青蒿琥酯联合胰岛素对1型糖尿病合并牙周炎大鼠股骨改建的影响及机制

Effect of artesunate combined with insulin on femur remodeling in rats with type 1 diabetes mellitus and periodontitis

目的观察青蒿琥酯(ART)联合胰岛素对1型糖尿病合并牙周炎大鼠股骨改建的影响,并探讨其作用机制。方法取成年雄性SD大鼠40只,标准饲料喂养。将大鼠随机分为模型组32只与正常对照组8只。模型组采用腹腔注射链脲佐菌素的方法制作1型糖尿病模型,正常对照组一次性腹腔注射等量柠檬酸—柠檬酸钠注射液。模型组造模成功后,采用双侧上颌第一磨牙牙颈部结扎合并龈沟内接种牙龈卟啉单胞菌液制作牙周炎模型。将模型组随机分为模型对照组、ART干预组、INS干预组、ART+INS干预组,ART干预组给予ART注射液50 mg/kg灌胃,INS干预组给予重组人胰岛素注射液6 U/kg腹腔注射,ART+INS干预组给予50 mg/kg ART注射液灌胃及6 U/kg重组人胰岛素注射液腹腔注射,正常对照组及模型对照组使用等量生理盐水干预,连续4周。大鼠麻醉处死,逐层剥离双侧股骨,左侧股骨使用micro-CT测量平均骨密度(BMD);右侧股骨制成组织切片,采用HE染色法观察骨微结构改变,免疫组化染色法检测骨组织白细胞介素1β(IL-1β)、骨保护素(OPG)蛋白表达。结果与正常对照组相比,模型对照组BMD下降,骨小梁破坏严重,骨组织IL-1β表达升高、OPG表达下降(P均<0.05);与模型对照组相比,ART干预组、INS干预组、ART+INS干预组股骨BMD均升高,骨小梁增厚,骨组织IL-1β表达减少、OPG表达升高(P均<0.05);其中ART+INS干预组较ART干预组和INS干预组的股骨BMD升高,骨组织IL-1β降低、OPG增多(P均<0.05)。结论ART联合胰岛素可抑制1型糖尿病合并牙周炎大鼠股骨炎症反应及破骨改变,增强骨质结构,提升骨密度;其机制与介导IL-1β、OPG蛋白表达相关。

Objective To observe the effect of artesunate combined with insulin on femur remodeling in rats with type 1 diabetes mellitus and periodontitis and to investigate its mechanism.Methods Forty adult male SD rats were fed with standard diet.The rats were randomly divided into the model group(n=32)and normal control group(n=8).The rats in the model group were given intraperitoneal injection of streptozotocin(STZ)to make type 1 diabetes models,while the rats in the control group were given intraperitoneal injection of the same amount of citric acid-sodium citrate injection.In the model group,the periodontitis model was made by ligating the neck of bilateral maxillary first molars and inoculating P.gingivalis bacteria into gingival sulcus.The model group was randomly divided into the model control group,ART inter⁃vention group,INS intervention group,and ART+INS intervention group.ART intervention group was given art injection 50 mg/kg by gavage,INS intervention group was given recombinant human insulin injection 6 U/kg by intraperitoneal in⁃jection,ART+INS intervention group was given art injection 50 mg/kg by gavage and 6 U/kg by intraperitoneal injection.The normal control group and model control group were intervened with the same amount of normal saline for 4 weeks.The left femur was used to measure the mean bone mineral density(BMD);the right femur was made into tissue sections.The changes of bone microstructure were observed by HE staining,and the expression levels of IL-1βand osteoprotegerin(OPG)protein were detected by immunohistochemical staining.Results Compared with the normal control group,the BMD decreased,the trabecular bone was seriously damaged,the expression of IL-1βincreased,and the expression of OPG decreased in the model control group(all P<0.05).Compared with the normal control group,the BMD increased,the trabecular bone was thickened,the expression of IL-1βdecreased,and the expression of OPG increased in the ART in⁃tervention group,INS intervention group,and ART+INS intervention group(all P<0.05);the BMD of the ART+INS inter⁃vention group was higher,the expression of IL-1βwas lower,and the expression of OPG was higher as compared with those of the model control group(all P<0.05).Conclusion Artesunate combined with insulin can reduce the inflamma⁃tory reaction and osteoclastic changes,enhance the bone structure,and increase the average bone density of rats with type 1 diabetic mellitus and periodontitis by mediating the expression levels of IL-1βand OPG protein.