摘要
目的研究不同浓度三七总皂苷对人脐静脉内皮细胞(HUVECs)增殖和血管生成能力的影响。方法体外培养HUVECs,分为空白对照组、三七总皂苷低浓度组(1 mg/L)、三七总皂苷中浓度组(20 mg/L)、三七总皂苷高浓度组(200 mg/L),各组给予相应浓度药物干预,分别孵育24 h、48 h、72 h后采用CCK-8法检测细胞活性;孵育48 h后采用EdU实验检测不同浓度三七总皂苷对HUVECs增殖能力的影响;孵育24 h后采用管腔形成实验检测不同浓度三七总皂苷对HUVECs血管生成能力的影响;孵育48 h后采用荧光定量PCR检测不同浓度三七总皂苷对HUVECs血管生成相关基因VEGF和eNOS mRNA表达的影响。结果CCK-8结果显示,与空白对照组比较,不同浓度三七总皂苷作用24 h、48 h、72 h对HUVEC细胞活性均无明显影响(P>0.05)。EdU结果显示,不同浓度三七总皂苷对HUVECs增殖能力无显著影响(P>0.05)。管腔形成实验结果表明,与空白对照组比较,三七总皂苷中浓度组(20 mg/L)可促进HUVECs血管生成(P<0.05),而三七总皂苷低浓度组(1 mg/L)、三七总皂苷高浓度组(200 mg/L)对HUVECs血管生成能力的影响与空白对照组比较差异无统计学意义(P>0.05)。荧光定量PCR结果显示,三七总皂苷中浓度组(20 mg/L)促进血管生成相关基因VEGF和eNOS mRNA表达,与空白对照组比较,差异有统计学意义(P<0.05),而与空白对照组比较,三七总皂苷低浓度组(1 mg/L)和三七总皂苷高浓度组(200 mg/L)对VEGF和eNOS mRNA表达无明显影响(P>0.05)。结论不同浓度三七总皂苷对HUVECs增殖能力无明显影响,但三七总皂苷中浓度组(20 mg/L)可促进HUVECs血管形成。
Objective To study the effects of Panax Notoginseng Saponins(PNS)at different concentrations on the proliferation and angiogenesis in human umbilical vein endothelial cells(HUVECs).Methods The HUVECs were cultured in vivo and divided into the blank control group,PNS group with low-dose(1 mg/L),PNS group with middle-dose(20 mg/L)and PNS group with high-dose(200 mg/L).Each group was treated with PNS at corresponding concentration.The cell activity was detected by CCK-8 assay after incubation for 24,48 and 72 hours respectively.The effect of PNS at different concentrations on the proliferation of HUVECs was detected by EdU test after incubation for 48 hours.The effect of PNS at different concentrations on the angiogenesis of HUVECs was detected by tube formation experiment after incubation for 24 hours.The effect of PNS at different concentrations on the mRNA expressions of VEGF and e NOS in HUVECs was detected by fluorescence quantitative PCR after incubation for 48 hours.Results The results of CCK-8 showed that compared with the blank control group,there was no significant effect on the activity of HUVECs after treated with different concentrations of PNS for 24,48 and 72 hours(P>0.05).EdU results showed that there was no significant effect on HUVECs proliferation after treated with different concentrations of PNS(P>0.05).The results of tube formation experiment showed that compared with the blank control group,the angiogenesis of HUVECs was promoted in the PNS group with middle-dose(20 mg/L)(P<0.05).However,there was no satistically significant difference in the angiogenesis of HUVECs between the blank control group and PNS group with low-dose(1 mg/L)or PNS group with high-dose(200 mg/L)(P>0.05).The results of fluorescence quantitative PCR showed that the mRNA expressions of VEGF and e NOS in PNS group with middle-dose(20 mg/L)were significantly higher than those in the blank control group,with statistically significant differences(P<0.05).Compared with the blank control group,the PNS group with low-dose(1 mg/L)or PNS group with highdose(200 mg/L)showed no significant effect on the mRNA expressions of VEGF and e NOS(P>0.05).Conclusion PNS at different concentrations shows no significant effect on the proliferation of HUVECs,but PNS at middle-dose(20 mg/L)could promote the angiogenesis of HUVECs.
作者
范德生
甄蕾
FAN Desheng;ZHEN Lei(Department of Pathology,Shanghai Baoshan District Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai 201999,China;Department of Periodontology,Shanghai Stomatological Hospital,Stomatological Hospital Affiliated to Fudan University,Shanghai 200001,China;Oral Biomedical Engineering Laboratory,Stomatological Hospital Affiliated to Fudan University,Shanghai 200001,China)
出处
《上海中医药杂志》
2020年第S01期142-146,共5页
Shanghai Journal of Traditional Chinese Medicine
基金
国家自然科学基金青年基金项目(81700981)
上海市自然科学基金项目(19ZR1438100)
上海市卫健委面上项目(201640103)
上海市宝山区科技创新专项资金项目(18-E-9)
上海市宝山区中西医结合医院国自然培育项目(GZRPYJJ-201804).
关键词
三七总皂苷
骨质疏松
人脐静脉内皮细胞
增殖
血管生成
Panax Notoginseng Saponins
osteoporosis
human umbilical vein endothelial cell
proliferation
angiogenesis
