碘乙酸钠诱导的大鼠膝骨关节炎模型肠道菌群变化及其潜在影响

目的:研究碘乙酸钠诱导的大鼠膝骨关节炎(KOA)模型肠道菌群变化情况,探究肠道菌群与KOA的潜在联系。方法:将SPF级大鼠分为空白对照组和KOA组,通过膝关节局部注射碘乙酸钠制造大鼠KOA模型。通过16SrDNA检测大鼠粪便中的肠道菌群变化情况,蛋白印记法检测结肠组织中NF-κB和TNF-α蛋白表达,RT-qPCR检测结肠组织中Caspase-1和IL-1β基因表达情况,苏木精-伊红染色观察结肠黏膜通透性变化,苏木精-伊红染色观察关节滑膜和软骨组织的病理变化情况。ELISA检测血液上清中LBP,IL-1β,TNF-α及IL-10炎症因子水平。结果:与空白对照组相比,从KOA组中筛选出明显变化的肠道菌群,以Akkermansia和疣微菌门变化最为显著,结肠黏膜通透性增加,结肠组织中NF-κB p65及TNF-α蛋白表达升高,结肠组织中Caspase-1和IL-1β基因表达升高,血液中促炎因子LBP,IL-1β及TNF-α蛋白含量升高,抗炎因子IL-10蛋白含量降低。结论:KOA病理状态下肠道菌群发生变化,结肠炎症反应增强,结肠炎症反应或许在提高全身炎症反应水平、促进与肠道菌群相关的KOA的病理进程中起到重要的桥梁作用。

Intestinal Flora Changes and Its Potential Effects in Rats with Knee Osteoarthritis Induced by Iodoacetate Sodium

Objective:To study the changes of intestinal flora in the model of knee osteoarthritis(KOA)induced by iodoacetate sodium and explore the potential relationship between intestinal flora and KOA.Methods:SPF rats were divided into blank control group and KOA group,KOA rat model was made by local injection of iodoacetic acid into the knee joint.Changes of intestinal flora in rat feces were detected by 16 SrDNA.Protein expression of NF-κB,TNF-αin colon tissue were detected by Western Blot.The gene expression of caspase-1,IL-1βin colon tissue were detected by RT-qPCR.The permeability of colonic mucosa was observed by HE staining.Pathological changes of synovial and cartilage tissue were observed by HE staining.Inflammatory factors of LBP,IL-1,TNF-α,IL-10 in serum of rats were detected by ELISA.Results:Compared with control group,changed intestinal flora were screened out from KOA group,akkermansia and verrucomicrobia were the most significant,permeability of colon mucosa was destructed,protein expression of NF-κB,TNF-αand gene expression of caspase-1,IL-1βin colon tissue were increased(P<0.05).Inflammatory factors of LBP,IL-1,TNF-αincreased in serum while IL-10 decreased(P<0.01).Conclusion:Under the pathological state of KOA,intestinal flora changes and colonic inflammatory response is enhanced.Colonic inflammatory response may play an important role in improving the level of systemic inflammatory response and promoting the pathological process of KOA related to intestinal flora.