盐酸小檗碱对2型糖尿病大鼠血管平滑肌细胞增殖、迁移的影响
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摘要
目的观察盐酸小檗碱对2型糖尿病大鼠离体胸主动脉血管平滑肌细胞(VSMLs)增殖、迁移的影响,探究其机制。方法 SD大鼠高脂高糖饮食2个月,于1个月末腹腔注射链脲佐菌素25mg/kg制备2型糖尿病大鼠模型,组织块贴壁法培养胸主动脉VSMLs。CCK-8法观察不同浓度盐酸小檗碱对其增殖的影响、Transwell小室法观察其对迁移的影响、ELISA观察其对转化生长因子β1(TGF-β1)水平的影响。结果 100、50、25、12.5、6.25μg/mL的盐酸小檗碱均可抑制正常和糖尿病大鼠VSMLs的增殖,抑制强度与浓度呈正相关(P<0.01);各浓度盐酸小檗碱均可对正常和糖尿病大鼠VSMLs的迁移产生抑制作用,穿膜细胞数与浓度呈负相关(P<0.01);除6.25μg/mL的盐酸小檗碱外,其余浓度的盐酸小檗碱均可明显降低糖尿病大鼠VSMLs中TGF-β1水平(P<0.05)。结论盐酸小檗碱能抑制2型糖尿病大鼠离体胸主动脉VSMLs增殖、迁移,其机制可能与减少TGF-β1有关。
Objective To observe the effects of berberine hydrocloride on the proliferation and migration of vascular smooth muscle cells(VSMLs)of separated thoracic aorta in rats with type 2 diabetes mellitus(T2 DM)and to explore its mechanism.Methods The SD rats were fed with 2-month high fat and high sugar diet and conducted intraperitoneal injection of streptozotocin 25 mg/kg at the end of the first month for preparing the T2 DM rat model;the tissue block adherence method was used to culture vascular smooth muscle cells of thoracic aorta.The CCK-8 method was used to observe the effect of different concentrations of berberine hydrochloride on proliferation,and the effect of different concentrations of berberine on the migration was observed by Transwell chamber method.The effect of berberine hydrochloride on TGF-β1 level was observed by ELISA.Results 100,50,25,12.5,6.25μg/mL of berberine hydrochloride could inhibit the proliferation of VSMLs in normal and diabetic rats,and the inhibitory intensity was positively correlated with the concentration(P<0.01).Different concentrations of berberine hydrochloride all could produce the inhibiting effect on the migration ofbVSMLs in normal and diabetic rats.The number of transmembrane cells was negatively correlated with the concentration(P<0.01);excepting 6.25μg/mL of berberine hydrochloride,the other concentrations of berberine hydrochloride all could significantly decrease the TGF-β1 level in VMLs of diabetic rats(P<0.05).Conclusion Berberine hydrochloride can inhibit the proliferation and migration of VSMLs of thoracic aorta in T2 DM rats,and its mechanism may be related to the decrease of TGF-β1.
Objective To observe the effects of berberine hydrocloride on the proliferation and migration of vascular smooth muscle cells(VSMLs)of separated thoracic aorta in rats with type 2 diabetes mellitus(T2 DM)and to explore its mechanism.Methods The SD rats were fed with 2-month high fat and high sugar diet and conducted intraperitoneal injection of streptozotocin 25 mg/kg at the end of the first month for preparing the T2 DM rat model;the tissue block adherence method was used to culture vascular smooth muscle cells of thoracic aorta.The CCK-8 method was used to observe the effect of different concentrations of berberine hydrochloride on proliferation,and the effect of different concentrations of berberine on the migration was observed by Transwell chamber method.The effect of berberine hydrochloride on TGF-β1 level was observed by ELISA.Results 100,50,25,12.5,6.25μg/mL of berberine hydrochloride could inhibit the proliferation of VSMLs in normal and diabetic rats,and the inhibitory intensity was positively correlated with the concentration(P<0.01).Different concentrations of berberine hydrochloride all could produce the inhibiting effect on the migration ofbVSMLs in normal and diabetic rats.The number of transmembrane cells was negatively correlated with the concentration(P<0.01);excepting 6.25μg/mL of berberine hydrochloride,the other concentrations of berberine hydrochloride all could significantly decrease the TGF-β1 level in VMLs of diabetic rats(P<0.05).Conclusion Berberine hydrochloride can inhibit the proliferation and migration of VSMLs of thoracic aorta in T2 DM rats,and its mechanism may be related to the decrease of TGF-β1.
