[Effects of medicinal extract for tonifying kidney to relieve asthma on glucocorticoid receptor expression in lung tissues of rats with bronchial asthma].

OBJECTIVE: To observe the effects of medicinal extract for tonifying kidney to relieve asthma on glucocorticoid receptor (GR) expression in rats with asthma, and to explore its mechanism in treating asthma. METHODS: Sixty SD rats were randomly divided into normal control group, untreated group, dexamethasone group, and medicinal extract-prevented, medicinal extract-treated, and medicinal extract-prevented and -treated groups, with ten rats in each group. Asthma was induced by intraperitoneal injection of ovalbumin (OVA) and forced inhalation of atomized OVA. Expression of GR in lung tissues was detected by immunohistochemical method. Pathological changes of the lung tissues were observed by HE straining. RESULTS: Expression of GR was lower in the untreated group than in the normal control group (P<0.05). Expressions of GR in medicinal extract groups were up-regulated as compared with those in the untreated group and dexamethasone group (P<0.05, P<0.01), and there were no significant differences as compared with the normal control group (P>0.05). CONCLUSION: Medicinal extract for tonifying kidney to relieve asthma can increase the expression of GR in lung tissues of asthmatic rats, which may be one of its mechanisms in preventing and treating asthma.

[Effects of Kechuanluo oral liquid on eosinophil apoptosis and its regulation factors in lung tissues of asthmatic mice].

OBJECTIVE: To explore the mechanism of Kechuanluo oral liquid, a compound Chinese herbal medicine, in inhibiting allergic airway inflammation by observing the effects of Kechuanluo on eosinophil (EOS) apoptosis and its regulation factors in asthmatic mice. METHODS: Fifty-six BALB/c mice were randomly divided into normal control group (n=16), untreated group (n=16), Western medicine group (n=12) and Kechuanluo group (n=12). Except for the mice in normal control group, asthma was induced in BALB/c mice by using ovalbumin (OVA) and potassium aluminium sulfate. The mice were intragastrically administered with normal saline, Kechuanluo (30 ml/kg daily) and prednisolone tablets (10 mg/kg daily) respectively for two weeks. At 0 hour, the 3rd, 7th and 14th day after the end of OVA sensitization, the EOSs of lung tissues were counted by improved-Giemsa staining method; immunohistochemical method and image analysis were used to detect the expressions of Fas, FasL and Bcl-XL in the EOSs in the four groups; and apoptotic rates of the EOSs in the lung tissues of asthmatic mice were detected by terminal deoxynucleotidyl transferase-mediated biotin-dUTP nick end labeling (TUNEL) technique. RESULTS: Compared with the untreated group, airway inflammations of the mice in the Kechualuo group and Western medicine group were lessened and the EOS counts decreased on the 3rd, 7th and 14th day after the last OVA sensitization (P<0.05). On the 3rd day, the EOSs apoptotic rates, the expression areas of Fas in the EOSs and FasL in the lung tissues were significantly higher in the Kechuanluo group than those in the untreated group (P<0.01), furthermore, the EOS apoptotic rate reached the peak level. Inversely, the EOS count and the expression area of Bcl-XL in the EOSs were obviously lower in the Kechuanluo group (P<0.05). On the 7th and 14th day, the expression areas of Fas and Bcl-XL in the EOSs were significantly decreased in the Kechuanluo group (P<0.01), and on the 14th day, the EOS apoptotic rate and the expression area of FasL in the lung tissues were obviously lower either. The effects exhibited in the Western medicine group were similar to those in the Kechuanluo group. CONCLUSION: There is airway inflammation with eosinophilic infiltration in asthmatic mice, accompanied with suppressed apoptosis and delayed apoptosis. Kechuanluo can induce and accelerate EOS apoptosis in early inflammation by inhibiting eosinophilic inflammation, improving Fas expression in EOSs and FasL expression in the lung tissues, and reducing Bcl-XL expression in EOSs.

[Function of MMP/TIMP on airway remodeling of bronchial asthma and treating effects of Zhichuan capsule].

