摘要
目的探讨脂肪干细胞(adipose tissue derived stem cells,ADSCs)联合巴戟天提取物(Morinda officinalis How,MOH)对B6.MRL-Faslpr/Nju(B6.MRL/lpr)狼疮小鼠肾损伤及免疫炎性因子的影响。方法取C57BL/6J(C57)小鼠腹股沟脂肪分离培养ADSCs,CCK-8检测MOH对ADSCs细胞活性的影响。将10只C57小鼠设为正常组,40只B6.MRL/lpr小鼠随机分为模型组、ADSCs组、MOH组、ADSCs+MOH组。正常组和模型组灌胃生理盐水;ADSCs组尾静脉注射ADSCs(1×105个);MOH组灌胃MOH(10 mg·kg-1);ADSCs+MOH组应用ADSCs和MOH联合治疗,共8周。ELISA法检测血清中抗dsDNA抗体浓度变化,检测肾组织匀浆液中的肿瘤坏死因子-α(TNF-α)、白介素(IL)-17、IL-10、IL-6的表达水平,考马斯亮蓝法检测24 h尿蛋白的浓度变化;HE染色观察肾脏病理组织学变化。结果MOH浓度在20~200μg·mL-1范围内可以促进ADSCs增殖(P<0.01)。与正常组比,模型组抗dsDNA抗体、尿蛋白、TNF-α、IL-17、IL-10、IL-6明显升高(P<0.01);与模型组比,ADSCs组抗dsDNA抗体、尿蛋白、TNF-α、IL-17、IL-6均明显降低(P<0.01)。MOH组抗dsDNA抗体、尿蛋白、IL-17、IL-10、IL-6均显著降低(P<0.01)。ADSCs+MOH组抗dsDNA抗体、尿蛋白、TNF-α、IL-17、IL-10、IL-6均显著降低(P<0.01)。肾脏病理检查显示ADSCs、MOH、ADSCs+MOH均可明显改善B6.MRL/lpr小鼠的肾脏病理结构,减轻肾小球细胞增生,而ADSCs+MOH联合治疗效果较好。结论ADSCs、MOH、ADSCs+MOH均可改善B6.MRL/lpr小鼠肾脏病理结构,调节肾脏的免疫炎性因子,降低抗dsDNA抗体、尿蛋白的浓度,而ADSCs+MOH联合治疗效果最明显。MOH在体外可以促进ADSCs细胞的活性。ADSCs、MOH联用治疗B6.MRL/lpr小鼠具有协同作用。
Objective To investigate the effects of Morinda officinalis How(MOH)combined with ADSCs on renal injury and immunoinflammatory factors in B6.MRL/lpr lupus mice.Methods ADSCs were isolated from inguinal fat of C57 BL/6 J mice.The activity of ADSCs was measured by CCK-8 method.Ten C57 BL/6 J mice were set as normal group,and 40 B6.MRL/lpr mice were randomly divided into model group,ADSCs group,MOH group,ADSCs+MOH group.The normal group and the model group were fed with normal saline;ADSCs group mice were injected into the veins of the tails with ADSCs(1×105);rats in MOH group were given Morinda officinalis How(10 mg·kg-1);ADSCs+MOH group rats were given combination therapy.The treatments were last for 8 weeks.Concentration of antidsDNA antibody in mice’s serum was detected by ELISA,and the expression levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-17,IL-10 and IL-6 in renal homogenates were detected.Coomassie brilliant blue method were used to detect concentration of urine protein.HE staining was conducted to observe histopathological changes in kidney.Results The MOH concentration in the range of 20-200μg·mL-1 could promote the proliferation of ADSCs(P<0.01).Compared with the normal group,anti-dsDNA antibody,urinary protein,TNF-α,IL-17,IL-10 and IL-6 were significantly increased in the model group(P<0.01).Compared with the model group,antidsDNA antibody,urine protein,TNF-α,IL-17,IL-6 in ADSCs group and MOH group decreased(P<0.01);anti-dsDNA antibody,urine protein,TNF-α,IL-17,IL-10,IL-6 decreased in ADSCs+MOH group(P<0.01).HE showed that,ADSCs,MOH,ADSCs+MOH treatments can improve the renal pathological structure of B6.MRL/lpr mice,reduce glomerular cell proliferation,the effect of combination of ADSCs+MOH was the best.Conclusion ADSCs,MOH,ADSCs+MOH can improve the renal pathological structure of B6.MRL/lpr mice,regulate the immune inflammatory factors of the kidney,and decrease the concentrations of anti-dsDNA antibody and urine protein.The combination of ADSCs+MOH showed the most significant effects,and MOH can promote the activity of ADSCs in vitro.Therefore,ADSCs and MOH may simultaneously cure the B6.MRL/lpr mice by synergistic effect.
作者
陈美玲
邵艳华
唐佩华
苏俊芳
赵婷秀
郑嘉隆
张发扶
邝枣园
CHEN Meiling;SHAO Yanhua;TANG Peihua;SU Junfang;ZHENG Jialong;ZHANG Fafu;KUANG Zaoyuan(School of Basic Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2019年第9期1098-1104,共7页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
广州中医药大学高水平大学建设项目(A1-AFD018171Z11009)