羊肠线埋入对大鼠“足三里”穴区巨噬细胞CD68、肿瘤坏死因子-α及白细胞介素-1β的影响

Effect of catgut implantation on macrophage CD68,tumor necrosis factor-αand interleukin-1βin“Zusanli”(ST 36)region of rats

目的:观察大鼠“足三里”穴区羊肠线埋入后局部巨噬细胞和肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)表达的变化规律,初步探讨埋线后穴位局部初始应答机制。方法:SD大鼠随机分为空白组10只、羊肠线埋线组50只、埋线针空刺组50只,羊肠线埋线组、埋线针空刺组又随机分为干预后8 h,3、7、14、21 d组,每组10只。羊肠线埋线组进行“足三里”羊肠线埋线1次,埋线针空刺组进行“足三里”空刺1次。对应时间点采集各组“足三里”穴区组织,采用免疫荧光法检测穴区巨噬细胞CD68的表达,ELISA法检测“足三里”穴区组织TNF-α、IL-1β的含量。结果:与同时点空白组比较,羊肠线埋线组埋线后8 h,3、7、14、21 d穴区组织TNF-α、IL-1β含量及巨噬细胞CD68表达均升高(P<0.05);埋线针空刺组TNF-α含量空刺后8 h、3 d升高(P<0.05),IL-1β含量在空刺后8 h,3、7、14 d升高(P<0.05),巨噬细胞CD68表达在空刺后8 h升高(P<0.05)。与同时点埋线针空刺组比较,羊肠线埋线组3、7、14、21 d穴区组织IL-1β含量及8 h,3、7、14、21 d TNF-α含量、巨噬细胞CD68表达均升高(P<0.05)。结论:大鼠“足三里”实施羊肠线埋入后,局部巨噬细胞及相关细胞因子TNF-α、IL-1β含量发生变化,羊肠线为引起“足三里”穴区微环境变化的主要因素,其埋入后引起的局部免疫炎性动态变化可能参与了埋线后穴位局部初始应答。

Objective To observe the expression of local macrophages and related cytokines tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)after catgut implantation in“Zusanli”(ST 36)in rats,so as to explore its underlying mechanisms in inducing therapeutic effect.Methods A total of 110male SD rats were randomly divided into blank control group(n=10),catgut embedding(CE)group(n=50),and sham CE group(n=50).The CE and sham CE groups were randomly divided into 8h,3d,7d,14dand 21dsubgroups after the intervention(n=10in each time point group).Rats of the CE group were uniformly subjected into catgut embedding at ST36once,and those of the sham CE group received embedding needle puncture at ST36without catgut retention,and the blank control group was only grasped and fixed without other treatments.Tissues from the ST36area in each group were collected at the corresponding time points,and the expression of CD68in macrophages in the acupoint area was detected by immunofluorescence,the contents of TNF-αand IL-1βin the acupoint area were detected by ELISA.Results Following catgut embedment at ST36,the contents of TNF-αand IL-1β,and macrophage CD68expression level began to increase at 8h,peaked at 3d,and then gradually decreased at 7,14,and 21d,being still higher in the CE group than in the blank control group at 21d(P<0.05).Compared with the blank control group,the contents of TNF-αand IL-1β,and macrophage CD68expression were significantly increased at 8h,and 3,7,14and 21din the CE group(P<0.05).Following sham CE at ST36,the content of TNF-αat 8hand 3d,IL-1βat 8hand 3,7and 14d,and expression of CD68at 8hwere significantly increased in comparison with the blank control group(P<0.05).Comparison between the CE and sham CE groups showed that the contents of IL-1βat 3,7,14and 21d,and contents of TNF-α,CD68expression at 8h,and 3,7,14and 21dwere significantly higher in the CE group than in the sham CE group(P<0.05).Conclusion Catgut embedding at ST36can induce an increase levels of inflammatory cytokines TNF-α,IL-1βand macrophage CD68in the local microenvironment in rats,which may contribute to its functions in initiating therapeutic effect.