Effects of TOLL-like receptor 4 on passively sensitized human airway smooth muscle cells proliferation and synthesis and secretion function of TGF-beta1 / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the activation of Toll like receptor 4 (TLR4) on passively sensitized human airway smooth muscle cells (HASMCs) proliferation and the synthesis and secretion function.</p><p><b>METHODS</b>Through the cultivation of primary HASMCs, we studied TLR4 expression on cell surface, cell proliferation and transformation of parturient factor-beta1 (TGF-beta1) in asthma under the condition of synthesis and secretion level by passively sensitized HASMCs with asthma serum.</p><p><b>RESULTS</b>Compared with the control group, in passive sensitized group and TNF-alpha group TLR4 expression were significantly increased (P < 0.01), significantly enhanced proliferation (P < 0.01), total protein concentration, IgE secretion and TGF-beta1 were significantly higher (P < 0.01); and all the above parameters were increased more significantly in TNF group compared with those in the target effect of passively group; and those parameters were significantly reduced in anti-TLR4 antibody group compared with those in the target effect both of passively sensitized group and TNF-alpha group.</p><p><b>CONCLUSION</b>TLR4 on passively sensitized HASMCs activated can induce the excessive proliferation of HASMCs and a large number of synthesis and secretion of TGF-beta1, resulting in changing airway micro-environment, which involved in airway remodeling in asthma.</p>

The effect of LPS on airway inflammation, airway remodeling and TLR4 expression in asthmatic rat / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effects of lipopolysaccharide (LPS) on airway inflammation, airway remodeling and the expression of Toll-like receptor 4 (TLR4) mRNA in asthmatic rats.</p><p><b>METHODS</b>Twenty-four SPF level SD rats were randomly divided into four groups (n = 6): control group, low dose of LPS group, high dose of LPS group and asthma group. Using ovalbumin (OVA) to sensitize and challenge to establish asthmatic rat model. Observed pathological changes of lung tissue by HE staining, inflammatory cell infiltration was observed by airway wall eosinophils (EOS) counts; airway resistance was determined; image analysis software was used to determine the thickness of airway wall, detected airway smooth muscle TLR4 expression levels by RT-PCR.</p><p><b>RESULTS</b>The rat airway resistance and the EOS number of airway wall and the thickness of airway wall in asthma group, low dose of LPS group and high dose of LPS group were significantly higher than those in control group (P < 0.01). The above-mentioned parameters of high dose of LPS group showed significantly lower than those in asthma group and low dose of LPS group (P < 0.05). The expression of rat airway smooth muscle TLR4 mRNA in low dose of LPS group and high dose of LPS group were significantly higher than those in asthma group (P < 0.01). And the expression of rat airway smooth muscle TLR4 mRNA in high dose of LPS group was significantly higher than that in low dose of LPS group (P < 0.05).</p><p><b>CONCLUSION</b>TLR4 plays an important role in asthmatic airway inflammation and airway remodeling, LPS may play double-sided regulation in asthmatic airway inflammation and airway remodeling by activated TLR4.</p>

Expression of Cysteinyl leukotriene receptor 1 in bronchial epithelial cell regulated by tumor necrosis factor-α / 中华实用儿科临床杂志

Objective To explore whether TNF-α involves in the modulation of Cysteinyl leukotriene receptor 1 (CysLT1) expression in bronchial epithelial cells.Methods The bronchial epithelial cell lines 16HBE cells were stimulated with different concentration (0.00,0.05,0.50,5.00,20.00 μg/L) of TNF-α for 48 hours,and CysLT1 mRNA in 16HBE cells was measured by reverse transcription(RT)-PCR.CysLT1 expression was detected by immunohistochemistry.Results 16HBE cells did not express CysLT1,after the cells were treated with TNF-α,obvious expression of CysLT1 were detected by immunohistochemistry.The weak CysLT1 mRNA expression was observed by RT-PCR in 16HBE cells,and after the cells were treated with TNF-α for 48 hours,CysLT1 mRNA expression were upregulated.When the concentrations of TNF-α were 0.00,0.05,0.50,5.00,and 20.00 μg/L respectively,the relative intensities of CysLT1 mRNA/β-actin were 0.048,0.105,0.177,0.182,0.495,respectively.Conclusions TNF-α can upregulate CysLT1 mRNA expression in 16HBE ceils in a dose-dependent manner.When infected by virus,respiratory tract produces abundant TNF-α.The TNF-α can upregulate the expression of CysLT1 in epithelial cells,enhance inflammation reaction in respiratory tract.This may explain partially the mechanism of exacerbation of asthma induced by respiratory tract infection.

