RESUMO
BACKGROUND: It is generally thought that the pathogenesis of the chronic blocking pulmonary disease (COPD) is characteristics in chronic inflammations of air passage, lung essence and the lung blood vessels. As the classic Chinese medicine, Maxing Shigan decoction has the pharmaco logical effects on fever alleviation, anti-inflammation, anti-virus, cough relieving and phlegm resolving, which is suitable for the treatment of phlegm heat cumber lung COPD, and is remarkable in the curative effect.OBJECTIVE: To probe into the mechanism of Maxing Shigan decoction ini the treatment of COPD by studying the changes in interleukin-8 (IL-8),tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-4 (IL-4)and the ratio between IFN-γ and IL-4 in rats with COPD.DESIGN: Randomized control animal experiment.SETTING: Affiliated Hospital of Shandong University of Traditional Chinese Medicine.MATERIALS: Seventy-two healthy male Wistar rats of SPF level with the body mass of (200±20) g were selected. Maxing Shigan decoction, composed of 5 g of ephedra, 9 g of almond, 6 g of liquorices, 18 g of gypsum.The water-decocted enrichment was 0.475 g/mL, and the medical herb pieces were supplied by the TCM Dispensary of Affiliated Hospital of Shandong Medical University of Traditional Chinese Medicine.METHODS: The experiment was conducted in the Central Laboratory,Affiliated Hospital of Shandong University of Traditional Chinese Medicine from April to October 2005. ① 72 healthy male Wistar rats were randomly divided into 3 groups: blank control group (n=24), retention of phlegm-heat modeling group (modeling group, n=24) and retention of phlegm-heat treatment group (treatment group, n=24). Improved smoking and trachea dropping lipopolysaccharide was adopted to copy COPD model, and model of phlegm heat cumber lung was remodeled for copy.② Rats in the treatment group were lavaged with 2 mL of Maxing Shigan decoction per day (the medicine composition: Chinese ephedra 5 g, almond 9 g, licorice 6 g, gypsum 18 g, the water fries the concentration for the density 475 g/L concentrated solution) from the 30th day, while rats in the modeling group were lavaged with 2 mL of normal saline each day.Rats in the blank control group received no interventions, which ate and drank freely under the same environment. ③ The contents of IL-8, IL-4 and TNF-α in the homogenate of lung tissues were tested by radio-immune method. ④ The content of IFN-γ in the homogenate of lung tissues was detected by the enzyme immunoassay method. ⑤ The ratio of IFN-γ and IL-4 was calculated. MAIN OUTCOME MEASURES: The contents of IL-8, TNF-α, IFN-γ,IL-4 as well as the ratio between IFN-γ and IL-4 in the homogenate of lung tissues in rats of each group.RESULTS: ① The level of IL-8 in the modeling group was markedly higher than that in the blank control group [(581.41±74.72), (318.63±40.01 ) ng/L,P ＜ 0.01], which was significantly decreased after the treatment [(533.95±68.67) ng/L,P ＜ 0.01], while there was difference in comparison with the blank control group (P ＜ 0.05). ② The level of TNF-α in the modeling group was significantly higher than that in the blank control group [(160.78±20.90), (100.29±24.2) ng/L, P ＜ 0.01], which was remarkably decreased after the treatment [(147.08±19.43) ng/L,P ＜ 0.05],whereas it was different in comparison with that in the blank control group(P ＜ 0.01). ③ Compared with the blank control group, the level of IFN-γ in the modeling group was markedly decreased [(230.32±72.33), (328.08±50.57) ng/L ,P＜ 0.01], which was obviously increased after the treatment [(267.24±55.14) ng/L ,P ＜ 0.05], and there were differences between the two groups (P ＜ 0.01). ④ The level of IL-4 was significantly increased in the modeling group than that in the blank control group[(135.08±29.17), (82.41±14.66) ng/L,P ＜ 0.01], which was significantly decreased after the treatment [(107.99±23.76) ng/L,P ＜ 0.01], and there was difference between the two groups (P ＜ 0.01 ). ⑤ The ratio between IFN-γ and IL-4 declined markedly in the modeling group than that in the blank control group (1.77±0.73,4.09±0.91 ,P ＜ 0.01), and the ratio increased markedly after the treatment(2.59±0.79, P ＜ 0.01 ). There was difference between the two groups(P ＜ 0.01 ).CONCLUSION: Maxing Shigan decoction can regulate the changes in level of multiple cytokines in rats with COPD, which has the effects of anti-inflammation and enhancing the immunity of organism immunization with clear curative effect on COPD (syndrome of retention of phlegmheat).
