjustive factor in the pressure overload-induced cardiac hypertrophy and TNF-α mRNA increased in myocardial matrix may be activated by renin-angiotension system.     

Effects of lipopolysaccharide,IL-6 and TNFα on the tissue factor expression of astrocytes

AIM:To study the effects of lipopolysaccharide(LPS), interleukin-6(IL-6)and tumor necrosis factor α (TNFα) on tissue factor(TF) expression of astrocytes. METHODS:Astrocytes were identified with anti-glial fibrillary acidic protein antibody. The TF activity of cell lysate was measured with one stage clotting assay. RESULTS:TF activity of astrocytes of LPS,IL-6,TNFα groups were obviously higher than that of the control group(P <0.05); While LPS,IL-6 and TNFα were combined with trifluoperazine or H₇, their inductive effects were inhibited. CONCLUSION:LPS,IL-6 and TNFα promoted the TF expression of astrocytes and its mechanisms may connected with Calcium/Camodulin and protein kinase C pathway.     

Effects of Radix Angelicae Sinensis on changes of IL-6, TNF-α and bFGF in renal ischemia/reperfusion injury in rabbits

AIM: To determine the effect of Radix Angelicae Sinensis(RAS) on renal ischemia/reperfusion injury in rabbits and to explore its mechanism. METHODS: Twenty-five rabbits were divided randomly into the sham operated group(Control group), renal ischemia/reperfusion injury group(IR group) and RAS+IR group. At the time point of reperfusion 48 h after renal ischemia 1 h, the renal tissue were observed by electron-microscope and the contents of creatinine(Cr) in serum, tumor necrosis factor-α(TNF-α), interleukin-6(IL-6)and basic fibroblast growth factor(bFGF) in the renal tissue were measured. RESULTS: A remarkably degenerative changes in renal tissue were showed under electronmicroscope in IR group, but the changes in RAS+IR group were slight. The contents of Cr, TNF-α and IL-6 in IR group were higher than those in Control group, these parameters in RAS+IR group were lower than those in IR group, the difference between these groups was significant(P< 0.05 or P< 0.01). At the same time, the content of bFGF in IR group was lower than that in Control group(P< 0.01), while the content of bFGF in RAS+IR group was higher than that in IR group(P< 0.01) and Control group(P< 0.05). CONCLUSION: RAS has an effect of alleviating the renal ischemia/reperfusion injury by modulating the production or release of TNF-α, IL-6 and bFGF.     

Pretreatment with external trigeminal nerve electrostimulation inhibits seizures and decreases expression of IL-1β and TNF-α in hippocampus with status epilepticus induced by pentylenetetrazol in rats

AIM:To observe the effect of pretreatment with external trigeminal nerve electrostimulation (eTNS) on behavioral changes and the expression of interleukin-1β (IL-1β) and 　tumor necrosis factor α (TNF-α) in hippocampus of pentylenetetrazol (PTZ)-treated rats. METHODS:The rats were randomly divided into control group, PTZ group and eTNS group, and kindled by PTZ after administered 7 d, 14 d and 28 d of consecutive fake electrostimulation or eTNS. Subsequently, the severity and duration of seizure were quantitatively evaluated. The concentrations of IL-1β and TNF-α in hippocampus were detected by the methods of ELISA and immunohistochemisty. RESULTS:Compared with PTZ group, treatment with eTNS significantly inhibited the severity and duration of seizure (P<0.05), and significantly reduced the content of IL-1β and TNF-α in the hippocampus after status epilepticus (P<0.05 or P<0.01). CONCLUSION:Pretreatment with eTNS may provide a new approach for prevention and treatment of epileptogenesis.     

Effect of TNF-α on osteoporosis pain

AIM: To study the effect of TNF-α on osteoporosis pain. METHODS: Forty female Sparague-Dawley rats, 5 months old, were either sham-operated (sham) or ovariectomized (Ovx), and assigned to sham group (n=16) and Ovx group (n=24). Pain behavioral tests with up-down method and tail withdrawal test were performed once a week beginning at 5th week after the operation. At 8th week after the operation, 8 rats in each group were sacrificed and the L5 dorsal root ganglions (DRG) were collected. The expression of TNF-α in L5 DRG was measured by immunofluorescence. The remaining rats in Ovx group were assigned to 2 subgroups: Ovx-1 and Ovx-2. The rats in Ovx-2 group were administered with thalidomide (an inhibitor of TNF-α synthesis, 50 mg/kg) once a day, while the animals in Ovx-1 group were given same volume of saline. Pain behavioral tests continued to be performed once a week until 10th week after operation. RESULTS: (1)The results of pain behavioral tests showed that the mechanical paw withdrawal threshold and heat tail withdrawal latency in Ovx group at 5th, 6th, 7th and 8th week after surgery were significantly lower than those in sham group (P<0.01). After administration with thalidomide, the mechanical paw withdrawal threshold and heat tail withdrawal latency in Ovx-2 group were restored to the level of sham group, and significantly higher than those in Ovx-1 group (P<0.05). (2)The expression of TNF-α in L5 DRG was significantly higher in Ovx group than that in sham group (P<0.01). CONCLUSION: TNF-α may play an important role in osteoporosis pain by inducing hyperalgesia through taking action on DRG. Inhibition of TNF-α synthesis may relieve menopause osteoporosis pain.     