OBJECTIVE: To explore the mechanism of the bronchial asthma and to study the treating effects of Zhichuan Capsule on the airway remodeling of asthmatic model rats. METHODS: The rat model was established by being sensitized and activated with different density of ovalbumin through prolonged and repeated exposure for 8 weeks. The rats were randomly divided into model group, Zhichuan Capsule treated group, dexameson treated group, and Zhichuan Capsule and dexameson treated group. Another group of normal rats were taken as control. General histological changes were observed by hematoxylin and eosin stained sections. Being standardized by internal perimeter (Pi), the wall thickness (d), internal area (Ai), outer area (Ao) and wall area (WA) of the airway were quantified by computer-assisted image analysis system. The express of MMP-9, TIMP-1, Col I, Col III and ColV in the airway were examined by immunocytochemical methods. During the course of airway remodeling, the dynamic changes of model rats were observed at different time points (2, 4, 6 and 8 weeks after the activating). Statistical comparison was performed by ANOVA followed by Fisher LSD test. RESULTS: (1) Histologic examination showed eosinophil infiltration within the airway walls, epithelial damage, excessive mucus in the lumen and edema in the submucosa of the airways in model rats, and that the collagen deposition increased accompanied by increasing of TIMP-1. In the model rats, MMP-9 increased at the time point of 2 weeks, but it decreased in the late stage (8 weeks after activating) of airway remodeling. And the level of TIMP-1 was far higher than MMP-9 at the time point of 8 weeks. (2) Zhichuan Capsule could down-regulate the level of TIMP-1 in the airway wall, as well as the thickness of airway wall and the collagen deposition. And there were progressing effects when it was used together with dexameson. CONCLUSION: (1) The early increase of MMP-9 is a key point to start remodeling; and the increase of TIMP-1 in the late stage, which inhibits collagenase activity, may play an important role in developing airway fibrosis. Imbalance between MMP-9 and TIMP-1 is a marker of airway remodeling. (2) Zhichuan Capsule can decrease the deposition of collagen and suppress the airway remodeling by inhibiting the TIMP-1 expression.

[Morphological changes following airway remodeling in asthmatic rats].

OBJECTIVE: To explore the establishment of an asthmatic model with airway remodeling in rats by observing the morphological changes of the airway in different stages. METHODS: Animals were divided into two groups: asthmatic group and control group. The subjects were observed at 5 phases: before activation and 2, 4, 6, 8 weeks after activation. General histological changes were observed using hematoxylin and eosin stained sections. Being standardized by internal perimeter (Pi), the wall thickness (d), internal area (Ai), outer area (Ao) and wall area (WA) of the airway were quantified by computer-assisted image analysis system. And the membrane airways were divided into 3 grades (large, media and small airways) to be analyzed. RESULTS: The changes of small airway appeared earlier and greater than the others. The changes of media airway appeared later than the small one. The changes of large airway appeared as later as the 4th week after activation. There were no differences before and after activation in control group (P>0.05). CONCLUSION: (1) Prolonged, repeated and low density Ag activation could establish asthmatic model with airway remodeling. (2) The morphological changes of the airways increased following airway remodeling, and the greatest increase occurred at small airway. (3)The morphological changes in the model rats were similar to that in human beings.

GM-CSF and IFN-gamma-induced expression of human leucocyte antigen class II molecules on basophils of umbilical cord blood.

AIM: To determine whether basophils expressed human leucocyte antigen (HLA) class II molecules. METHODS: Basophils in umbilical cord blood were separated and purified with methods of density gradient centrifugation and immunomagnetic microbeads. The isolated basophils were cultured in RPMI-1640 plus 10 % fetal calf serum at 37 in a humidified atmosphere with 5 % CO2 and stimulated with granulocyte-macrophage colony-stimulating factor (GM-CSF) or interferon (IFN)-gamma for 20-60 h. The expression of HLA class II molecules on basophils was measured with fluorescence activated cell sorter (FACS). RESULTS: The purity of isolated basophils was > or = 83.5 %. After treated with GM-CSF 10 microg/L or IFN-gamma 100 kU/L, the expression of HLA class II molecules became detectable on membranous surface and at a higher level at 20 h, the percentage of expression was 10.2 %+/-2.1 % and 11.3 %+/-1.0 %, respectively. CONCLUSION: Basophils possessed the potency of HLA class II molecular expression.