BACKGROUNDSo far, there is no efficient treatment for pulmonary fibrosis. The objective of this study was to determine whether intramuscular injection of the hepatocyte growth factor (HGF) plasmid DNA by in vivo electroporation could prevent bleomycin-induced pulmonary fibrosis in rats, and to investigate the possible mechanisms.

METHODSTwenty male Wistar rats were randomly divided into four groups: control group (group C), model group (group M), early intervention group (group I) and late intervention group (group II). Groups M, I and II were intratracheally infused with bleomycin, then injected the plasmid pcDNA3.1-hHGF to group I on day 7, 14 and 21. Group II received the same treatment like Group I on day 14 and 21. All the rats were killed on day 28 after bleomycin injection. We detected Homo HGF expression in the rats with ELISA method and estimated the pathological fibrosis score of lung tissue using hematoxylin eosin (HE) and Massion staining. The mRNA expression of transforming growth factor-beta1 (TGF-beta1), cycloxygenase-2 (COX-2), and rat HGF in rat pulmonary parenchyma were evaluated by RT-PCR. Immunohistochemistry and Western blotting were performed to determine the protein expression of transforming TGF-beta1 and COX-2 in lung parenchyma.

RESULTSThe plasmid pcDNA3.1-hHGF could express hHGF in NIH3T3 cells and the hHGF protein is secreted into the culture medium. The expression of hHGF protein could be monitored in quadriceps muscle, plasma and lung in Groups I and II. Pulmonary fibrosis levels of Groups I and II were obviously lower than that of group M (P < 0.05). Expression of TGF-beta1 protein and mRNA in lung tissue was markedly decreased in Groups I and II compared with Group M (P < 0.05). The level of expression of HGF and COX-2 mRNA was higher in Groups I and II than in Group M (P < 0.05).

CONCLUSIONSInjection of the plasmid pcDNA3.1-hHGF into skeletal muscle with electroporation has a potential role in the treatment of bleomycin-induced lung fibrosis. Exogenous HGF may inhibit the expression of TGF-beta1 and regulate the crosstalk between AECs and mesenchymal fibroblasts.

Animals ; Bleomycin ; toxicity ; Cyclooxygenase 2 ; genetics ; Electroporation ; Genetic Therapy ; Hepatocyte Growth Factor ; genetics ; Hydroxyproline ; analysis ; Injections, Intramuscular ; Lung ; chemistry ; Male ; Muscle, Skeletal ; metabolism ; Plasmids ; Pulmonary Fibrosis ; chemically induced ; therapy ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Transforming Growth Factor beta1 ; analysis ; genetics

To optimize the belt drying process conditions optimization of Gardeniae Fructus extract from Reduning injection by Box-Behnken design-response surface methodology, on the basis of single factor experiment, a three-factor and three-level Box-Behnken experimental design was employed to optimize the drying technology of Gardeniae Fructus extract from Reduning injection. With drying temperature, drying time, feeding speed as independent variables and the content of geniposide as dependent variable, the experimental data were fitted to a second order polynomial equation, establishing the mathematical relationship between the content of geniposide and respective variables. With the experimental data analyzed by Design-Expert 8. 0. 6, the optimal drying parameter was as follows: the drying temperature was 98.5 degrees C , the drying time was 89 min, the feeding speed was 99.8 r x min(-1). Three verification experiments were taked under this technology and the measured average content of geniposide was 564. 108 mg x g(-1), which was close to the model prediction: 563. 307 mg x g(-1). According to the verification test, the Gardeniae Fructus belt drying process is steady and feasible. So single factor experiments combined with response surface method (RSM) could be used to optimize the drying technology of Reduning injection Gardenia extract.
Chemistry, Pharmaceutical ; instrumentation ; methods ; Desiccation ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Gardenia ; chemistry ; Research Design ; Vacuum

OBJECTIVETo investigate the molecular mechanism of inhibitory effect of Salvia miltiorrhiza Injection (SMI) coordinated with dexamethasone (DXM) on allergic airway inflammation in asthmatic rats.

METHODSForty SD rats were randomly divided into 5 groups equally: the normal group, the asthma model group, the DXM group, the SMI group and the DXM + SMI group, they were treated with correspondant herbal medicines. Pathologic changes of lung tissue were obseved with HE stain, count of WBC and eosinophil (Eos) in bronchoalveolar lavage fluid (BALF) were estimated and the expressions of interleukin-13 (IL-13) and Eotaxin in lung tissue were measured by RT-PCR and SP method of immunohistochemistry assay.

RESULTSThere was moderate inflammation in lung tissue in the SMI group, and mild inflammation in the DXM + SMI and the DXM group, which was similar to that in the normal group. Compared with the asthma model group, Eos and WBC count in BALF and the expression of IL-13 and Eotaxin in the lung tissue were significantly lower in the three treated groups (P < 0.05), particularly in the DXM + SMI group, showing a significant difference as compared with the other two groups (P < 0.05 or P < 0.01). Additionally, IL-13 expression was positively correlated with Eotaxin expression (r = 0.92, P < 0.01).

