摘要
目的:探讨温经汤加味对子宫内膜异位症(EM)肾虚血瘀型大鼠的治疗作用及其机制研究。方法:从105只SPF级雌性未孕健康SD大鼠中随机选取10只作为空白组,余采用复合因素法构建肾虚血瘀型大鼠模型,造模成功后随机抽取10只为假手术组,仅开腹,不缝内膜,余采用自体内膜移植法构建EM肾虚血瘀型大鼠模型。从造模成功的56只大鼠中随机选取50只大鼠随机分为模型组、达那唑组(63 mg·kg^(-1))及温经汤加味低、中、高剂量组(5,10,20 g·kg^(-1)),每组10只,每日灌胃1次,连续4周。取各组内膜组织进行苏木素-伊红(HE)染色,观察组织病理变化;酶联免疫吸附测定(ELISA)检测各组血清上清液中白细胞介素-10(IL-10),白细胞介素-17(IL-17)含量;测量治疗前后各造模组异位灶长(D1),宽(D2),高(D3),并计算异位灶体积;免疫组化(IHC)检测各组大鼠内膜组织中Janus激酶2(JAK2),信号转导与转录活化因子3(STAT3),磷酸化STAT3(p-STAT3),血管内皮生长因子(VEGF),肿瘤坏死因子-α(TNF-α),血小板反应素-1(TSP-1)蛋白表达情况;蛋白免疫印迹法(Western blot)检测VEGF,TNF-α以及TSP-1蛋白表达情况。结果:镜下病理观察可见空白组大鼠子宫内膜腺体细胞排列整齐,腺体及间质细胞生长良好;与空白组比较,模型组子宫内膜结构完整,呈子宫腔状或环形封闭式结构,内有囊肿形成,上皮为立方状或柱状上皮,大部分上皮细胞有分泌现象,间质致密,基质呈少许纤维化,血管增生密集,并可见到少量腺体及炎细胞浸润。与空白组比较,模型组血清中IL-10含量显著降低(P<0.01),IL-17含量显著升高(P<0.01);模型组内膜组织中JAK2,STAT3,p-STAT3,VEGF,TNF-α蛋白表达明显升高(P<0.05),TSP-1蛋白表达明显降低(P<0.05)。与模型组比较,温经汤加味各剂量组大鼠血清中IL-10含量显著升高(P<0.01),IL-17含量显著降低(P<0.01),异位灶体积显著减小(P<0.01);温经汤加味高、中剂量组内膜
Objective:To investigate the therapeutic effect and mechanism of modified Wenjingtang on endometriosis(EM)rats with kidney deficiency and blood stasis. Method:The 10 from 105 SPF female healthy SD rats were randomly selected as the blank group. The rest constructed the rat model of kidney deficiency and blood stasis by compound factorial method. After the model was successfully established,10 rats were randomly selected as the sham operation group,with only laparotomy and no intima suture,and the remaining rats were established with EM kidney deficiency and blood stasis type by autologous intimal transplantation. Fifty rats which were randomly selected from 56 successful rats were treated with the modified Wenjingtang(5,10,20 g·kg^(-1))and danazol group(63 mg·kg^(-1)),1 time daily,for 4 weeks. The endometrial tissues of each group were stained with hematoxylin eosin(HE)to observe the histopathology. The levels of inflammatory factors interleukin-10(IL-10)and interleukin-17(IL-17)in serum supernatant were detected by enzyme linked immunosorbent assay(ELISA). Measuring the length(D1),width(D2)and height(D3)of the heterotopic foci in each group before and after treatment. Then calculating the volume of them. The expression of tyrosine kinase 2(JAK2),transcription factor 3(STAT3),phosphorylation transcription factor 3(p-STAT3),vascular endothelial growth factor(VEGF),tumor necrosis factor-α(TNF-α)and thrombospondin-1(TSP-1)were detected by immunohistochemistry(IHC). The expression of VEGF,TNF-α and TSP-1 was detected by Western blot.Result: Microscopic pathological observation showed that the endometrial glandular cells of the blank group were arranged in order,and the glandular and stromal cells grew well,compared with the blank group,the endometrial structure of the model group was complete,showing a cavity like or annular closed structure,with cyst formation,and the epithelium was cubic or columnar epithelium,most of the epithelial cells had secretion,the stroma was dense,and the matrix showed a little fibros
作者
孙瑞英
崔轶凡
曹娟
焦锐
SUN Rui-ying;CUI Yi-fan;CAO Juan;JIAO Rui(Second Clinical College,Shanxi University of Chinese Medicine,Taiyuan 030000,China;Fourth Clinical College,School of Basic Medicine,School of Nursing,Shanxi University of Chinese Medicine.Jinzhong 030619,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2021年第5期41-51,共11页
Chinese Journal of Experimental Traditional Medical Formulae
基金
山西省卫生和计划生育委员会科研课题(2017128)
山西中医药大学科技创新团队项目(2018TD-017)
山西省科技厅应用基础研究计划面上自然基金项目(201901D111341)。