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The Effect of Heme Oxygenase-1 on the Apoptosis and the Level of CHOP in NHBE Cells of COPD Induced by CSE

Author: ZhangLi
Tutor: LuoBaiLing
School: Central South University
Course: Department of Respiratory Medicine
Keywords: NHBE COPD CSE HO-1 CHOP
CLC: R363
Type: Master's thesis
Year: 2011
Downloads: 104
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Abstract

Objective:The most important pathological feature of chronic obstructive pulmonary diseases(COPD) is chronic bronchitis and pulmonary emphysema.The prominent pathological changes are the gradually lost of barrier function of bronchial epithelial and the degeneration、death、fall off of the epitheial cells. Endoplasmic reticulum stress(ERS)-mediated apoptosis pathway is a new apoptosis pathway. CHOP/GADD153(CCAAT/enhancer binding protein homologous protein) can be induced by ERS and its role is to block the cell division cycle,which may induce apoptosis. When in non-ER Stress, the expression of CHOP is rather low. When in ER stress,the expression of CHOP is greatly increased. When in the process of ERS, CHOP is a an important signaling molecule which convert antiapoptotic state to proapoptotic state. CHOP have an effect in ERS-induced apoptosis. Heme oxygenase-1(HO-1) is the most ubiquitous antioxidant enzyme, which can be induced by variety of noxious stimulation at high levels. There are many effects for HO-1, such as anti-inflammatory, antioxidant, anti-apoptosis, anti-proliferative effects. In many diseases such as bronchial asthma, pulmonary fibrosis,heart and lung ischemia-reperfusion injury,HO-1 plays a cell protective effection. Smoking is an important dangerous factor of COPD. Cigarette Smoke induce Normal human bronchial epithelial cells (NHBE cells) apoptosis through ERS. Cigeratte Smoke, which have many toxic ingredients and oxygen free radicals, can induce NHBE cells apoptosis through ESR. This research focused on cigeratte smoke extract (CSE) intervention of NHBE cells apoptosis by ESR, Our reaserch is focused on the level of CHOP,which is induced by the CSE. In the reaserch,we test the antiapoptosis effect of HO-1,and see if we can downregulate the expression of CHOP through the inducement of expression of HO-1. Whether the HO-1 can increase cells ability of antiapoptosis and resistent to ERS. We try to find more experimental evidences for the clinic treatment of COPD.Methods:The experiment was divided into four groups, including CSE group, CSE+hemin group, CSE+Znpp group, Control group. NHBE cells were cultured in vitro. It was treated with certain kind of concentration of CSE and Hemin and Znpp. The expression levels of CHOP and HO-1 were detected by Western blot. Flow cytometric cytology method and Hoechst 33258 were used to analyze NHBE cells apoptosis.Results:1.In the first two groups, including CSE group、CSE+Hemin group, HO-1 protein are significant expression. HO-1 expression in CSE group and CSE+Hemin group are higher than control group and CSE+Znpp group (p<0.05).2.In the control group、CSE+Hemin group、CSE group、CSE+Znpp group, the expression of CHOP is from low to high.3.Typical characteristics of apoptosis were found in the CSE group、CSE+Hemin and CSE+Znpp group. All groups were stained with Hoechst33258. Cells in CSE group, CSE+Hemin group and CSE+Znpp group have showed strong nuclear staining, particle block of fluorescence and obvious morphology cracking. Flow cytometry showed that the first three groups(including CSE group、CSE+Hemin group and CSE+Znpp group), the detection of apoptosis rates are higher than the control group(P<0.05). CHOP protein expression of apoptosis are significantly increased, which are compared with the control groups. The apoptosis rate in the four groups which added in CSE rank from CSE+Znpp group. CSE group, CSE+hemin group, to Control group shows:20.85±1.63%、12.73±1.48%、6.09±1.39%、0.88±0.42%. Apoptosis rates were decreasing (P<0.05), which were consistent with the expression of CHOP.Conclusions:1.CSE can induce NHBE cells apoptosis, with time and concentration dependent. 2.CHOP participates in the CSE-induced ERS,which make the NHBE cells apoptosis.3. The expression of HO-1 can restrain the NHBE cells apoptosis induced by cigarette smoke,this effect might be achieved by inhibiting the expression of CHOP.

The Effect of Heme Oxygenase-1 on the Apoptosis and the Level of CHOP in NHBE Cells of COPD Induced by CSE

Abstract

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