目的:研究脑心通对氧化低密度脂蛋白(ox-LDL)损伤血管内皮的保护作用及其分子机制.方法:在建立人脐静脉内皮细胞(HUVEC)ox-LDL(50 mg/L)损伤模型的基础上,以阿托伐他汀(1μmol/L)作为阳性对照,并给予0.2,0.4,0.6,0.8 g/kg不同剂量脑心通含药血清预先干预.采用硝酸还原酶法、放免法、分光光度计比色法、流式细胞技术等检测HUVEC NO,ET-1,LDH释放量,细胞内Caspase-3活性和早期凋亡率.结果:HUVEC损伤后,ox-LDL组(4.74±1.61)μmol/L与对照组(9.96±1.57)μmol/L相比较,NO的合成显著减少,而ET-1释放明显增加[(44.72±2.01)ng/L vs(21.42±2.84)ng/L,P＜0.01].与ox-LDL组(4.74±1.61)μmol/L相比较,ox-LDL+脑心通0.2,0.4,0.6,0.8 g/kg组[(8.33±1.78),(11.35±2.14),(14.80±2.40),(17.82±2.02)μmol/L,P＜0.05)]呈剂量依赖性,并可促进NO合成,但对ET-1释放无明显影响.而ox-LDL+阿托伐他汀组不仅可以促进NO合成,且能抑制ET-1释放(P＜0.01);ox-LDL可显著提高HUVEC的Caspase-3酶活性,促进LDH释放(P＜0.05),与ox-LDL组比较,ox-LDL+脑心通组对此呈剂量依赖性抑制作用[(193.00±16.81)μmoL/L vs(168.00±11.51),(135.00±11.18),(117.00±10.37),(99.00±9.62)μmol/L,P＜0.05],[(470.28±23.34)μmol/L vs 412.14±25.67),(311.37±28.34),(294.57±20.22),(276.49±25.32)μmol/L,P＜0.05],但阿托伐他汀对LDH释放无明显影响.脑心通、阿托伐他汀均能明显抑制ox-LDL诱导的内皮细胞凋亡(P＜0.01).结论:中药脑心通可促进内皮细胞NO释放,减轻内皮细胞的损伤与凋亡,是其保护血管内皮的可能机制.
Protective effect of Chinese herb NXT on endothelial cells
AIM: To investigate the protective effect of NaoXin-Tong(NXT) on vascular endothelial cell(HUVEC) from injury induced by ox-LDL and the possibly molecular mechanism. METHODS: ox-LDL of 50 mg/L was used and co-incubated with HUVECs to establish injured model of HUVEC. Atorvastatin (1 μmol/L)was used as positive control and different doses (0. 2,0.4,0.6 and 0.8 g) of NXT containing serum were used to intervene in advance. The contents of NO,ET-1 and LDH in the supernatant of HUVEC,the activity of intracellular Caspase-3 and the apoptosis of the HUVEC were examined by enzyme method,radio-immune method and Flow cytometry. RESULTS: Compared with those in normal control group,the contents of NO in the supernatant of ox-LDL-injured HUVECs significantly decreased [ (9.96 ±1. 57) μmol/L vs (4. 74 ± 1. 61) μmol/L) ],whereas the secretion of ET-1 significantly increased [ (21. 42 ± 2.84) ng/L vs (44.72±2.01) ng/L,P<0.05]. Compared with those in ox-LDL group,NXT promoted the contents of NO of ox-LDL-injured HUVECs in a dose dependent manner [ (8. 33 ± 1.78),(11.35 ±2. 14),(14.80±2.40),(17. 82 ±2. 02) μmol/L vs (4.74 ±1.61) μmol/L,P <0.05],but the secretion of ET-1 was not affected. Atorvastatin not only promoted the release of NO of HUVECs but also inhibited the secretion of ET-1 (P<0.01). The activity of intracellular Caspase-3 and the contents of LDH in the supernatant of ox-LDL-injured HUVEC significantly increased and NXT inhibited the effect in a dose dependent manner [ (193. 00 ± 16. 81) μmol/L vs (168. 00 ± 11. 51 ),(135.00±11.18),(117.00 ±10.37),(99.00±9.62) μmol/L,P<0.05],[(470.28±23.34)μmol/L vs (412.14 ±25. 67),(311.37 ±28. 34),(294. 57 ± 20. 22), (276. 49 ± 25. 32) μmol/L,P<0.05]. But atorvastatin had no effect on the release of LDH of HUVECs. NXT and atorvastatin both significantly inhibited the apoptosis of the ox-LDL-induced HUVECs (P < 0. 01). CONCLUSION: NXT promotes the release of NO and alleviates the injury and apoptosis of HUVECs induced by ox-LDL,which might be the underlying mechanism of the protective effect on HUVECs.
NETL
NSTL
万方数据
维普
