胡黄连苷Ⅱ降低M2型巨噬细胞的表达及对子宫内膜异位症大鼠盆腔黏连和内膜细胞凋亡的调节作用

Picroside Ⅱ reduces the expression of M2 macrophages and regulates pelvic adhesion and endometrial cell apoptosis in rats with endometriosis

目的探究胡黄连苷Ⅱ(PicrosideⅡ)对子宫内膜异位症(endometriosis,EMs)模型大鼠巨噬细胞的影响。方法将大鼠随机分为:对照组、EB组、Danazol组、低、中、高剂量PicrosideⅡ组,构建EMs模型并给予相应药物处理。根据各组大鼠盆腔黏连情况进行评分。通过TUNEL染色、RT-PCR、Western blot分别检测各组大鼠异位内膜细胞凋亡情况及Bax、Bcl-2的表达情况。其次,通过Western blot检测各组大鼠异位灶中VEGF、TGF-β、CD206、CD14、CD10表达情况。最后,通过流式细胞术分选各组大鼠异位灶中CD206+巨噬细胞。结果与对照组相比,EB组、Danazol组、低、中、高剂量PicrosideⅡ组大鼠黏连评分显著升高(P<0.05);与EB组相比,Danazol组、中、高剂量PicrosideⅡ组大鼠黏连评分显著降低(P<0.05)。Danazol组、中、高剂量PicrosideⅡ组中异位内膜细胞凋亡数量、Bax表达量较EB组显著增加(P<0.05),而Bcl-2表达量显著降低(P<0.05)。其次,与对照组相比,EB组、低、中剂量PicrosideⅡ组中VEGF、TGF-β、CD206、CD14、CD10表达量显著升高(P<0.05);与EB组相比,Danazol组、中、高剂量PicrosideⅡ组中VEGF、TGF-β、CD206、CD14、CD10表达量显著降低(P<0.05)。最后,与对照组相比,EB组、低、中剂量PicrosideⅡ组中CD206+巨噬细胞数量显著增多(P<0.05);与EB组相比,Danazol组、中、高剂量PicrosideⅡ组中CD206+巨噬细胞数量显著减少(P<0.05)。结论胡黄连苷Ⅱ抑制巨噬细胞极化促进细胞凋亡,并抑制模型大鼠异位灶的血管形成和纤维化。

To investigate the effect of Picroside Ⅱ on macrophages in rats with endometriosis(EMs), total of60 SD rats were recruited and randomly divided into control group, EB group, Danazol group, low, medium and high dose Picroside Ⅱ groups.EMs model was constructed and treated with corresponding drug.The pelvic adhesion of rats in each group was evaluated.TUNEL staining, RT-PCR and Western blot were used to detect the apoptosis of ectopic endometrial cells and the expression levels of Bax and Bcl-2 in each group;Western blot was also used to detect the expression levels of VEGF, TGF-beta, CD206, CD14 and CD10 in the ectopic foci of rats in each group;flow cytometry was used to isolate CD206+macrophages from ectopic foci of rats.Data showed that compared with the control group, adhesion scores of rats in EB group, Danazol group, low, medium and high dose Picroside Ⅱgroups were significantly increased(P<0.05).Compared with EB group, adhesion score of Danazol group, middle and high dose Picroside Ⅱ group were significantly decreased(P<0.05).The apoptosis and Bax expression of ectopic endometrial cells in the Danazol group and the medium and high dose Picroside Ⅱ groups were significantly increased compared with the EB group(P<0.05), while the expression of Bcl-2 was significantly decreased(P<0.05).Furthermore, compared with the control group, the expression levels of VEGF, TGF-β, CD206, CD14 and CD10 were significantly increased in the EB group and the Picroside Ⅱ group at low or medium doses(P<0.05);compared with EB group, the expression levels of VEGF, TGF-β, CD206, CD14 and CD10 were significantly decreased in the Danazol group and the medium and high dose Picroside Ⅱ groups(P<0.05).Finally, compared with the control group, CD206+macrophages were significantly increased in EB group, low and medium dose Picroside Ⅱ groups(P<0.05).While as compared with EB group, the number of CD206+macrophages in Danazol group, medium and high dose Picroside Ⅱ groups were significantly decreased(P<0.05).Taken together, Picroside Ⅱ inhibits the polarization and promotes apoptosis of macrophages, and inhibits the angiogenesis and fibrosis of ectopic foci in model rats.