Characteristics in the secretion of liver TNF-α induced by infusion of LPS into veins in goats

AIM: To study the role of liver in immune regulation in experimental endotoxemia. METHODS: 17 castrated male goats were subjected to simultaneously installing catheters in jugular, hepatic and portal veins by surgery. Four days later, lipopolysaccharide (LPS) was infused in term of three groups as followings: In group ①, LPS of 20 EU (endotoxin unit, EU)·kg^-1 was infused into portal vein; In group ②, LPS of 20 EU·kg^-1 was infused into jugular vein and LPS of 1 500 EU·kg^-1 infused into jugular vein in group ③. Before and after infusion, blood samples were collected from the three veins through the catheters for 8 h.The plasma levels of TNF-α were measured by RIA. RESULTS: In group ①, the plasma TNF-α levels of hepatic and portal vein rose to peak value at 5 h, but that of the jugular vein did not changed. In group ②, the plasma TNF-α levels in hepatic vein rose to peak value at 3 h. The TNF-α levels of jugular vein rose to peak value at 1 h and the one in portal vein enhanced continuously between 0-8 h. In group ③, the plasma TNF-α levels in jugular, hepatic and portal vein rose to significant peaks at 1 h simultaneously. CONCLUSION: During experimental endotoxemia,liver showed different dynamic characteristics in TNF-α secretion according to the pathway and doses of LPS delivery.     

Relationship between levels of IL-2, TNF-α and nitric oxide in aqueous humor after intraocular lens implantation

AIM: To investigate the relationship between the level of interleukin-2 (IL-2), tumour necrosis factor-α (TNF-α) and nitric oxide (NO) in aqueous humor after intraocular lens implantation. METHODS: New Zealand rabbits were divided randomly into three groups: (1) control group; (2) extracapsular cataract extraction group (ECCE); (3) extracapsular cataract extraction and posterior chamber intraocular lens implantation group (ECCE+IOL). The inflammation in all experimental rabbit eyes was observed via zoom-photo slit-lamp microscope on 1, 3, 7, 14 d and 30 d postoperation. Meanwhile, aqueous humor was drawn for white blood cell (WBC) counting and classifying and for determining IL-2, TNF-α and NO₂^-/NO₃^- contents. RESULTS: (1) The level of IL-2 and TNF-α and NO₂^-/ NO₃^- in aqueous humor of ECCE+IOL group were higher than that in ECCE and control at 1 to 14 days postoperation, respectively, it increased to peak value at 3 to 7 days postoperation and decreased gradually two weeks postoperation; (2) The changes in IL-2, TNF-α and NO₂^-/NO₃^- in each group were basically similer; (3) The changes of IL-2 and TNF-α level were closely related with NO content in aqueous humor (r=0.69, P<0.01 and r=0.98, P<0.01). CONCLUSION: IL-2, TNF-α and NO play an important role in intraocular inflammation intraocular lens implantation.     

Relation between TNF-α,TNFR I expression and apoptosis in oral lichen planus

AIM: To examine the expression and distribution of tumor necrosis factor-α (TNF-α), tumor necrosis factor receptor I (TNFR I) and apoptosis in oral lichen planus, and evaluate their roles and relation in the oral lichen. METHODS: Immunohistochemical technique and TUNEL were employed to study the expression of TNF-α, TNFR I and apoptosis in 50 cases of oral lichen planus and 10 normal oral mucosa specimens. RESULTS: Compared with the normal control group, TNF-α expression was upregulated in mononuclear cells in lamina propria and decreased in keratinocytes in oral lichen planus lesion (P<0.05). On the contrary, TNFR I expression was increased in keratinocytes and decreased in lamina propria in oral lichen planus lesion (P<0.05). The increased apoptosis index in keratinocytes and the decreased apoptosis index in lamina propria were found in oral lichen planus (P<0.05). CONCLUSION: The accelerated apoptosis of keratinocytes and the inhibition of lymphocytes apoptosis may contribute to the formation and progression of oral lichen planus.     

