ObjectiveTo observe the influence of heat shock protein 27 (HSP27) sensibilization to retinal ganglion cells (RGC) apoptosis of rats. MethodsThirty-five female Wistar rats were randomly divided into HSP27 sensibilization group (15 rats), borate buffer solution (BBS) control group (15 rats) and normal group (5 rats). The rats in HSP27 sensibilization group were received hypodermic injection in rear limb with 100 μg HSP27 and complete freund adjuvant, intraperitoneal injection with 1 μg pertussis toxin. The BBS control group received the same volume of BBS at the same site. The normal group received no intervention. The intraocular pressure was measured 3 days before injection and 1, 2, 4, 6, 8 weeks after injection. Four, 6 and 8 weeks after injection, the retinal frozen sections was made to observe RGC apoptosis by terminal-deoxynucleoitidyl transferase mediated nick end labeling. The anti-HSP27 level in serum and cerebrospinal fluid were detected by enzyme linked immunosorbent assay. ResultsThere was no obvious change of intraocular pressure in rats in 3 groups before injection (P>0.05). RGC apoptosis was observed in HSP27 sensibilization group 4 weeks after injection, and increased significantly at 6 weeks after injection. There was no RGC apoptosis in BBS control group and normal group. The level of anti-HSP27 in serum and cerebrospinal fluid of HSP27 sensibilization group occurred at 4 and 6 weeks after injection respectively, decreased with prolongation of injection time. Compared with BBS control group and normal group, the RGC apoptosis rate, anti-HSP27 level in serum and cerebrospinal fluid of HSP27 sensibilization group were significantly increased (P<0.05). There was no significant difference of the RGC apoptosis rate, anti-HSP27 level in serum and cerebrospinal fluid between BBS control group and normal group (P>0.05). ConclusionsHSP27 sensibilization could promote the RGC apoptosis. The variation trend of anti-HSP27 level in cerebrospinal fluid is consistent with the RGC apoptosis.
ObjectiveTo investagte the effects of single immunoglobin IL-1 receptor related protein (SIGIRR) on inflammation induced by cigarette smoke extract (CSE) in A549 cells derived from mouse alveolar epithelial cells. MethodsA549 cells were divided into a control group and an over-expressed SIGIRR group. Eukaryotic expression vectors pcDNA3.1(+) constructed with SIGIRR cDNA were transiently transfected into A549 cells, in which SIGIRR was forced to be over-expressed. The expression level of SIGIRR after transfection was detected with Western blot and RT-PCR method. After stimulated by CSE in both groups, the protein level of IL-6 was detected by ELISA, the transcriptional activity of cyclooxygenase-2(COX-2) was detected by dual-luciferase reporter assay system, and the release of reactive oxygen species (ROS) was measured by chemiluminescence method. ResultsThe expression level of SIGIRR increased significantly in A549 cells transfected with SIGIRR vectors. The COX-2 expression and the levels of ROS and IL-6 were significantly increased in the control group after CSE stimulation. Nevertheless, in the over-expressed SIGIRR group, the COX-2 expression and the release of ROS was reduced while the protein level of IL-6 was down-regulated compared with the control group(P < 0.05). ConclusionsUp-regulated SIGIRR expression can suppress the levels of ROS, COX-2 and IL-6 in A549 cells stimulated by CSE. It suggests that SIGIRR can inhibit airway inflammation caused by smoking.