RESUMO
BACKGROUND: Many diseases accompany with changes of gene expression which can provide a valuable clue for pathogenesis and therapy of cognitive diseases.OBJECTIVE: To screen expression of different genes in lung tissue of model rats with chronic obstructive pulmonary disease (COPD) by gene expression profiling technique. DESIGN: Open study. SETTING: Affiliated Hospital of Shandong University of Traditional Chinese Medicine; Radiation Medical Institute of Academy of Military Medical Sciences of Chinese PLA; Shangdong University of Traditional Chinese Medicine.MATERIALS: The experiment was carried out in the Central Laboratory of Affiliated Hospital, Shandong University of Traditional Chinese Medicine from April to October 2005. ① Fifteen healthy male Wistar rats of SPF grade and aged 50 days were randomly divided into normal group, model control group and Chinese herb group with 5 in each group. ② Type of gene chip was BiostarR-40S and provided by Shanghai Boxing Gene Chip Company Limited (batch number: G050510010052).METHODS: ① COPD models in model control group and Chinese herb group were established with modified smoking-fumigated method plus adding lipopolysaccharide in windpipe. Pathological changes of airway were observed under optic microscope. Models were established successfully based on the following phenomenon: hyperemia and edema in mucous membrane of airway; degeneration and necrosis of epithelial cells; infiltration of bronchial cavity in lung tissue and surrounding chronic inflammatory cells; proliferation of smooth muscle and fibrocyte surrounding dissepiment; thin and broken interval; amplifying confluence of pulmonary alveolus; thickness of vascular wall of arteriole; decrease of vaso-cavity; infiltration of surrounding inflammatory cells. ② At 30 days after modeling,0.65 g/mL self-made Chinese herb (mahuang, xingren, huangqi, etc.) wasperfused into rats in Chinese herb group with 2 mL each time, once a day,for successive 14 days. Rats in model control group were perfused with 2 mLsaline; however, rats in normal group were untouched. All rats drank freely under the same internal environment. ③ After administration, rats in each group were sacrificed to extract total RNA, label and hybridizated with probe, wash pieces, analyze gray value of clip with clip imaging software,obtain original signal of each gene point (signal values of foreground and background) and determine differently expressed genes. If size of test was more than 2.0, genes were regarded as up-regulation genes; otherwise, if ratio was less than 0.5, they were regarded as down-regulation genes. In this study, based on reliability, if size of test was more than 2.5, they were regarded as up-relation genes; however, if ratio was less than 0.375, they were regarded as down-regulation genes.MAIN OUTCOME MEASURES: ① Comparisons of differently expressed genes in normal and model control groups; ② Comparisons of differently expressed genes in normal and Chinese herb groups; ③Comparisons of differently expressed genes in model control and Chinese herb groups.RESULTS: Fifteen rats were all involved in the final analysis. ① As compared with those in normal group, there were 57 differently expressed genes in model control group, involving in immunity, metabolism, signal transduction, adjusting of gene expression, cell cycle, cell transport, cell migration, fibrosis, etc. ② Gene expression in Chinese herb group reached normal level and there were 11 differently expressed genes as compared with those in normal group, involving in stress, signal transduction, cytoskeleton, etc. ③ As compared with those in model control group, there were 7 differently expressed genes in Chinese herb group, involving in immunity and excretion of neurotransmitter.CONCLUSION: Changes of gene expression may be one of reasons of COPD pathogeneses; moreover, most differently expressed genes can recover the normal level after drug therapy which is beneficial to correct pathological changes of COPD.