Endoplasmic Reticulum Stress In Idiopathic Pulmonary Fibrosis

PART1:Effects of ERS on Bleomycin-Induced Pulmonary Fibrosis in C57BL/6MiceObjectiveTo determine the effects of tunicamycin induced ERS and Salubrinal blocked ERS pathway on bleomycin-induced pulmonary fibrosis in C57BL/6mice, and discuss the mechanisms.Materials and Methods1. We used a single dosage intratracheal injection in mice.2. The fibrosis degree was determined pathologically by using the Ashcroft scoring method and biochemically by hydroxyproline assay in lung tissue.3. To collect bronchoalveolar lavage fluid, the differential inflammatory cells were counted in BALF and the concentrations of TGF-βand MMP2in BALF were determined by ELISA.4. The expression of CHOP、GRP78、α-SMA and E-cadherin was detected by Immunohistochemistry and Western blot.5. The expression of XBP1、Caspase3、α-SMA and TGF-β mRNA in lung tissue was detected by Real-time PCR.Results1. Compare to the mice administrated by bleomycin and the control group, the Aschcroft score and hydroxyproline content were significantly increased in the mice administered tunicamycin, while salubrinal decreased the effects.2. Compare to the mice administrated by bleomycin, treatment with tunicamycin significantly increased the level of TGF-βand MMP2in BALF, while salubrinal decreased the effects.3. Compared to the bleomycin alone group, treatment with tunicamycin significantly increased the level of CHOP、GRP78、α-SMA and E-cadherin in the lung tissue, while salubrinal decreased the effects.4. Treatment with tunicamycin significantly increased the level of XBP1、Caspase3、α-SMA and TGF-β1in the lungs of mice.ConclusionAdministration of tunicamycin can aggravate bleomycin-induced pulmonary fibrosis in mice, while salubrinal decreased the effects. The mechanisms were possibly by inducing alveolar epithelial cell (AEC) apoptosis and inducing epithelial-mesenchymal transition. PART2:Effects of ERS on IPF PatientsObjectiveTo observe the expression of ERS associated proteins on IPF patients’lung tissue and plasma, discuss the mechanism and to investigate the application of biomarkers like CK18and cCK18in clinical diagnosis and treatment.Materials and Methods1. The apoptosis of AECs was determined pathologically by using TUNEL method;2. The expression of α-SMA、GRP78、CHOP and E-cadherin was detected by immunohistochemical method.3. The expression of CK18(M65) and cCK18(M30) in IPF patients’plasma was detected by ELISA method.Results1. Enhanced green fluorescence proteins was observed at the AECs region of the IPF patients’ lung.2. α-SMA、GRP78、CHOP were obviously expressed in IPF patients’lung tissue, while the expression of E-cadherin was obscure;3. The ratio of M30and M65was significantly decreased in IPF patients’plasma after12months of treatment with pirfinitone, and the expression of M30was lower in IPF patients than in other DPLD patients.ConclusionThe detection of CK18(M65) and cCK18(M30) have important clinical value in diagnosis and differential diagnosis in IPF patients. The AECs region of the IPF patients’ lung had marked apoptosis. PART3:The Safety of Lianhuaqingwen Preparations in Clinical Use:A Systematic ReviewObjectiveTo evaluate the safety of lianhuaqingwen preparations in clinical use.Materials and MethodsWe performed a systematic search of CBM, CNKI, Cochrane Central, MEDLINE and EMbase for trials published between January2003and May2013.ResultsFourty randomized controlled clinical trials including2592cases of the experimental group and2314cases of the control group were evaluated. The experimental group had63cases with adverse reactions(2.4%).While in the control group,100cases had adverse reactions(4.3%). The relative risk of the incidence of adverse reactions in the lianhuaqingwen preparations group was lower than the control group [RR=0.562,95%CI (0.412-0.767)].In the subgroup analysis of24trails, the total adverse effects RR was0.62,95%CI (0.46,0.82),and the adverse reactions of digestive system RR was0.70,95%,CI(0.50,97).The results suggested that the incidence of digestive system adverse reactions was lower in Lianhuaqingwen preparations group.ConclusionThe common adverse reactions of Lianhuaqingwen preparations were gastrointestinal and dermatological reactions.The incidence of those adverse reactions in Lianhuaqingwen preparations group was significantly lower than the control group.