炎症牙龈中环孢素A对基质金属蛋白酶表达的影响

Effects and significance of cyclosporine A on cyclophilin A, MMPs, p-ERK1/2, and p-IκB expressions in gingival inflammatory condition

目的:探讨牙龈炎症条件下环孢素A(cyclosporine A,CsA)对亲环素A(cyclophilin A,CyPA)、MMPs及p-ERK1/2、p-IκB表达的影响及意义。方法:24只8w雄性C57BL/6j小鼠适应性饲养lw后随机分成4组:对照组,结扎组,CsA组,结扎+CsA组,每组6只小鼠。结扎+CsA组:用5. 0无菌医用缝合丝线结扎左、右上颌第一磨牙牙颈部,7d后,腹腔内每日注射1次CsA (50mg/kg/d),共21d;结扎组:结扎方法同结扎+CsA组,7d后腹腔内每日注射1次等量0. 9%NaCL溶液,共21d;CsA组:实验开始第8d,腹腔内每日注射1次CsA (50mg/kg/d),共21d;对照组:不作任何处理,饲养条件同前3组。用HE染色检测牙龈组织增生情况,免疫组化实验检测牙龈组织中的CyPA的表达,用Western Blot检测牙龈组织中的Pro-MMP-2,-9、MMP-2,-9表达以及p-ERK1/2与ERK1/2、p-IκB与IκB的比值。结果:H E染色结果显示对照组中的组织结构完整;结扎组中牙齿的邻接区和根尖区炎症浸润,结扎+CsA组的骨损伤少于结扎组,结扎+CsA组相较于其他3组牙龈增生最明显。免疫组化结果显示结扎组CyPA表达最高,CsA组低于对照组,而结扎+CsA组则略高于对照和组和CsA组。Western Blot的结果显示结扎组Pro-MMP-9、MMP-9、Pro-MMP-2、MMP-2表达最高,其次为结扎+CsA组;结扎组的p-ERK1/2/ERK1/2及p-IκB/IkB比值最高,其他3组中,结扎+CsA组的p-ERK1/2/ERK1/2及p-IkB/IkB比值均高于对照组和CsA组。结论:在牙龈炎症条件下,CsA及炎症因子促进细胞外基质(Extracellular Matrix,ECM)合成增加;CsA抑制了MMPs的表达,导致ECM降解减少,并且CsA是通过阻断CyPA与CD147的相互作用,影响ERK及IkB的磷酸化,导致MMPs的表达减少和活性降低。

Objective:To investigate effects and significance of cyclosporine A(CsA)on cyclophilin(CyPA),MMPs,p-ERK1/2,and p-IκB expressions in gingival inflammatory condition.Methods:Twenty-four 8-week male C57 BL/6 j mice were randomly divided into 4 groups after 1 week of adaptive feeding:control group,ligation group,Cs A group,ligation+Cs A group,6 mice in each group.The control group had normal feeding and did not receive any treatment.In both ligation group and ligation+CsA group,5.0 sterile medical sutures were used to ligation the left and right maxillary first molars,seven days later,animals in ligation+Cs A group were intraperitoneally injected with Cs A(50 mg/kg/d),and animals in ligation group were intraperitoneally injected with the same amount of normal saline.Animals in Cs A group,only Cs A(50 mg/kg/d)was used to intraperitoneally inject.All 21 days later,the maxillary specimens were dissected and fixed in neutral formalin fixative.HE staining was used to detect the proliferation of gingival tissues.Immunohistochemistry was used to detect the expression of Cy PA in gingival tissues.Western Blot was used to detect Pro-MMP-2,-9,MMP-2,-9 expressions and the ratios of p-ERK1/2/ERK1/2,p-IκB/IκB in gingival tissues.Results:HE staining showed that the tissue structure in the control group was intact.the adjacent area and apical area of the teeth in the ligation group were inflammatory infiltration,and there was bone absorption.the Cs A group had slight gingival overgrowth.the Cs A+ligation group had obvious gingival hyperplasia and a small amount of bone absorption.The results of immunohistochemistry showed that the expressive level of Cy PA was highest in the ligation group compared with other groups,and the expression of Cy PA in the ligation+Cs A group was lower than that in the ligation group.The results of Western Blot showed that the expressive levels of Pro-MMP-2,-9,MMP-2,-9 were highest in the ligation group,and the expressive level in the ligation+CsA group was significantly inhibited compared with the ligation group.The ratios of p-ERK1/2/ERK1/2 and p-IκB/IkB in the ligation group were highest compared with other groups,and the ratios of p-ERK1/2/ERK1/2 and p-IκB/Ik B in the ligation group+CsA group were higher than that in control group and Cs A group.Conclusions:In gingival inflammtory condition,Cs A and inflammatory cytokines can augment extracellular matrix,on the other hand,Cs A may inhibits the expression and activity MMPs through inhibition of Cy PA-CD147 interaction and reduces the phosphorylation of ERK1/2 and IκB.