白细胞介素22和p38 MAPK信号通路抑制骨关节结核骨质破坏的作用机制研究  

Study on the mechanism of IL-22 and p38 MAPK signaling pathways in inhibiting bone destruction in bone and joint tuberculosis

作　　者：盛杰 洪凯峰 米尔扎提·艾沙 唐伟 地里下提·阿不力孜 Sheng Jie;Hong Kaifeng;Mierzhati Aisha;Tang Wei;Dilixiati Abulizi(Department of Orthopedics,Xinjiang Uygur Autonomous Region Infectious Disease Hospital,Urumqi 830000,China;Department of Orthopedics,Urumqi Friendship Hospital,Xinjiang Uygur Autonomous Region,Urumqi 830049,China)

机构地区：[1]新疆维吾尔自治区传染病医院骨科,乌鲁木齐830000 [2]新疆维吾尔自治区乌鲁木齐市友谊医院骨科,乌鲁木齐830049

出　　处：《中国防痨杂志》2025年第4期454-459,共6页Chinese Journal of Antituberculosis

基　　金：新疆维吾尔自治区科技厅自然科学基金面上项目(2021D01A161);新疆维吾尔自治区“天山英才”医药卫生高层次人才培养计划(TSYC202301B087)。

摘　　要：目的:探讨白细胞介素22(IL-22)通过p38 MAPK信号通路影响骨关节结核骨质破坏的作用机制。方法:(1)选取2021年1—12月在新疆维吾尔自治区传染病医院骨科接受手术治疗的骨关节结核患者作为病例组,共58例;选取相同时间段该医院接受手术治疗的骨折患者作为对照组,共51例。收集研究对象的血清样本,使用酶联免疫吸附试验检测IL-22水平;收集骨组织样本,应用Western blot检测p38、p-p38、CatK和MMP9蛋白表达水平。(2)将小鼠单核巨噬细胞白血病细胞(RAW264.7)诱导为破骨细胞模型,实验分为6组,分别为破骨细胞组、破骨细胞+H37Rv组、破骨细胞+H37Rv+阴性对照组、破骨细胞+H37Rv+IL-22-shRNA组、破骨细胞+H37Rv+p38抑制剂组、破骨细胞+H37Rv+p38抑制剂+IL-22-shRNA组。通过TRAP染色法观察破骨细胞的活性,使用qRT-PCR和Western blot检测p38、CatK和MMP9的基因和蛋白表达水平。结果:(1)临床研究结果显示,对照组血清中IL-22的表达水平[中位数(四分位数)]为34.61(28.29,76.92)pg/ml,明显低于病例组[102.74(56.12,132.78)pg/ml],差异有统计学意义(Z=-3.840,P=0.001)。病例组p38、p-p38、CatK和MMP9蛋白表达水平分别为0.649±0.043、0.856±0.062、0.506±0.072,均明显高于对照组(分别为0.346±0.078、0.708±0.094、0.366±0.046),差异均有统计学意义(t值分别为-8.368、-3.203、-4.025,P值均<0.05)。(2)细胞实验结果显示,破骨细胞+H37Rv组每光镜视野(×100)TRAP染色阳性细胞个数为51.333±12.858,明显高于其他组,差异有统计学意义(F=26.270,P<0.001);破骨细胞+H37Rv组p-p38、CatK和MMP9蛋白表达水平明显升高,与之相比,在IL-22-shRNA和p38抑制剂干预后明显下调。结论:敲减IL-22抑制p38 MAPK信号通路可减轻骨关节结核的骨质破坏。Objective:o elucidate the mechanistic role of interleukin-22(IL-22)in mediating bone destruction in bone and joint tuberculosis via the p38 mitogen-activated protein kinase(MAPK)signaling pathway.Methods:(1)A total of 58 patients diagnosed with bone and joint tuberculosis who underwent surgical intervention at the Department of Orthopedics,Xinjiang Uygur Autonomous Region Infectious Disease Hospital between January and December 2021 were enrolled as the case group.A control group comprising 51 patients with fractures who underwent surgical treatment at the same institution during the same period was also established.Peripheral blood serum samples were collected from all participants,and IL-22 levels were quantified using enzyme-linked immunosorbent assay(ELISA).Additionally,bone tissue specimens were obtained for protein expression analysis.The expression levels of p38 MAPK,phosphorylated p38(p-p38),cathepsin K(CatK),and matrix metalloproteinase-9(MMP9)were determined via Western blotting.(2)Mouse monocyte-macrophage leukemia cells(RAW264.7)were differentiated into osteoclast-like cells and subsequently divided into six experimental groups:osteoclast group,osteoclast+H37Rv group,osteoclast+H37Rv+negative control group,osteoclast+H37Rv+IL-22-shRNA group,osteoclast+H37Rv+p38 inhibitor group,and osteoclast+H37Rv+p38 inhibitor+IL-22-shRNA group.The osteoclastogenic activity was assessed using tartrate-resistant acid phosphatase(TRAP)staining,while gene and protein expression levels of p38 MAPK,CatK,and MMP9 were quantified using quantitative real-time polymerase chain reaction(qRT-PCR)and Western blotting.Results:(1)Clinical findings:The serum IL-22 levels in the control group(median(quartiles):34.61(28.29,76.92)pg/ml)were significantly lower than those in the case group(102.74(56.12,132.78)pg/ml),with a statistically significant difference(Z=-3.840,P=0.001).The protein expression levels of p38,p-p38,CatK,and MMP9 in the case group were 0.649±0.043,0.856±0.062,and 0.506±0.072,respectively,all of which were signifi

关 键 词：分枝杆菌 结核 结核 骨关节 破骨细胞 白细胞介素类 

分 类 号：R529.2[医药卫生—内科学]