摘要
目的 探究MMP-3基因siRNA慢病毒干扰载体对大鼠骨性关节炎模型的影响。方法 制备SD大鼠OA模型,构建shRNA MMP-3 慢病毒表达载体,按照处理方法分为4组:模型对照组、假手术组、正常对照组和shRNA MMP-3载体组。每组为5只SD大鼠,模型对照组术后1月膝关节注射的是1 mL生理盐水,shRNA MMP-3组术后1月膝关节注射的是100 uL混合1 mL的生理盐水的shRNA MMP-3慢病毒载体。大体观察SD大鼠OA模型的膝关节,并进行番红染色和免疫组织化学染色。并进行相应的评分。结果(1)SD大鼠膝关节软骨的大体评分模型对照组、假手术组、正常对照组和shRNA MMP-3载体组的4组比较,SD大鼠膝关节OA软骨的破坏具有统计学差异(P < 0.05);(2)SD大鼠膝关节OA软骨的番红染色Makin评分的四组比较中具有统计学差异(P < 0.05),shRNA MMP-3载体组评分高于模型对照组,差异有统计学差异(q = 9.438,P < 0.01);(3)SD大鼠膝关节OA软骨的aggrecan蛋白的表达中,模型对照组、假手术组、正常对照组和shRNA MMP-3载体组的四组Aggrecan表达免疫组织化学染色评分具有统计学差异(P < 0.05),shRNA MMP-3载体组评分高于模型对照组,差异有统计学意义(q = 9.438,P < 0.01)。结论 shRNA MMP-3载体可以有效的保护Aggrecan的表达,减缓OA软骨的退变。
Objective To investigate the effect of siRNA lentivirus interference vector of MMP-3 gene on rat osteoarthritis model. Methods The modified SD rat OA model was prepared and the lentivirus vector of shRNA MMP-3 was constructed. According to the treatment methods,rats were divided into 4 groups:the model control group, the sham operation group, the normal control group and the shRNA MMP-3 lentiviral vector group. Five SD rats in each group were injected with 1 mL normal saline one month after operation in the model control group and 100 uL mixed with 1 mL normal saline shRNA MMP-3 lentiviral vector one month after operation in the shRNA MMP-3 group,The knee joint of SD rat OA model was observed and stained with Mandarin dyeing and immunohistochemistry,and the corresponding score was recorded. Results(1) SD rat knee cartilage gross score was compared among model control group and sham operation group,there was statistical difference in the destruction of OA cartilage of knee joint between normal control group and lentivirus carrier group with shRNA MMP-3(P < 0.05).(2) There was statistical difference among the four groups of the knee OA cartilage of SD rats(P < 0.05). The score of shRNA MMP-3 vector group was higher than that of model control group, and the difference was statistically significant(q = 9.438,P < 0.05).(3) In the expression of aggrecan protein in OA cartilage of knee joint of SD rats, the immunohistochemical staining scores of Aggrecan expression in the four groups of model control group, sham operation group, normal control group and shRNA MMP-3 carrier group were significantly different(P < 0.05). The score of shRNA MMP-3 vector group was higher than that of model control group, and the difference was statistically significant(q = 9.438,P < 0.05). Conclusion ShRNA MMP-3 vector can effectively protect the expression of Aggrecan and slow down the degeneration of OA cartilage.
作者
李勤学
李传静
王锋
LI Qin-xue;LI Chuan-jing;WANG Feng(Dept. of Laboratory Medicine,of Xiaogan Central Hospital, HubeiProvince, Xiaogan Hubei 432100,China;Dept. of Endocrinology,of Xiaogan Central Hospital, HubeiProvince, Xiaogan Hubei 432100,China)
出处
《昆明医科大学学报》
CAS
2019年第6期33-38,共6页
Journal of Kunming Medical University
基金
湖北省自然科学基金资助项目(2015CKC904)