含活血通络方血清对高糖诱导人视网膜血管内皮细胞增殖作用的研究

目的 探究含活血通络方血清调控IncRNA MEG3对高糖诱导人视网膜血管内皮细胞（human retinal microvascular endothelialcells,hRMECs）增殖及凋亡的影响作用。方法 选取20只SD大鼠，采用随机数字表法对10只大鼠灌胃活血通络方，10只灌胃等量生理盐水。灌胃结束1h后制备相应血清。分别采用5mmol/L葡萄糖或30mmol/L的D-甘露醇、葡萄糖溶液对hRMECs细胞进行诱导24h。按照诱导情况将细胞分为正常对照组（5mmol/L葡萄糖，n=6）、等渗组（30mmol/LD-甘露醇，n=6）、高糖组（30mmol/L葡萄糖，n=6）、高糖+2.5%含药血清组（n=6）、高糖+7.5%含药血清组（n=6）、高糖+10%含药血清组（n=6），相应血清干预24h。采用CCK-8法、酶联免疫吸附法检测细胞增殖情况及丙二醛（malondialdehyde,MDA）、谷胱甘肽过氧化物酶（glutathioneperoxidase,GSH-Px）含量；采用实时荧光定量聚合酶链反应（quantitative real-time fluorescence...

Study on the proliferation of human retinal microvascular endothelial cells induced by high glucose in Huoxue Tongluo formula containing serum

Objective To investigate the effect of Huoxue Tongluo formula regulates IncRNA MEG3 on the proliferation and apoptosis of human retinal microvascular endothelial cells (hRMECs) induced by high glucose. Methods Twenty SD rats were selected and 10 rats were gavaged with Huoxue Tongluo formula and 10 rats were gavaged with equal amount of saline according to random number table method. The corresponding serum was prepared 1h after the end of gavage. The hRMECs cells were induced with 5mmol/L glucose or 30mmol/L D-mannitol, glucose solution for 24h. Cells were divided according to induction into normal control (5mmol/L glucose, n=6), isotonic (30mmol/L D-mannitol, n=6), high glucose (30mmol/L glucose, n=6), high glucose+2.5% drug-containing serum (n=6), high glucose+7.5% drug-containing serum (n=6) and high glucose+10% drug-containing serum (n=6) groups with the corresponding serum intervention for 24h. Cell proliferation and malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) levels were measured by CCK-8 and ELISA. Cellular IncRNA-MEG3 mRNA expression was measured by quantitative real-time fluorescence polymerase chain reaction (qRT-PCR) and apoptosis was measured by flow cytometry. Protein immunoblotting was used to detect NOX4, VAV2, vascular epidermal growth factor receptor (VEGFR), phosphoinositide 3-kinase (PI3K), p-PI3K, protein kinase B (AKT), p-AKT, mammalian target of rapamycin (mTOR), p-mTOR protein expression. Results Compared with the normal control group, the high glucose group showed decreased cell proliferation rate, increased MDA content, decreased GSH-Px content, increased apoptosis, decreased IncRNA-MEG3 mRNA expression, and increased p-PI3K/PI3K, p-AKT/AKT, p-mTOR/mTOR, NOX4, VAV2, and VEGFR protein expression, all with statistically significant differences (P<0.05). Compared with the high glucose group, the cell proliferation rate increased, MDA content decreased, GSH-Px content increased and IncRNA-MEG3 expression increased in the high glucose +7.5% drug-containing serum group and the high glucose+10% drug-containing serum group, all with statistically significant differences (P<0.05). Apoptosis was decreased in the high glucose+2.5% drug-containing serum group, the high glucose+7.5% drug-containing serum group and the high glucose+10% drug-containing serum group, and p-PI3K/PI3K, p-AKT/AKT, p-mTOR/mTOR, NOX4, VAV2, VEGFR protein expression were decreased, all differences were statistically significant (P<0.05). Conclusion Huoxue Tongluo formula mediates IncRNA-MEG3-regulated protein expression to reduce apoptosis in hRMECs induced by high glucose.