不同刺法对单纯性子宫内膜增生模型小鼠巨噬细胞极化影响的比较研究OA北大核心CSTPCD
Comparative study on the effects of different acupuncture technique on macrophage polarization in a mouse model of simple endometrial hyperplasia
目的 观察不同刺法对单纯性子宫内膜增生模型小鼠子宫内膜影响的效应差异,并探讨其可能的作用机制.方法 32只雌性C57BL/6J小鼠按随机数字表法分为空白对照组、模型组、快针组、留针组,每组8只.采用双侧卵巢去势联合雌激素负荷法建立单纯性子宫内膜增生小鼠模型.快针组针刺小鼠双侧"隐白",刺之即出,隔日1次,共12次;留针组针刺小鼠双侧"隐白",每次留针15 min,隔日1次,共12次.干预结束后取材.HE染色法观察小鼠子宫组织形态学变化;酶联免疫吸附测定法检测血清中雌二醇水平;流式细胞术检测子宫组织M1、M2型巨噬细胞比例(M1/M2);免疫组织化学法检测子宫组织CD86、CD206表达;蛋白质印迹法检测子宫组织白细胞介素-13(IL-13)、γ 干扰素(IFN-γ)的表达.结果 模型组小鼠子宫内膜呈单纯性增生表现.与空白对照组比较,模型组子宫内膜增厚(P<0.01);血清雌二醇水平升高(P<0.01);子宫组织M1/M2下降(P<0.01);CD86阳性表达减少(P<0.01),CD206阳性表达增加(P<0.01);子宫组织IFN-γ蛋白表达降低(P<0.01)、IL-13蛋白表达增加(P<0.01).与模型组比较,快针组、留针组子宫内膜厚度降低(P<0.05);血清雌二醇水平降低(P<0.05);子宫组织M1/M2上升(P<0.01);CD206阳性表达、IL-13蛋白表达降低(P<0.01);CD86阳性表达、IFN-γ蛋白表达增加(P<0.01);与快针组比较,留针组IL-13蛋白表达增加(P<0.01).结论 快针和留针均可能是通过调控IFN-γ、IL-13的表达促使巨噬细胞由M2向M1型极化,抑制M2型巨噬细胞的促细胞增殖能力和组织修复能力,从而降低子宫内膜增生的程度.在调节IL-13的表达上快针优于留针.
Objective We aimed to observe the differences in the effects of different acupuncture technique on the endometrium of mice with simple endometrial hyperplasia model and to explore the potential mechanisms. Methods According to the random number tables,32 female C57BL/6J mice were divided into a blank control group,a model group,a quick needle group and a retaining needle group,with 8 mice in each group. A mouse model of simple endometrial hyperplasia was established using bilateral ovariectomy combined with estrogen loading. In the quick needle group,mice were punctured at the bilateral for "Yinbai"(SP1) points and withdrawn immediately,with the treatmeat performed once every other day for a total of 12 times. In the retaining needle group,mice were punctured at the bilateral "Yinbai"(SP1) points and the needles were retained for 15 min each time,with the treatment also performed once every other day for a total of 12 times. After the intervention,samples were collected. HE staining was used to observe morphological changes in the mouse uterine tissue;ELISA was used to detect serum estradiol level;flow cytometry was used to detect the ratio of M1 and M2 macrophages(M1/M2) and immunohistochemical method was used to measure the expression of CD86 and CD206 in uterine tissue;and Western blotting was used to detect the expression of interleukin-13 (IL-13) and interferon-γ(IFN-γ) in uterine tissue. Results The endometrium of mice in the model group showed simple hyperplasia. Compared with the blank control group,the endometrium of the model group was thickened (P<0.01);the level of estradiol in the serum was increased (P<0.01);M1/M2 in uterine tissues was decreased (P<0.01),the expression of CD86 was decreased (P<0.01),and the positive expression of CD206 was increased (P<0.01);and the level of IFN-γ protein expression in uterine tissues was decreased (P<0.01),and the expression of IL-13 protein was increased (P<0.01). Compared with the model group,the endometrial thicknesses of the quick needle group and the retaining needle group were reduced (P<0.05),the levels of estradiol in serum were reduced (P<0.05),M1/M2 in uterine tissues increased (P<0.01),and the reduction of CD206 positive expression,and IL-13 protein expression reduced (P<0.01);the level of CD86 positive expression,IFN-γ protein expression increased (P<0.01). Compared with the quick needle group,IL-13 protein expression increased in the retaining needle group (P<0.01).Conclusion Both quick needle and retaining needle may be through the regulation of the expression of IFN-γ and IL-13,thus prompting the polarization of macrophages from M2 to M1 type,inhibiting the pro-cell proliferative ability and tissue repair ability of M2 type macrophages,thus reducing the degree of endometrial hyperplasia,and the quick needle group was superior to the retaining needle group in regulating the expression of IL-13.
刘俊亚;李旭欣;张细芬;高渊博;佘延芬;李云峰;吕晶;郑文超;左广;王雪松;范玺胜;张俊茶;刘君
河北中医药大学针灸推拿学院 石家庄 050200河北中医药大学针灸推拿学院 石家庄 050200||河北省中医针灸优势病症国际联合研究中心河北中医药大学基础医学院
中医学
单纯性子宫内膜增生隐白巨噬细胞巨噬细胞极化快针留针小鼠
simple endometrial hyperplasiaYinbai(SP1)macrophagemacrophage polarizationquick needleretaining needlemice
《北京中医药大学学报》 2024 (009)
1203-1210 / 8
国家自然科学基金项目(No.81973755,No.81971434);河北中医药大学创新资助项目(No.XCXZZSS2023015)National Natural Science Foundation of China(Nos. 81973755 and 81971434)
评论