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目的:研究补阳还五汤含药血清对同型半胱氨酸(Hcy)致内皮细胞株(Eahy926)损伤的保护作用及其机制。方法:采用体外培养的内皮细胞株(Eahy926),测定不同的补阳还五汤含药血清对Hcy致内皮细胞活力的影响,检测细胞中超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)活力和丙二醛(MDA)含量;ELISA法检测可溶性细胞间黏附分子(sICAM-1)和肿瘤坏死因子-α(TNF-α)的浓度;荧光显微镜检测核转录因子-κB(NF-κB)的激活及Western blotting法测I-κBα蛋白表达。结果:10%,20%的补阳还五汤含药血清均能明显提高Hcy损伤的内皮细胞的存活率、SOD[分别为(15.87±0.46),(16.31±0.96)U·mL-1]活力,与损伤组(11.44±1.00)U·mL-1相比,P<0.01;降低LDH的释放和MDA[分别为(1.62±0.06),(1.47±0.18)nmol·L-1)的含量,与损伤组(1.89±0.25)nmol·L-1)相比,P<0.05;以及细胞培养液中sICAM-1[分别为(0.93±0.11),(1.00±0.14)μg·L-1]和TNF-α[分别为(63.71±2.64),(66.22±4.43)ng·L-1]浓度,与损伤组(2.32±0.23)μg·L-1,(125.67±2.69)ng·L-1)相比,P<0.05;同时抑制内皮细胞中NF-κB的激活,上调I-κBα蛋白表达量。结论:补阳还五汤含药血清对同型半胱氨酸致内皮细胞损伤具保护作用,其保护作用机制可能与抑制NF-κB的活化有关。
Objective: To study the protective effect of Buyang Huanwu Decoction-containing serum on homocysteine-induced endothelial cell injury (Eahy926) injury and its mechanism. Methods: Endothelial cells (Eahy926) cultured in vitro were used to determine the effects of different concentrations of Buyang Huanwu Decoction serum on the activity of Hcy induced endothelial cells. The levels of superoxide dismutase (SOD), lactate dehydrogenase (LDH) ) And malondialdehyde (MDA). The concentrations of soluble intercellular adhesion molecule (sICAM-1) and tumor necrosis factor-α (TNF-α) were detected by ELISA. The expressions of nuclear factor-κB ) Activation and Western blotting assay I-κBα protein expression. Results: The drug-containing serum of Buyang Huanwu Decoction (10%, 20%) significantly increased the survival rate of Hcy injured endothelial cells (SOD, 15.87 ± 0.46, 16.31 ± 0.96, (P <0.01). Compared with the injury group (11.44 ± 1.00) U · mL-1, P <0.01 decreased the release of LDH and the content of MDA (1.62 ± 0.06 and 1.47 ± 0.18 nmol·L-1, (1.93 ± 0.11) and (1.00 ± 0.14) μg · L-1, respectively, compared with those in the injury group (1.89 ± 0.25) nmol·L-1) (63.71 ± 2.64), (66.22 ± 4.43) ng · L-1] and TNF-α (2.32 ± 0.23) μg · L-1, (125.67 ± 2.69) ng · L -1 ), P <0.05; At the same time, it inhibited the activation of NF-κB in endothelial cells and up-regulated the expression of I-κBα. Conclusion: The serum of Buyang Huanwu Decoction contains protective effect of homocysteine on endothelial cell injury, and its protective mechanism may be related to the inhibition of the activation of NF-κB.