CONCLUSIONSMI could inhibit the expression of IL-13 and Eotaxin in the lung of asthmatic rats, showing inhibitory effects synergistic with DXM on airway inflammation.

Animals ; Anti-Inflammatory Agents ; administration & dosage ; pharmacology ; Asthma ; drug therapy ; genetics ; metabolism ; Chemokine CCL11 ; biosynthesis ; genetics ; Dexamethasone ; administration & dosage ; pharmacology ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Eosinophils ; drug effects ; metabolism ; Immunohistochemistry ; Injections, Intraperitoneal ; Interleukin-13 ; biosynthesis ; genetics ; Lung ; drug effects ; metabolism ; pathology ; Male ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo explore the effect of Shenmai Injection (SMI) on L-type calcium channel of diaphragmatic muscle cells in rats.

METHODSSingle diaphragmatic muscle cell of rats was obtained by the acute enzyme isolation method and the standard whole-cell patch clamp technique was used to record the inward peak L-type calcium current (IPLC) and current-voltage relationship curve of diaphragmatic muscle cells of 7 rats, and to compare the effects of SMI in various concentrations on them.

RESULTSWhen keeping the electric potential at -80 mV, stimulation frequency 0.5 Hz, clamp time 300 ms, stepped voltage 10 mV, and depolarized to +60 mV, 10 microliters/ml of SMI could only cause the mean IPLC of rat's diaphragmatic muscle cells increased from -6.9 +/- 0.6 pA/pF to -7.5 +/- 0.7 pA/pF, the amplification being (9.2 +/- 2.8)%, comparison between those of pre-treatment and post-treatment showed insignificant difference. But when the concentration of SMI increased to 50 microliters/ml and 100 microliters/ml, the mean IPLC increased to -8.4 +/- 0.6 pA/pF and -9.2 +/- 0.6 pA/pF, respectively, and the amplification was (22.4 +/- 1.7)% and (34.6 +/- 4.6)% respectively, showing significant difference to that of pre-treatment (P < 0.05). However, SMI showed no significant effect on maximal activation potential and reversal potential.

CONCLUSIONSMI can activate the calcium channel of diaphragmatic muscle cells in rats, increase the influx of Ca2+, so as to strengthen the contraction of diaphragmatic muscle, which may be one of the ionic channel mechanisms of SMI in treating diaphragmatic muscle fatigue in clinical practice.

Animals ; Calcium Channels, L-Type ; metabolism ; Diaphragm ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; Female ; Male ; Muscle Contraction ; drug effects ; Muscle Fibers, Skeletal ; metabolism ; Patch-Clamp Techniques ; Plant Extracts ; pharmacology ; Rats ; Rats, Wistar

To investigate the effects of matrine on apoptosis and expression of adhesion molecules in human multiple myeloma cell line RPMI8226 cells, RPMI8226 cells were incubated with indicated concentrations of matrine. The growth of RPMI8226 cells was observed by CCK-8 colorimetric assay and apoptosis was detected by flow cytometry using Annexin V-FITC/PI staining. The cell cycles were analyzed by PI staining. Flow cytometry using Annexin V-FITC/PI staining was used to detect the expression of cell adhesion molecules, including CD44, CD44v6, CD54 and CD106. The results showed that RPMI8226 cell viability in presence of matrine decreased markedly in a dose- and time-dependent manners. The apoptosis could be induced by matrine and its level increased following the augmentation of the drug concentration. After treated by matrine for 48 hours, a concentration-dependent increase of cells in G(0)/G(1) phase and a decrease in S phase could be detected, but no obvious change of cell count was found in G(2)/M phase. Treatment of RPMI8226 cells with matrine for 48 hours resulted in decrease of expression levels of CD44 and CD54, while expressions of CD44v6 and CD106 had no significant change. It is concluded that matrine induces in vitro apoptosis, suppresses proliferation in multiple myeloma cells and depresses expression of some adhesion molecules.
Alkaloids ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Hyaluronan Receptors ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Multiple Myeloma ; pathology ; Quinolizines ; pharmacology

BACKGROUNDThe decrease of surfactant protein (SP) secreted by the alveolar type II cell is one of the important causes of limiting air of pulmonary emphysema. However, the SP-A gene and protein changes in this disease are rarely studied. This study was undertaken to investigate alterations in SP-A gene activity and protein, and to explore their roles in the pathogenesis of emphysematous changes.

METHODSTwenty Wistar rats were divided randomly into a normal control group (n = 10) and a cigarette smoking (CS) + lipopolysaccharide (LPS) group (n = 10). Ultra-structural changes were observed under an electron microscope. The number of cells positive for SP-A was measured by immunohistochemistry. The mRNA expression and protein level of SP-A in the lung tissues were determined by quantitative polymerase chain reaction (qPCR) and Western blot separately. The protein level of SP-A in lavage fluid was determined by Western blot.