Effect of TLR4 on the migration of asthmatic airway smooth muscle cells induced by airway epithelial cells / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the effect of Toll-like receptor 4 (TLR4) activation on the migration of asthmatic airway smooth muscle cell (ASMCs) induced by airway epithelial cells.</p><p><b>METHODS</b>Primary ASMCs were cultured by the method of cell digestion. Cell culture supernatant of RTE cells were collected by TNF-alpha stimulation of epithelial cells. Detected the IL-8 and RANTES levels in the supernatant. The transmembrane migration of asthmatic ASMCs were detected by Modified Boyden chemotaxis chamber. The effect of TLR4 on the migration of asthmatic ASMCs induced by epithelial cells with TLR4 antibody drugs as a tool.</p><p><b>RESULTS</b>The levels of IL-8 and RANTES in the supernatant of TNF-alpha groups were significantly increased, and that in the 20 ng/ml group was significantly higher than other groups (P < 0.01). The transmembrane migration of asthmatic ASMCs groups was increased than that of control group. The transmembrane migration of asthmatic ASMCs from asthma group and TNF-alpha + TLR4 antibody group was significantly decreased compared with that in TNF-alpha group (P < 0.01). The migration of asthma ASMCs from TNF-alpha + TLR4 antibody group was increased than that of asthma group (P < 0.05).</p><p><b>CONCLUSION</b>TLR4 in the surface of asthmatic ASMCs may be activated by cytokines secreted by the airway epithelial cells and enhance the transmembrane migration of asthmatic ASMCs induced by airway epithelial cells so that it plays a role in airway remodeling of asthma.</p>

Role of toll-like receptor 4 in the asthmatic rat airway smooth muscle cells proliferation and apoptosis / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the role of Toll like receptor 4(TLR4) in the asthmatic rat airway smooth muscle cell (ASMCs) proliferation and apoptosis.</p><p><b>METHODS</b>Established rat model of asthma,isolated and cultured rat ASMCs in asthma, using methods of small molecule RNA interference technology and lipofection method, for small molecule RNA-TLR4 transfection, detected proliferation of ASMCs by MIT minim colorimetry, apoptosis of ASMCs by TUNNEL, the expression of TLR4 protein and mRNA were detected by Western blot and RT-PCR in cells.</p><p><b>RESULTS</b>The proliferation of ASMCs in TNF-alpha group were significantly higher than that in control group and siRNA-TLR4 transfection group and TNF-alpha + siRNA-TLR4 transfection group respectively and the proliferation of ASMCs in siRNA-TLR4 transfction group was lower than that in control group. The apoptosis rate of ASMCs in TNF-alpha group was lower than that in control group, siRNA-TLR4 transfection group and TNF-alpha + siRNA-TLR4 transfection group respectively and the apoptosis rate of ASMCs in siRNA-TLR4 transfection group and TNF-alpha + siRNA-TLR4 transfection group were significantly higher than those in control group. The mRNA and protein expression of TLR4 in control group and TNF-alpha group were significantly higher than those in siRNA-TLR4 transfection group and TNF-alpha + siRNA-TLR4 transfection group. The mRNA and protein expression of TLR4 in TNF-alpha group were significantly higher than those in control group (P < 0.01).</p><p><b>CONCLUSION</b>Activation of TLR4 may contribute to asthmatic airway smooth muscle cell proliferation, inhibiting apoptosis and play an important role in airway remodeling in asthma.</p>