Aortic expression of HSP22, TNF-α and eNOS in rats with hyperlipidemia and effects of atorvastatin

AIM:To establish a rat hyperlipidemia model for studying the aortic expression of heat shock protein 22 (HSP22), tumor necrosis factor alpha (TNF-α) and endothelial nitric oxide synthase (eNOS) and the effect of atorvastatin intervention. METHODS:Hyperlipidemia model was established in SD rats. Afterwards, the rats were divided into normal control group, high fat group and high fat+atorvastatin intervention group. The expression of HSP22 and TNF-α in the rat aortas was detected by immunohistochemical assay and the expression of eNOS was assessed by Western blotting. RESULTS:No detectable expression of HSP22 and TNF-α in the normal control group was observed. However, the expression of HSP22 and TNF-α was positive in the high fat group and the atorvastatin intervention group. The mean densities of HSP22 and TNF-α positive particles were significant lower in the atorvastatin intervention group as compared with high fat group (both P＜0.05). The expression of eNOS protein in the high fat group and atorvastatin intervention group was significantly lower than that in normal control group (P<0.01). However, no marked difference of eNOS protein expression between high fat group and atorvastatins intervention group was observed. CONCLUSION: The expression of HSP22 and TNF-α in the rat aortas is increased in the hyperlipidemia rat model. This effect can be restored by atorvastatin treatment. The expression of eNOS in the rat aortas is decreased in the hyperlipidemia rat model, but this tendency could not be attenuated by atorvastatin.     

Effect of oleanolic acid on expression of TNF-α and collagen in silicotic rats in vivo

AIM: To observe the effect of oleanolic acid (OA) on the expression of Tumor necrosis factor-α (TNF-α) and collagen in silicotic rats in vivo and its possible mechanism. METHODS: Male Wistar rats were divided into 4 groups according to the randomized block design: control group, model group, OA group and solvent control group (20 rats in each group). Except control group, the rats in other groups were induced by intratracheal instillation of silicon di-oxide (SiO₂; 250 mg/kg). The rats in OA group were intragastrically administered with OA (60 mg/kg) from the second day of giving SiO₂. The rats in solvent control group were gavaged daily with 0.6% sodium carboxymethyl cellulose solution (10 mL/kg). The rats in control group were given normal saline under the same condition for 56 consecutive days. All rats were killed at the 7th, 14th, 28th and 56th days. The lung coefficient was detected and the morphological changes were observed. The serum contents of TNF-α were detected by ELISA. The content of total collagen in the lung tissue was measured. The protein level of nuclear factor-κB (NF-κB) in the lung tissue was determined by immunohistochemical method. RESULTS: (1) According to the morphological changes, the silicosis model was successfully established. Compared with control group, the lung coefficient and total collagen increased obviously in model group and solvent control group. The lung coefficient and total collagen content in OA group at each time point reduced compared with those in model group and solvent group, and increased compared with those in control group at the corresponding time points. (2) The serum contents of TNF-α in model group and solvent control group significantly increased, peaking at the 14th day, slightly decreasing afterward, and showing statistically significant difference at each time point compared with those in control group. No significant difference between model group and solvent group at different time points was observed. OA had inhibitory effect on the contents of TNF-α compared with model group and solvent group at the corresponding time points. (3) NF-κB in model group and solvent control group significantly increased, peaking at the 28th day, and showing statistically significant difference at each time point compared with those in control group. The NF-κB expression in OA group was similar to model group, but significantly decreased compared with control group at each time point. CONCLUSION: OA inhibits the expression of TNF-α and collagen and attenuates the silicosis fibrosis, which may be related to the NF-κB pathway.     

Effect of aminoguanidine on TNF-α,IL-1β and neuronal apoptosis after focal cerebral ischemic injury in rats

AIM: To evaluate the effect of aminoguanidine (AG) on inflammatory factors and neuronal apoptosis after focal cerebral ischemic injury in rats and the possible mechanism of protective effect of AG against cerebral ischemic injury.METHODS: Thirty male SD rats (weighing 250 g-280 g) were randomly divided into three groups: (1) sham operated group (SH group,n=10),(2) ischemic groups (IS group,n=10),(3) AG group (n=10).In AG group,AG at dose of 100 mg·kg-1 was given intraperitoneally twice a day for 3 consecutive days.In IS group,normal saline was given instead of AG.Focal cerebral ischemia was produced by middle cerebral artery occlusion (MCAO) for 12 h.A nylon thread with rounded tip was inserted into left internal carotid artery cranially until resistance was felt.The distance from bifurcation of common carotid artery to the tip of the thread was about 18-19 mm.Focal cerebral ischemia was confirmed by left Horners syndrome and right side hemiplegia.In SH group,the carotid artery was exposed but no thread was inserted.The expression of tumor necrosis factor-α(TNF-α) was determined by immunochemistry and the content of interleulin-1β(IL-1β) was measured by radioimmunoassay.The expressions of Bcl-2 and Bax protein were detected by flow cytometry.RESULTS: The expression of TNF-α and the content of IL-1β were markedly increased after MCAO.Significantly increased DNA fragmentation,the indication of apoptosis,was detected after MCAO.The expression of TNF-α and the content of IL-1β were significantly lower in AG group than those in IS group.The percentage of apoptosis cells and expression of Bax protein were markedly lower in AG group than those in IS group but still significantly higher than those in SH group.The expression of Bcl-2 protein was markedly higher in AG group than that in IS group.No significant difference in the expression of Bcl-2 protein between IS and SH group was observed.CONCLUSION: AG inhibits the increase in the expression of TNF-α and the content of IL-1β,and protects neurons from apoptosis induced by focal cerebral ischemia through increasing the Bcl-2 protein expression and inhibiting the Bax protein expression.     

