1.Cerebral and lung c-fos protein expression and neuroimmunomodulation in asthmatic rats
Weiwen FAN ; Zhijun YANG ; Rongcheng LUO ; Ling WEI ; Huojin DENG
Chinese Journal of Tissue Engineering Research 2005;9(32):234-236
BACKGROUND: Increasing evidence identifies the immune system not as an isolated system with automodulations, but one that interacts with the central nervous system.OBJECTIVE: To investigate the distribution of c-fos expression in the lung and brain tissues of asthmatic rats and explore is significance.DESIGN: Randomized controlled study.SETTING: Department of Oncology, Southern Hospital, and Department of Neurosurgery, Zhujiang Hospital, Southern Medical University.MATERIALS: The experiment was conducted Department of Oncology,Southern Hospital, and Department of Neurosurgery, Zhujiang Hospital,Southern Medical University, between January and August 2004. Fourteen healthy male rats were randomized into experimental group (n=10) and control group (n=4).METHODS: On the first day of experiment, the rats in experimental group received intraperitoneal injection of 2 mL of the suspension containing 10 mg albumen, 200 mg aluminum hydroxide powder and inactivated pertussis vaccine (5×109), and subjected to inhalation of ultrasonically atomized 10 g/L albumen from on the 15th day, 2 times per hour for totally 3 days, to induce asthma in the rats. The rats in the control group received intraperitoneal injection of 2 mL normal saline on the 1st day and inhalation of ultrasonically normal saline on the 15th day, 30 mL a day for totally 3 days. The lung and brain tissues of all the anesthetized rats were fixed by perfusion, and immunohistochemical method with ovin-biotin-peroxidase complex and imaging analysis system were used to observe the distribution of Fos protein in the lung and brain.MAIN OUTCOME MEASURES: Distribution of c-Fos protein in lung and cerebrum.c-Fos in the lung and brain tissues was obviously higher in asthmatic group than in the control group (P < 0.05), located mainly in the parietal-fontal cortex, limbic forebrain (cingulum cortex, pyriform cortex and central amygdaloid nucleus and so on), thalamus paraventricular nucleus, hypothalamus paraventricular nucleus, supraoptic nucleus, lateral region of the hypothalamus, hypothalamus periventricular nucleus, nucleus of solitary tract,area postrema and ventrolateral medulla. No obvious Fos expression was observed in the cerebellum. A large number of c-Fos-positive cells were observed in the wall of the airway and lungs in asthmatic rats, mainly distributing in the mucous membrane and submucous layer and around the smooth muscles; in the control rats, no positive cells or only occasional cells with weak c-Fos positivity were found in the wall of the airway and lungs.CONCLUSION: c-Fos expression increases obviously not only in the lungs of asthmatic rats, but also in medullar and its ascending projecting nuclei (hypothalamus, amygdala and so on), suggesting that the expression of protooncogene c-fos might be closely related with neuroimmunomodulation in asthma.
2.Rapid detection of methicillin-resistant staphylococci by DNA probe
Huizhen FAN ; Huapeng YU ; Wenjie HUANG ; Huojin DENG
Chinese Journal of Clinical Laboratory Science 2006;0(05):-
Objective To establish a dot blot hybridization technique for rapid detection of staphylococci and methicillin-resistant staphylococci.Methods Three pairs of primers were designed according to nuc gene of staphylococcus aureus,mecA gene of methicillin-resistance,tuf gene of staphylococci.Specific DNA probes were synthesized by polymerase chain reaction and labeled with biotin.The bacterial DNA inoculated on nitrocellulose filter was hybridized with these probes.The sensitivity and specificity were detected.Results The DNA probes with 270bp,310bp and 370bp were amplified by the three pairs of primers respectively.The probes were specific.Among 50 clinical isolates of staphylococcus aureus tuf and nuc gene were all positive and mecA gene in 22 isolates were positive.Positive rate of tuf,nuc and mecA gene in 30 staphylococcus epidermidis were 100%,0 and 30% (9/30) respectively.No hybridization in other non-staphylococci occurred.The established method could detect as low as 1ng of bacterial DNA.Conclusion The dot blot hybridization is of high value in rapid,effective identification of methicillin-resistant staphylococci.
3.Curcumin suppresses cigarette smoke extract-induced oxidative stress through PPARγ/ NF-κB pathway in human bronchial epithelial cells .
Tao ZHU ; Chanmei SHI ; He LI ; Jing HE ; Yanli YANG ; Qin WANG ; Xinyu DENG ; Yanqiao WU ; Jing WANG ; Yan ZHAO ; Huojin DENG
Journal of Southern Medical University 2018;38(10):1209-1214
OBJECTIVETo investigate the effect of curcumin against cigarette smoke extract (CSE)- induced oxidative stress in human bronchial epithelial cells and explore the underlying mechanism.
METHODSHuman bronchial epithelial cell line 16HBE was treated for 24 h with curcumin, CSE, CSE + curcumin, and CSE + curcumin with transfection by a short hairpin RNA targeting PPARγ (shPPARγ). MTT assay was used to observe the changes in the cell viability after the treatments. Quantitative real-time PCR was performed to detect the mRNA expressions of tumor necrosis factor- (TNF-), iNOS and PPARγ in the cells, and the protein expressions of iNOS, PPARγ and the phosphorylation of NF-κB p65 were detected using Western blotting.
RESULTSThe treatments did not cause significant changes in the cell viability. Exposure to CSE for 24 h significantly lowered PPARγ expression and increased TNF- and iNOS expressions and phosphorylation of NF-κB p65 in the cells. The effects of CSE were significantly suppressed by curcumin, but transfection of the cells with shRNA-PPARγ obviously abrogated the suppressive effects of curcumin.
CONCLUSIONSCurcumin suppresses CSE-induced oxidative stress and inflammation via the PPARγ/NF-κB signaling pathway in 16HBE cells, suggesting the potential of curcumin in the treatment of chronic obstructive pulmonary disease.