Effect of triptolide on airway smooth muscle proliferation and the expression of nuclear factor-kappa B, Bcl-2 in asthmatic rats / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the effect of triptolide on airway remodeling and the expression of nuclear factor-kappaB, Bcl-2 in asthmatic rats.</p><p><b>METHODS</b>40 rats were randomly divided into 5 groups (n = 8): (1) Control group; (2) Asthmatic 4 week group; (3) Asthmatic 6 week group; (4) Therapeutic 4 week group; (5) Therapeutic 6 week group. The airway resistance and eosinophilic inflammation of airway wall were observed. The airway wall thickness (WA/Pi), the bronchial smooth muscle thickness (smooth muscle area/Pi) and the number of bronchial smooth muscle nucleus (N/Pi) were measured by image analysis system. The expression of PCNA, nuclear factor-kappaB and Bcl-2 protein were determined by immunohistochemical staining and Western blot. The expression of Bcl-2 mRNA was determined by reverse transcription-polymerase chain reaction(RT-PCR).</p><p><b>RESULTS</b>(1) The expression of NF-kappaB protein in asthmatic 4 week group and asthmatic 6 week group was significantly higher than that in control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 week group were significantly lower than those of asthmatic 4 week group, asthmatic 6 week group and therapeutic 4 week group, respectively (P < 0.01, P < 0.01 P < 0.05). (2) The expression of Bcl-2 protein and mRNA of asthmatic 4 week group and asthmatic 6 week group were significantly higher than those in control group respectively (P < 0.01). The expression of Bcl-2 protein of therapeutic 6 week group was significantly lower than those of asthmatic 4 week group, asthmatic 6 week group and therapeutic 4 week group respectively (P < 0.05, P < 0.01, P < 0.01), but the expression of Bcl-2 mRNA was significantly higher than the above-mentioned groups respectively (P < 0.01), the expression of Bcl-2 protein and mRNA of therapeutic 6 week group were higher than control group respectively (P < 0.05, P < 0.01). (3) The expression of PCNA protein of asthmatic 4 week group and asthmatic 6 week group were significantly higher than those of control group respectively (P < 0.01). (4) The WA/ Pi, the smooth muscle area/Pi and the N/Pi of asthmatic 4 week group and asthmatic 6 week group were significantly higher than those of control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 week group were significantly lower than those of asthmatic 4 week group, asthmatic 6 week group and therapeutic 4 week group, respectively (P < 0.01). (5) The airway resistance of asthmatic 4 week group and asthmatic 6 week group were significantly higher than those of the control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 week group were significantly lower than those of asthmatic 4 week group, asthmatic 6 week group and therapeutic 4 week group, respectively (P < 0.01, P < 0.01, P < 0.05).</p><p><b>CONCLUSION</b>The proliferation of airway smooth muscle(ASM) is related with apoptosis of airway smooth muscle cells in asthma. NF-kappaB may be involved in the process. Triptolide may prevent apoptosis of ASMCs and decrease the proliferation of ASM by inhibiting the expression of NF-kappaB, Bcl-2.</p>

Effect of triptolide on airway remodeling and the expression of phosphoinositide 3-kinases in asthmatic rats / 中国应用生理学杂志

<p><b>AIM</b>To explore the effect of Triptolide on airway remodeling and the expression of Phosphoinositide 3-Kinases in asthmatic rats.</p><p><b>METHODS</b>40 rats were randomly divided into 5 groups (n = 8): (1) Control group; (2) Asthmatic 4 weeks group; (3) Asthmatic 6 weeks group; (4) Therapeutic 4 weeks group; (5) Therapeutic 6 weeks group. The airway resistance and eosinophilic inflammation of airway wall were observed. The airway wall thickness (WA/Pi), the bronchial smooth muscle thickness (smooth muscle area/Pi) and the number of bronchial smooth muscle nucleus (N/Pi) were measured by image analysis system. The expression of PI3K protein and mRNA were determined by immunohistochemical staining and reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>(1) The expression of PI3K p85alpha protein and mRNA in asthmatic 4 weeks group and asthmatic 6 weeks group were significantly higher than control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 weeks group were significantly lower than those of asthmatic 4 weeks group, asthmatic 6 weeks group and therapeutic 4 weeks group, respectively (P < 0.01, P < 0.01 P < 0.05). (2) The WA/Pi, the smooth muscle area/Pi and the N/Pi of asthmatic 4 weeks group and asthmatic 6 weeks group were significantly higher than control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 weeks group were significantly lower than those of asthmatic 4 weeks group, asthmatic 6 weeks group and therapeutic 4 weeks group, respectively (P < 0.01). (3) The airway resistance of asthmatic 4 weeks group and asthmatic 6 weeks group were significantly higher than the control group, respectively (P < 0.01). The above-mentioned parameters of therapeutic 6 weeks group were significantly lower than those of asthmatic 4 weeks group, asthmatic 6 weeks group and therapeutic 4 weeks group, respectively (P < 0.01, P < 0.01, P < 0.05).</p><p><b>CONCLUSION</b>The proliferation of airway smooth muscle is a remarkable character of airway remodeling in asthma. The PI3K signal pathway may be involved in the process. Triptolide may reduce AHR and decrease the proliferation of ASMCs by inhibiting the expression of PI3K. It may have potential therapeutic effects in the asthmatic airway remodeling.</p>