Effect of Jiere Xingshen Injection on cAMP,IL-1β content in hypothalamus of ET febrile rabbits

AIM: The effects of Jiere Xingshen(JRXS) Injection on cAMP, IL-1_β content in hypothalamus (HP) of endotoxin(ET)-induced feverish rabbits were studied. METHODS: The ET-induced fever model was established in rabbits and the cAMP content in hypothalamus (HP) and csf, IL-1_β content in HP were determined by radioimmunoassay following intravenous infusion of JRXS. RESULTS: In ET group, the ΔT[(0.40±0.11)℃], TRI₁(1.78±0.79), cAMP content in HP[(2.90±0.40)nmol/g], cAMP content in csf[(0.40±0.11)nmol/L)], IL-1_β content in HP[(6.08±0.79)ng/g] were higher than that of NS and JRXS+ET group (P＜0.01). In JRXS+ET group, the ΔT[(0.10±0.10)℃], TRI₁(0.36±0.64), cAMP content in HP[(1.37±0.27)nmol/g], cAMP content in csf[(14.4±3.69)nmol/L)], IL-1_β content in HP[(2.90±0.37)ng/g] were very close to that of NS group but lower than that of the ET group (P＜0.01);The cAMP content in HP and csf, IL-1_β content in HP paralleled with the fluctuation of temperature. CONCLUTION: JRXS Injection has significant inhibitory effect on ET-induced fever by inhibiting cAMP and IL-1_β production in hypothalamus.     

Alteration of serum interleukin-6 and tumor necrosis factor-α after ischemic stimulation of coronary artery in PTCA

AIM: Inflammatory responses play an important role in the post- percutaneous transluminal coronary angioplasty (PTCA) restenosis and has been demonstrated occuring immediately after PTCA. Interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) are the main inflammatory cytokines. We try to compare the changes in interleukin-6(IL-6) and TNF-α after PTCA in the patients with and without collateral circulation to probe into the pathogenesis of early inflammatory response. METHODS: The extent of myocardial ischemia induced by balloon inflation was quantified by a scoring system referring to the Leaman coronary score. The IL-6、TNF-α levels of coronary heart disease group and control group before and after PTCA are calculated. RESULTS: The concentrations of IL-6 and TNF-α were (9.592±1.847) ng/L and (26.959±1.967) ng/L, respectively, and were significantly increased 4 hours after PTCA. CONCLUSION: IL-6 and TNF-α are sensitive indicators of the early inflammatory response after PTCA. Ischemia scores reflected the extent of ischemia reperfusion injury during PTCA. Collateral circulation decreased the early inflammatory response after PTCA.     

Expression of IL-6 mRNA in endometrium with endometriosis

AIM: The purpose of the present study was to explore the relationship between interleukin-6 mRNA expression and endometriosis. METHODS: Using the rat model, IL-6 mRNA expression in the endometrium was examined by RT-PCR. RESULTS: The expression of IL-6 mRNA in control rats did not change at 2, 4, 6 and 8 weeks after sham operation (P>0.05), but in model rats it gradually increased at 2, 4, 6 and 8 weeks after endometriosis (P<0.01). The expression of IL-6 mRNA in uterine endometrium with endometriosis was lower than in endometriotic tissue, but higher than in endometrium from healthy controls. CONCLUSION: The IL-6 mRNA expression may contribute to the development of endometriosis . The increase in IL-6 mRNA expression may promote the implantation and growth of endometriotic tissue.     

Changes of plasma and myocardial calcitonin gene-related peptide in myocardial stunning rats

AIM: To investigate changes of calcitonin gene-related peptide(CGRP) in myocardial stunning rats. METHODS: Rat in vivo myocardial stunning model was used. CGRP content in plasma and myocardium were determined by radioimmunoassay. RESULTS: Plasma level of CGRP increased significantly (P＜0.01), but in left ventricular myocardium CGRP decreased obviously (P＜0.05) in myocardial stunning group compared with the control group. CONCLUSION: CGRP content in the left ventricular myocardium was negatively correlated with plasma CGRP.