Bushen Huoxue decoction(补肾活血汤)improves the reproduction of endometriosis-associated infertility by regulating Homeobox A10 and αlpha(v)beta(3)integrin expression

OBJECTIVE: To investigate the mechanism of Bushen Huoxue decoction( 补肾活血汤, BSHXD) to treat endometriosis-induced infertility. MEDHODS: The main compounds of BSHXD were determined by high performance liquid chromatographymass spectrometry(HPLC-MS/MS). The effect of BSHXD on Homeobox A10(HOXA10) and alpha(v)beta(3)(αvβ3) integrin expression of Ishikawa cells, mouse model, and endometriosis-associated infertility women was evaluated by using Western blot analysis, immunohistochemistry and Real-Time quantitative polymerase chain reaction(RTq PCR). The efficacy of BSHXD on embryo attachment were examined by using the Be Wo spheroid and mouse embryo attachment assay. HOXA10 concentration in uterine flushing fluid of endometriosis-associated infertility women treated with BSHXD was measured by EnzymeLinked immunosorbent assay(ELISA).RESULTS: BSHXD improved Be Wo spheroid and mice blastocysts attachment to Ishikawa cells and increased embryo implantation rates in mice and pregnancy rates in women with endometriosis-associated infertility. BSHXD enhanced HOXA10 and αvβ3 integrin expression in Ishikawa cell, endometriosis mouse model, and endometriosis-associated infertility women, which potentially improved endometrial receptivity. CONCLUSIONS: BSHXD could improve endometrial receptivity of endometriosis-associated infertility in a dosedependent manner by regulating HOXA10 and αvβ3 integrin expression.

骨桥蛋白-整合素ɑνβ3配体受体反应对人膝骨关节炎软骨细胞透明质酸表达水平的影响

目的通过干预体外培养人膝骨关节炎软骨细胞中的骨桥蛋白(OPN)-整合素aB3配体受体反应,观察其对较骨细胞透明质酸(HA)表达水平的影响。方法从人膝骨关节炎软骨组织标本中获取并体外培养软骨细胞,实验分为三组:对照组加入1μg/ml重组人骨桥蛋白(rhOPN)干预;整合素ayβ3阻断组先采用整合素avB3阻断剂CRGDSP peptide(100μ/ml)干预软骨细胞1h以阻断OPN-整合素avβ3的结合后再加人1μ/ml thOPN进行干预23 h,整合素avβ3同型组先采用整合素av33同型剂GRGESP peptide 100μ/ml干预软骨细胞1h后再加入1μg/ml rhOPN进行干预23 h.采用实时荧光定量PCR(qPCR)法检测各组透明质酸合成酶HASI,HAS2和HAS3 mRNA表达,采用ELISA法检测HA水平。结果显微镜下观察膝骨关节炎软骨细胞与正常软骨细胞培养肩的形态,正常软骨细胞呈多角形,而膝骨关节炎软骨细胞伪足增多,形态多不规则。另外,与对照组(1.01±0.10)相比,整合素avB3阻断组HASI,HAS2,HAS3 mRNA表达水平[(0.70±0.01)、(0.68±0.05)、(0.55±0.03)]显著降低,差异有统计学意义(P<0.05);而整合素avβ3同型组HASI HAS2,HA53 mlRNA表达水平[(1.09±0.14)、(0.95±0.06).(1.04±0.08)]与对照组比较差异无统计学意义(P>0.05)。与对照组[(884.0±7.6)pg/ml]比较,整合素avβ3阻断组HA水平[(578.4±13.8)μg/ml]显著降低,差异有统计学意义(P<0.05);而整合素apβ3同型组HA水平[(882.1±10.1)μg/ml]与对照组比较差异无统计学意义(P>0.05)。结论OPN可以通过与整合素avB3配体受体反应刺激软骨细胞HAS分泌进而诱导HA表达。