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Objective:To investigate the effects of nebulization with high-dose budesonide (BUD) combined with ipratropium bromide (IB) on airway remodeling and mucus secretion in patients with acute exacerbations of chronic obstructive pulmonary disease (AECOPD).Methods:Ninety patients with AECOPD who received treatment in Wenling Hospital of Traditional Chinese Medicine between January 2020 and June 2021 were included in this study. They were assigned to the conventional-dose group ( n = 45, odd number) and high-dose group ( n = 45, even number) according to the number of admission. In the conventional-dose group, nebulization with IB (0.5 mg administered within 20 minutes, three times daily) and BUD (2 mg administered within 20 minutes, twice daily) was performed. In the high-dose group, nebulization with IB (0.5 mg administered within 20 minutes, three times daily) and BUD (4 mg administered within 20 minutes, twice daily) was performed. After nebulization, a mouthwash was required in each group. After 7 days of treatment, clinical efficacy was compared between the two groups. Before and 7 days after treatment, airway remodeling level (matrix metalloproteinase-9 and interleukin-8), airway mucus secretion (mucin-5ac and neutrophil elastase) and the incidence of adverse reactions were compared between the two groups. Results:Total response rate in the high-dose group was significantly higher than that in the conventional-dose group [95.56% (43/45) vs. 82.22% (37/45), χ2 = 4.05, P < 0.05]. After 7 days of treatment, serum matrix metalloproteinase-9 and interleukin-8 levels in the high-dose group were (416.96 ± 30.96) μg/L and (6.33 ± 1.03) μg/L, respectively, which were significantly lower than those in the conventional-dose group [(452.25 ± 32.16) μg/L, (7.85 ± 1.24) μg/L, t = 5.30, 6.32, both P < 0.001)]. After 7 days of treatment, serum mucin-5ac and neutrophil elastase levels in the high-dose group were (1.33 ± 0.21) μg/L and (4.06 ± 1.03) μg/L, respectively, which were significantly lower than those in the conventional-dose group [(1.58 ± 0.23) μg/L, (5.11 ± 1.14) μg/L, t = 5.38, 4.58, both P < 0.001]. There was no significant difference in incidence of adverse reactions between high-dose and conventional-dose groups [8.89% (4/45) vs. 4.44% (2/45), χ2 = 0.71, P > 0.05). Conclusion:Nebulization with high-dose BUD combined with IB for treatment of AECOPD can improve airway remodeling, reduce airway mucus hypersecretion and has definite therapeutic effects. Findings from this study are of great innovation and science.
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Objective:To investigate the changes of gene or protein expression and activity of matrix metalloprotein9 (MMP9) and tissue inhibitor ofmetalloproteinasel (TIMP1) in the carotid artery (CA) of 4 wk hindlimb unweighted rat.Methods:A 4 weeks(wk) hindlimb unweighted (HU) rat model was used to simulate the effect of weightlessness on the cardiovascular system.Transmission electron microscopy was used to detect the content of ECM.Reverse transcriptase polymerase chain reaction(RT-PCR) was conducted to measure the mRNA content MMP and TIMP1.Immunohistochemistry and Western blot technique were used to measure the protein abundance.Gelatin zymography was carried out to detect the activity of MMP9.Results:Compared to the control group (CON),the area of ECM was enhanced (P＜0.05) and the content of collage fiber was increased (P＜0.05) in the CA of HU rats;moreover,HU did not affect the mRNA expression of MMP9,but significantly reduced the protein content (P＜0.05) or enzymatic activity (P＜0.05).Accordingly,the mRNA or protein expression of TIMP1 in the CA was significantly increased by HU (P＜0.05).Conclusion:Simulated weightlessness caused imbalance between MMP and TIMP1 expression,which might contribute to the ECM aggregation and stiffness of CA.
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Objective To discuss the role of p38MAPK signal pathway in the process of TWEAK inducing rheumatoid arthritis(RA) fibroblast-like synoviocyte(FLS) to synthesize MMP-9 and look for a new target for RA treatment. Methods RA FLS were primarily cultured and stimulated with TWEAK. Western blot was used to detect the expression of p-p38MAPK and p65 in RA FLS. FLS were pretreated by SB203580 or not. ELISA was used to detect the concentration of MMP-9 in cell-cultured fluid. The mRNA expression of MMP-9 was measured by RT-PCR. Results TWEAK( 100 ng/ml) can make p38MAPK phosphorylated and increased the expression of p65 protein in the cell nucleus. SB203580 can partially inhibit the expression of MMP-9 and MMP-9 mRNA produced by RA FLS which is induced by TWEAK. Conclusion TWEAK induced RA FLS to synthesize MMP-9, in that process, the p38MAPK signal trausduction pathway was in active state, and induced the expression of NF-κB.
