舒筋健腰丸对腰椎黄韧带纤维化模型大鼠黄韧带中巨噬细胞M1/M2极化的影响  

Effect of Shujin Jianyao Pills (舒筋健腰丸) on M1/M2 Polarization of Macrophages in Ligamentum Flavum of Lumbar Ligamentum Flavum Fibrosis Model Rats

作　　者：冯蓬 高春雨[1] 李路广 李建国 高景华[1] 刘海英 FENG Peng;GAO Chunyu;LI Luguang;LI Jianguo;GAO Jinghua;LIU Haiying(Wangjing Hospital,China Academy of Chinese Medical Sciences,Beijing,100020;Guangzhou Baiyunshan Chenliji Phar-maceutical Factory Co.,Ltd)

机构地区：[1]中国中医科学院望京医院,北京100020 [2]广州白云山陈李济药厂有限公司

出　　处：《中医杂志》2023年第16期1684-1690,共7页Journal of Traditional Chinese Medicine

基　　金：中国中医科学院望京医院院级科研课题(2021-HX-7)。

摘　　要：目的 探讨舒筋健腰丸对腰椎黄韧带纤维化的影响及可能作用机制。方法 将30只SD大鼠随机分为正常组、模型组、阴性对照组、阳性对照组和舒筋健腰丸组,每组6只。除正常组外其余各组大鼠均采用腰椎失稳损伤法建立大鼠黄韧带纤维化模型。造模手术后72 h开始,舒筋健腰丸组给予舒筋健腰丸1.5 g/(kg·d)灌胃,其余各组给予生理盐水30 ml/(kg·d)灌胃,每日1次,持续8周。阴性对照组和阳性对照组灌胃的同时,分别于造模后即刻、2周、4周和6周于原切口皮下注射5 mg/ml PBS脂质体0.5 ml和5 mg/ml氯磷酸二钠脂质体0.5 ml。采用Masson染色观察大鼠黄韧带病理学情况,计算胶原容积分数(CVF),采用免疫组化染色法检测黄韧带中诱导型一氧化氮合酶(iNOS)、肿瘤坏死因子α (TNF-α)、小鼠含生长因子样模体黏液样激素样受体(F4/80)、转化生长因子β_(1)(TGF-β_(1))蛋白表达,采用免疫荧光双标法检测黄韧带M2巨噬细胞标记物F4/80和TGF-β_(1)表达。结果 与正常组相比,模型组、阴性对照组和舒筋健腰丸组Masson染色可见弹性纤维不同程度断裂,胶原纤维不同程度堆积,CVF显著升高,腰椎黄韧带iNOS、TNF-α、F4/80和TGF-β_(1)表达显著增多(P<0.01);与模型组相比,阳性对照组和舒筋健腰丸组胶原纤维堆积均减少,胶原纤维和弹性纤维交联松散,CVF显著降低,阳性对照组iNOS、TNF-α、F4/80和TGF-β_(1)表达显著减少,舒筋健腰丸组iNOS表达显著增多,TNF-α、F4/80和TGF-β_(1)表达显著减少(P<0.05或P<0.01);与阳性对照组相比,舒筋健腰丸组胶原纤维堆积相对较多,CVF显著升高,腰椎黄韧带iNOS、TNF-α、F4/80和TGF-β_(1)表达均显著增多(P<0.05或P<0.01)。免疫荧光结果显示,正常组和阳性对照组均未见F4/80与TGF-β_(1)表达,模型组F4/80与TGF-β_(1)大量表达并共定位,舒筋健腰丸组可见较多F4/80表达,但TGF-β_(1)表达较少。结论 舒筋健腰丸能够抑制大Objective To explore the effect of Shujin Jianyao Pills (舒筋健腰丸) on fibrosis of lumbar ligamentum flavum and the possible mechanism.Methods Thirty SD rats were randomly divided into normal group,model group,negative control group,positive control group and Shujin Jianyao Pills group,with six rats in each group.Lumbar instability injury method was used to establish rat models of ligamentum flavum fibrosis in all groups except the normal group.Seventy-two hours after the modeling,the Shujin Jianyao Pills group was given 1.5 g·kg^(-1)·d^(-1) of Shujin Jianyao Pills by intragastric administration,while the other groups were given 30 ml/kg^(-1)·d^(-1) of normal saline by intragastric administration,once a day for 8 weeks.Besides,in the negative control group and the positive control group,PBS liposomes(5 mg/ml,0.5 ml) and clophosphate liposomes(5 mg/ml,0.5 ml) were subcutaneously injected into the original incision respectively at once,2 weeks,4 weeks and 6 weeks after the modeling.The rat ligamentum flavum was evaluated histologically by Masson staining,and the collagen volume fraction(CVF) was calculated.Immunohistochemical staining was used to detect inducible nitric oxide synthase(iNOS),tumor necrosis factor-alpha(TNF-α),mouse growth factor-like phantom-containing mucus-like hormone-like receptor(F4/80) and transforming growth factor β_(1)(TGF-β_(1)) protein expression in ligamentum flavum.The expressions of F4/80 and TGF-β_(1),the markers of M2 macrophages in the ligamentum flavum,were detected by immunofluorescence double labeling.Results Compared to those in the normal group,the Masson staining of the model group,the negative control group and the Shujin Jianyao Pills group showed elastic fiber breakage and collagen accumulation,and the CVF,as well as the expression of iNOS,TNF-α,F4/80 and TGF-β_(1) in the lumbar ligamentum flavum increased significantly(P<0.01).Compared to those in the model group,the collagen fibers in the positive control group and Shujin Jianyao Pills were reduced,while th

关 键 词：黄韧带肥厚 黄韧带纤维化 腰椎 巨噬细胞极化 舒筋健腰丸 

分 类 号：R285.5[医药卫生—中药学]