RESULTSThe number of cells positive for SP-A of the CS + LPS group (0.35 +/- 0.03) was lower than that of the blank control group (0.72 +/- 0.06, P < 0.05). The level of SP-A in the lung tissues of rats in the CS + LPS group (0.2765 +/- 0.0890) was lower than that in the blank control group (0.6875 +/- 0.1578, P < 0.05). The level of SP-A in the lavage fluid of rats in the CS + LPS group (0.8567 +/- 0.1458) was lower than that in the blank control group (1.3541 +/- 0.2475, P < 0.05). The lung tissues of rats in the CS + LPS group showed an approximate increase (0.4-fold) in SP-A mRNA levels relative to beta-actin mRNA (P < 0.05).

CONCLUSIONSThe changes of SP-A may be related to emphysematous changes in the lung. And cigarette smoke and LPS alter lung SP-A gene activity and protein homeostasis.

Animals ; Blotting, Western ; Emphysema ; metabolism ; pathology ; Homeostasis ; Immunohistochemistry ; Male ; Microscopy, Electron ; Polymerase Chain Reaction ; Pulmonary Surfactant-Associated Protein A ; analysis ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar

OBJECTIVETo explore the protective effect of tanshinone II A on lipopolysaccharide (LPS)-induced lung injury in rats, and possible mechanism.

METHODSLPS (O(111): B4) was used to produce a rat model of acute lung injury. Sprague-Dawley rats were randomly divided into 3 groups (8 in each group): the control group, the model group (ALI group), and the tanshinone II A treatment group. Expression of adhesion molecule CD18 on the surface of polymorphonuclear neutrophil (PMNCD18) in venous white blood cells (WBC), and changes in coagulation-anticoagulant indexes were measured 6 h after injection of LPS or normal saline. Changes in malondialdehyde (MDA) content, wet and dry weight (W/D) ratio and morphometry of pulmonary tissue as well as PMN sequestration in the lung were also measured.

RESULTS(1) When compared with the control group, expression of PMNCD18 and MDA content were enhanced in the ALI group with a hypercoagulable state (all P<0.01) and an increased W/D ratio (P<0.05). Histopathological morphometry in the lung tissue showed higher PMN sequestration, wider alveolar septa; and lower alveolar volume density (V(V)) and alveolar surface density (S(V)), showing significant difference (P<0.01). (2) When compared with the ALI group, the expression of PMN-CD18, MDA content, and W/D ratio were all lower in Tanshinone II A treatment group (P<0.05) with ameliorated coagulation abnormality (P<0.01). Histopathological morphometry in the lung tissue showed a decrease in the PMN sequestration and the width of alveolar septa (both P<0.01), and an increase in the V(V) and S(V) (P<0.05, P<0.01).

CONCLUSIONTan II A plays a protective role in LPS-induced lung injury in rats through improving hypercoagulating state, decreasing PMN-CD18 expression and alleviating migration, reducing lipid peroxidation and alleviating pathological changes.

Animals ; Blood Coagulation ; drug effects ; CD18 Antigens ; analysis ; Diterpenes, Abietane ; Drugs, Chinese Herbal ; pharmacology ; Female ; Lipopolysaccharides ; toxicity ; Lung ; drug effects ; pathology ; Male ; Malondialdehyde ; analysis ; Phenanthrenes ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the toxicological mechanism of hydroquinone in human bronchial epithelial cells and to investigate whether DNA polymerase beta is involved in protecting cells from damage caused by hydroquinone.

METHODSDNA polymerase beta knock-down cell line was established via RNA interference as an experimental group. Normal human bronchial epithelial cells and cells transfected with the empty vector of pEGFP-C1 were used as controls. Cells were treated with different concentrations of hydroquinone (ranged from 10 micromol/L to 120 micromol/L) for 4 hours. MTT assay and Comet assay [single-cell gel electrophoresis (SCGE)] were performed respectively to detect the toxicity of hydroquinone.

RESULTSMTT assay showed that DNA polymerase beta knock-down cells treated with different concentrations of hydroquinone had a lower absorbance value at 490 nm than the control cells in a dose-dependant manner. Comet assay revealed that different concentrations of hydroquinone caused more severe DNA damage in DNA polymerase beta knock-down cell line than in control cells and there was no significant difference in the two control groups.

CONCLUSIONSHydroquinone has significant toxicity to human bronchial epithelial cells and causes DNA damage. DNA polymerase beta knock-down cell line appears more sensitive to hydroquinone than the control cells. The results suggest that DNA polymerase beta is involved in protecting cells from damage caused by hydroquinone.

Bronchi ; cytology ; drug effects ; Cells, Cultured ; Comet Assay ; Cytotoxins ; toxicity ; DNA Damage ; DNA Polymerase beta ; antagonists & inhibitors ; physiology ; Epithelial Cells ; cytology ; drug effects ; Humans ; Hydroquinones ; toxicity ; RNA Interference