热休克蛋白27致敏对大鼠视网膜神经节细胞凋亡的影响

The influence of heat shock protein 27 sensibilization to retinal ganglion cells apoptosis of rats

目的 观察热休克蛋白27（HSP27）致敏对大鼠视网膜神经节细胞（RGC）凋亡的影响.方法 35只雌性Wistar大鼠分为HSP27致敏组、硼酸盐缓冲液（BBS）对照组、正常组,分别为15、15、5只大鼠.给予HSP27致敏组大鼠后肢皮下注射100μg的HSP27和完全弗式佐剂,腹腔注射1μg百日咳毒素.BBS对照组大鼠于相同部位注射等量BBS.正常组大鼠不作任何处理.注射前3d及注射后3d,1、2、4、6、8周,检测大鼠双眼眼压.注射后4、6、8周行视网膜冰冻切片,采用原位末端标记法检测RGC的凋亡情况,计算细胞凋亡率;采用酶联免疫吸附试验检测大鼠血清和脑脊液中HSP27抗体的含量.结果 HSP27致敏组、BBS对照组、正常组大鼠注射前后眼压均无明显波动,差异均无统计学意义（P＞0.05）.HSP27致敏组于注射后4周即可见RGC凋亡,注射后6周凋亡的RGC明显增多.BBS对照组及正常组均未见RGC凋亡.HSP27致敏组大鼠血清、脑脊液中HSP27抗体含量分别于注射后4、6周出现高峰,然后随着注射时间延长而递减.注射后4、6、8周,HSP27致敏组RGC凋亡率、血清中HSP27抗体含量、脑脊液中HSP27抗体含量分别与BBS对照组、正常组比较,差异均有统计学意义（P＜0.05）;BBS对照组RGC凋亡率、血清中HSP27抗体含量、脑脊液中HSP27抗体含量与正常组比较,差异无统计学意义（P＞0.05）.结论 HSP27致敏可促进大鼠RGC的凋亡.脑脊液中HSP27抗体含量与RGC凋亡趋势一致.

Objective To observe the influence of heat shock protein 27 （HSP27） sensibilization to retinal ganglion cells （RGC） apoptosis of rats.Methods Thirty-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.Results There 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）.Conclusions HSP27 sensibilization could promote the RGC apoptosis.The variation trend of antiHSP27 level in cerebrospinal fluid is consistent with the RGC apoptosis.