Gong Xin, Jin Zhe, Lu Qiu-Dan, Lou Jiao-Ying
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2017 Apr;37(4):485-489.
Objective To observe the effect of Bushen Huoxue Recipe (BHR) on paracrine gene expression profiling of uterine natural killer cells (uNK cells) and uterine stromal cells. Methods Human stromal cells were extracted from proliferative phase endometrium of child-bearing age females, which were then divided into the blank group, the control group, and the BHR group. DMEM/F12 was added in cells of the BHR group to dilute into final concentration of 2 mg/mL herbal liquor. Equal volume of DMEM/ F12 was added to cells in the normal group and the control group. Cells in the control group and the BHR group were cultured for 24 h, with 20% serum-free DMEM plus 80% uNK cell secretion extracting solution added. Then they were cultured in 5% CΟ₂ at 37 °C for 6 h. Total RNAs were extracted after culture. The gene expression profile of stromal cells was detected using gene chip technology. At the same time mR- NA and protein expressions of chemokine (C-X-C motif) ligand 1 (CXCL1), intercellular cell adhesion molecule-1 (ICAM-1) , IL-8, and leukocyte inhibitor factor (LIF) were screened and detected using qRT- PCR and ELISA. Results Compared with the blank group, profiles of differentiated genes with 4-fold in- crease (a total of 63 genes) were basically agreeable in the control group and the BHR group. Compared with the control group, IL-15 receptor alpha (IL-15RA) was up-regulated by 1. 27 times, vascular endotheli- ai growth factor (VEGF) up-regulated by 1. 55 times, LIF up-regulated by 1. 45 times, IL-8 up-regulated by 1. 10 times, IL-11 up-regulated by 1. 23 times, transforming growth factor-β (TGF-β) up-regulated by 1. 40 times, epidermal growth factor (EGF) up-regulated by 1. 10 times, chemokine (C-C motif) ligand 8 (CCL8) up-regulated by 1.13 times, transporter 1 (TAP1 ) up-regulated by 1. 02 times, chemokine (C-X-C motif) receptor 2 (CXCR2) up-regulated by 1. 22 times, ICAM-1 up-regulated by 1. 15 times (P <0. 05) in the BHR group. Conclusion uNK paracrine played an important role in elevating endometrial receptivity and embry- o implantation, and BHR could improve and elevate the function of this paracrine system.
目的 观察补肾活血方（BHR）对子宫自然杀伤细胞（uNK细胞）和子宫基质细胞旁分泌基因表达谱的影响。方法 从育龄期女性增殖期子宫内膜中提取人基质细胞，分为空白组、对照组和BHR组。BHR组细胞加入DMEM/F12稀释成终浓度为2mg/mL的药液。正常组和对照组细胞加入等体积的DMEM/F12。对照组和BHR组细胞培养24h，加入20%无血清DMEM加80%uNK细胞分泌提取液。然后在37℃、5%CO₂条件下培养6h。培养后提取总RNA。采用基因芯片技术检测基质细胞的基因表达谱。同时采用qRT-PCR和ELISA筛选并检测趋化因子（C-X-C基序）配体1（CXCL1）、细胞间黏附分子-1（ICAM-1）、IL-8和白细胞抑制因子（LIF）的mRNA和蛋白表达。结果 与空白组相比，对照组和BHR组中差异基因表达上调4倍（共63个基因）的图谱基本一致。与对照组相比，BHR组中IL-15受体α（IL-15RA）上调1.27倍，血管内皮生长因子（VEGF）上调1.55倍，LIF上调1.45倍，IL-8上调1.10倍，IL-11上调1.23倍，转化生长因子-β（TGF-β）上调1.40倍，表皮生长因子（EGF）上调1.10倍，趋化因子（C-C基序）配体8（CCL8）上调1.13倍，转运体1（TAP1）上调1.02倍，趋化因子（C-X-C基序）受体2（CXCR2）上调1.22倍，ICAM-1上调1.15倍（P<0.05）。结论 uNK细胞旁分泌在提高子宫内膜容受性和胚胎着床方面起重要作用，BHR可改善并提升该旁分泌系统的功能。