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Objective To construct recombinant adeno-associated virus and lentivirus carrying siRNA of TIMP-1 and to investigate the efficiency of infection and short-term inhibitory effect of TIMP-1 gene expres-sion on rat hepatic stellate cells. Methods One pair of siRNA which could effectively inhibit expression of the TIMP-1 gene in HSC-T6 was screened and cloned into AAV vector and lentiviral vector to construct the recombinant AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP. AAV/GFP and Lenti/GFP as neg-ative control were also obtained. Experiments were assigned to five groups: AAV/siRNA-TIMP-1/GFP, AAV/GFP, Lenti/siRNA-TIMP-1/GFP, Lenti/GFP group and mock group. Rat HSC-T6 cells were infected by these recombinant viruses at a concentration of MOI by 10. To monitor the efficiency of infection, fluores-cence microscope and flow cytometer were used. After 7 d post-infection, Western blot was used to detect the TIMP-1 protein expression. Results HSC-T6 had no significant changes after infection. The efficiency of infection in AAV/GFP and Lenti/GFP group were 72.7% and 70.0%, AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP group were 64.58% and 61.86%. The protein expression levels of TIMP-1 in HSC-T6 cells at 7 d post-infection by the recombinant AAV and Lentivirus were decreased 40.0% compared with those in mock control and normal HSC-T6 (P<0.05). Conclusion Recombinant AAV/siRNA-TIMP-1/GFP and Lenti/siRNA-TIMP-1/GFP could effectively infect HSC-T6 with similar efficiency and suppress the expression of TIMP-1 in rat HSC-T6 remarkably.
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Objective This study was to investigate the role of STAT3 signaling pathway in gastric cancer invasiveness. Methods Janus kinase 2 selective inhibitor AG490 was used to inhibit the activation of STAT3 signaling pathway. Matrigel-coated transwell was employed to estimate the invasiveness of BGG-823 cells. Alterations in the activity of STAT3 and expression of MMPs and TIMPs were measured by Western blot and RT-PCR. Results The activation of STAT3 signaling pathway was suppressed by AG490. AC490 significantly decreased the invasiveness of gastric cancer cell line BGC-823 through Matrigel-coated filters. Decreased invasion was associated with the inhibition of MMP-2, and MMP-9 at both mRNA and protein levels by RT-PCR and Western blot. Meanwhile, the expression of TIMP-1, and TIMP-2 were up-regulated at mRNA levels. Conclusion Inhibition of STAT3 signaling pathway in AG490 suppresses gastric cancer cell invasiveness through down-regulation of several invasion-related factors including MMP-2, and MMP-9 and up-regulation of their inhibitors.
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Objective To detect both the protein and gene expression of matrix metalloproteinase (MMP 2,MMP 9) and its tissue inhibitor (TIMP 2) in tissues of bladder transitional cell carcinoma (BTCC), and to study the correlation with the grading and staging of the neoplasm and with the prognosis. Methods 41 human BTCC ,15 normal bladder tissue were studied by Western Blot and RT PCR analysis followed by computer assisted image analysis in order to detect the A values of their expression. Results In BTCC, the A value of MMP 2, MMP 9 were increased significantly as compared to normal bladder epithelium. There was no statistical significant difference of A value of TIMP 2 between normal bladder tissue and bladder cancer tissue. The ratio of MMP 2/TIMP 2 in bladder cancer showed statistical significantly difference as compared with normal bladder tissue. Conclusions This study demonstrated there was a high expression of MMP 2 and MMP 9 in BTCC,by which collagen Ⅳ inside basement membrane of bladder was damaged.However, the ratio of MMP 2/TIMP 2 has more prognostic value than the expression of MMP 2 and MMP 9 alone.
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Objective To study the effect of Kangxian Ruangan Granule(KRG)on the collagen degradation in hepatic astrocytes(HA)at the cellular and molecular level.Methods The medicated sera of KRG was used to i ncubate the sub-cultured HA and sodium chloride and c olchicines were taken as controls.T he content of typeⅠcollagen in the cultured su-pernatant was measured with ELISA an d then was corrected by assaying the t otal protein in cell layer.The mRNA e xpres-sions of matrix metalloproteinase -1(MMP1)and tissue inhibitor of metalloproteinase -1(TIMP -1)were analyzed by semiquantitative RT -PCR.Results The content of typeⅠcollagen in the KRG -cultured supern atant was lower than that in the other two groups(P
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Cardiotoxin II of the Indian cobra (Naja naja) contains approximately four Mg2+ per mol. Complete demetallation of the toxin is achieved by three cycles of treatment with ethylenediamine tetraacetate and gel filtration. Reconstitution of toxin by treatment of the apo-protein with Mg2+ restores metal content and inorganic pyrophosphatase activity only to the extent of two atoms/mol and 65%, respectively. Use of Mg (II)-EDTA in the reconstitution experiment yields restoration of half the original enzyme activity. Mg2+ is required for the inorganic pyrophosphatase action of the toxin. A definitive statement on the non-essentiality of Mg2+ for the lethal toxicity of the toxin is not possible at present, although experimental observations indicate that demetallated toxin is as toxic as the native toxin. Based on this and the differing sensitivities of the enzyme and toxic activities of the toxin to heat, it is suggested that the reaction centres in the toxin for the two activities are different and that the pyrophosphatase activity is not causally connected with the lethal toxicity of the toxin.