Experimental study on the effect of ginkgo bioba extract on the expression of heme oxygenase-1 in bronchial asthma / 中国应用生理学杂志

<p><b>AIM</b>To explore the effect of the ginkgo bioba extract on the expression of heme oxygenase-1 (HO-1) in bronchial asthma.</p><p><b>METHODS</b>30 guinea pigs were randomly divided into 3 groups (n = 10): (1) Normal control group; (2) Asthmatic group; (3) Therapeutic group. Blood carbon monoxide Hb (COHb) percent value, Airway resistance and eosinophilic inflammation of airway wall were observed, the expression of HO-1 in lung tissue were observed by immunohistochemical staining.</p><p><b>RESULTS</b>The expression of HO-1 was mainly located in airway epithelium in these 3 groups, the optical densities were 0.170 +/- 0.020, 0.707 +/- 0.058, 0.397 +/- 0.034, respectively. The asthmatic group showed higher optical densities than that of the normal control group (P < 0.01), and the therapeutic group showed lower optical density than asthmatic group (P < 0.01).</p><p><b>CONCLUSION</b>The expression of HO-1 is inhibited significantly by the treatment of the ginkgo bioba extract, which may be one of the mechanism for treating asthma by ginkgo bioba extract.</p>

The study on expression of heme oxygenase-1 in the peripheral blood mononuclear cell from asthmatic patients / 中国应用生理学杂志

<p><b>AIM</b>To explore the expression of heme oxygenase-1 (HO-1) in the peripheral blood mononuclear cell (PBMC) and relationship to ventilatory function in asthmatic patients.</p><p><b>METHODS</b>Eighteen asthmatic patients and eighteen healthy subjects were selected. HO-1 protein levels in PBMC were measured by immunohistochemical staining and PBMC HO-1 mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR), blood carbon monoxide Hb (COHb) percent value, serum total IgE concentration and pulmonary ventilatory function were observed in asthmatic patients and healthy subjects.</p><p><b>RESULTS</b>The percentage of cells in immunohistochemical staining positive staining of HO-1 were significantly higher in asthmatic patients (41.7 +/- 7.44%) compared with that of healthy subjects (10.5 +/- 4.36%, P < 0.01), the optical densities of PBMC HO-1 mRNA were higher in asthmatic patients (26.05 +/- 4.14) compared with that of healthy subjects (10.82 +/- 4.26, P < 0.01). The relation analysis showed PBMC HO-1 protein levels had significantly negative relation with FEV1, PEFR, MEFR50%, respectively (r = -0.89, -0.56, -0.51, P < 0.01, respectively) and positive relation with COHb percent value, serum total I gE concentration (r = 0.80, 0.48, P < 0.05, respectively), and PBMC HO-1 mRNA levels had significantly negative relation with FEV1, PEFR, MEFR50%, respectively (r = -0.89, -0.65, -0.67, P < 0.05, respectively) and positive relation with COHb percent value, serum total IgE concentration (r = 0.85, 0.62, P < 0.01, respectively).</p><p><b>CONCLUSION</b>The expression of PBMC HO-1 protein and mRNA are increased significantly in asthmatic patients, HO-1 may play a significant role in the pathogenesis of asthma. The expression of HO-1 has relation with severity of asthma.</p>