引文
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[6]彭麒朕,耿建国,邹大威,等.糖肾宁对高糖培养的小鼠系膜细胞TGF-β1和P38MAPK表达的影响[J].环球中医药,2017,10(3):265-269.
[7]陈纪言,傅国胜,傅向华,等.血管紧张素转换酶抑制剂在冠心病患者中应用中国专家共识[J].中国循环杂志,2016,31(5):420-425.
[8]朱清,萨仁高娃,岳欣,等.RAS拮抗剂调节血管平滑肌细胞增殖、凋亡因子表达[J].中国公共卫生,2012,28(5):633-636.
[9]LEE S,LIM H J,PARK H Y,et al.Berberine inhibits rat vascular smooth muscle cell proliferation and migration in vitro and improves neointima formation after balloon injury in vivo[J].Atherosclerosis,2006,186(1):29-37.
[10]李承德,王煜,曲敬蓉,等.黄芪多糖对糖尿病大鼠肾脏TGF-β1/Smads信号通路的影响[J].中国药理学通报,2018,34(4):512-516.
[11]徐云,陈雪辉,尹清风,等.TGF-β1/SGK1信号通路在糖尿病肾病中的作用[J].中国医师杂志,2017,19(10):1483-1487.
[2]张青,李琰,陈磊.黄连素对2型糖尿病及其并发症的治疗作用及相关机制研究进展[J].中国中药杂志,2015,40(9):1660-1665.
[3]贺娅莎,王彦.盐酸小檗碱对T2DM大鼠血内毒素的干预研究[J].中国医药指南,2017,15(23):26-27.
[4]梁宝今,蒙如庆,周卓东,等.盐酸小檗碱对大鼠主动脉内皮细胞增殖与凋亡的作用[J].右江医学,2016,44(6):621-624.
[5]乔进,陈敏,窦志华,等.转化生长因子β1/Smad信号转导途径在大黄酸保护糖尿病大鼠肾脏中的机制探讨[J].药学实践杂志,2017,35(5):402-406.
[6]彭麒朕,耿建国,邹大威,等.糖肾宁对高糖培养的小鼠系膜细胞TGF-β1和P38MAPK表达的影响[J].环球中医药,2017,10(3):265-269.
[7]陈纪言,傅国胜,傅向华,等.血管紧张素转换酶抑制剂在冠心病患者中应用中国专家共识[J].中国循环杂志,2016,31(5):420-425.
[8]朱清,萨仁高娃,岳欣,等.RAS拮抗剂调节血管平滑肌细胞增殖、凋亡因子表达[J].中国公共卫生,2012,28(5):633-636.
[9]LEE S,LIM H J,PARK H Y,et al.Berberine inhibits rat vascular smooth muscle cell proliferation and migration in vitro and improves neointima formation after balloon injury in vivo[J].Atherosclerosis,2006,186(1):29-37.
[10]李承德,王煜,曲敬蓉,等.黄芪多糖对糖尿病大鼠肾脏TGF-β1/Smads信号通路的影响[J].中国药理学通报,2018,34(4):512-516.
[11]徐云,陈雪辉,尹清风,等.TGF-β1/SGK1信号通路在糖尿病肾病中的作用[J].中国医师杂志,2017,19(10):1483-1487.