摘要
目的观察miR-141及基质金属蛋白酶-16(MMP-16)对骨性关节炎(OA)疼痛及软骨损伤的影响,并探讨其部分调控机制。方法将30只健康雄性SD大鼠随机分为对照组10只和实验组20只,实验组大鼠利用十字韧带损伤法制作OA模型,分别于术前及手术2周和8周后采用冷痛阈法测定2组大鼠冷痛阈值,HE染色法观察2组大鼠软骨组织结构变化,实时定量PCR法检测大鼠背根神经节组织中miR-141表达情况,免疫组织化学法观察软骨组织中MMP-16蛋白表达情况。结果手术2,8周后,对照组大鼠对冷丙酮刺激的阳性反应次数未见明显变化(P均>0.05);实验组大鼠对丙酮刺激的阳性反应次数明显高于对照组(P<0.05),并且随着时间的延长,阳性反应次明显增加(P<0.05);HE染色显示,手术2周后实验组大鼠软骨细胞结构轻度紊乱伴少量炎性细胞浸润,同时细胞间质出现轻度纤维化;手术8周后,细胞形态基本消失,基质淡染,呈现广泛纤维化。手术2,8周后,实验组大鼠背根神经节组织中miR-141相对表达量和骨组织中MMP-16蛋白表达量均明显高于对照组(P均<0.05),且手术8周后明显高于手术2周后(P<0.05)。结论脊髓背角细胞中的miR-141在OA发展中调节疼痛的发生,而MMP-16促进OA的发展。
Objective It is to observe the effect of miR-141 and MMP-16 on pain and cartilage injury in osteoarthritis (OA), and to explore its regulation mechanism. Methods 30 healthy male SD rats were randomly divided into control group (n = 10) and experiment group (n = 20). The rats in the experimental group were used to establish OA models by anterior eruciate ligament transection. The cold hyperalgesia of all the rats in both groups were evaluated by cold hyperalgesia method before operation, in 2, 8 weeks after operation. The cartilage tissue structure was observed by HE staining. The expression of miR-141 in spinal cord dorsal horn was detected by real-time quantitative PCR (RT-PCR). The expression of MMP-16 was detected by immunohistochemistry (IHC). Results In 2 and 8 weeks after operation, no obvious change of the positive times of reaction to cold acetone simulation was found in the control group ( P 〉 0.05 ) , but the times in the experimental group were significantly more than that in the control group (P 〈0. 05) , with time increase, the times of positive reaction increased significantly (P 〈 0.05). HE staining showed that in 2 weeks after operation, the cartilage cell structure of the rats in the experiment group was disordered with inflammatory cell infiltration and a little fibrosis, in 8 weeks the cell structure disappeared with with pale cytoplasm and wide fibrosis. RT-PCR showed that the expression of miR-141 in the spinal dorsal horn cells has significantly increased. In 2 and 8 weeks after operation, the expression of miR- 141 in dorsal root ganglion tissue and MMP-16 protein in bone tissue in the experiment group were higher than that in the control group ( P 〈 0.05 ) , furthermore, the expression in 8 weeks after operation was higher than that in 2 weeks. Conclusion MiR-141 in the spinal dorsal horn cells regulates the pain in the development of OA, whereas MMP-16 promotes the development of OA.
出处
《现代中西医结合杂志》
CAS
2018年第4期347-350,411,共5页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
湖北省教育厅支持项目(B2016013)