目的 探討水飛薊賓對H₂O₂誘導(dǎo)狀態(tài)下H9C2心肌細胞氧化應(yīng)激損傷的保護作用。方法 將大鼠H9C2心肌細胞分為對照組、H₂O₂(200 μmol/L)干預(yù)組以及水飛薊賓(100、200 μmol/L)+H₂O₂(200 μmol/L)干預(yù)組,每組設(shè)6個復(fù)孔。各組經(jīng)過藥物干預(yù)6 h后,通過Giemsa染色法觀察細胞形態(tài)學(xué),通過MTT法測定細胞存活率、流式細胞術(shù)測定細胞凋亡率;測定細胞培養(yǎng)液中乳酸脫氫酶(LDH)、磷酸激酶(CK)、谷草轉(zhuǎn)氨酶(AST)活性和丙二醛(MDA)含量;測定細胞中超氧化物歧化酶(SOD)、過氧化氫酶(CAT)和谷胱甘肽過氧化物酶(GSH-Px)活性。結(jié)果 與對照組比較,H₂O₂干預(yù)組H9C2心肌細胞形態(tài)明顯異常、存活率顯著降低且凋亡率顯著升高,培養(yǎng)液中LDH、CK、AST活性和MDA含量均顯著升高,細胞中SOD、CAT、GSH-Px活性顯著降低,差異均具有統(tǒng)計學(xué)意義(P < 0.05)。與H₂O₂干預(yù)組比較,水飛薊賓(100、200 μmol/L)+H₂O₂(200 μmol/L)干預(yù)組培養(yǎng)液中AST、CK活性和MDA含量均顯著降低,差異具有統(tǒng)計學(xué)意義(P < 0.05);水飛薊賓(200 μmol/L)+H₂O₂(200 μmol/L)干預(yù)組H9C2心肌細胞形態(tài)明顯改善、存活率顯著升高、凋亡率顯著降低,培養(yǎng)液中LDH活性顯著降低,細胞中SOD、CAT、GSH-Px活性顯著升高,差異均具有統(tǒng)計學(xué)意義(P < 0.05)。結(jié)論 水飛薊賓能夠有效改善H₂O₂誘導(dǎo)狀態(tài)下H9C2心肌細胞形態(tài),提高其存活率并降低凋亡率,改善細胞中抗氧化酶活性、降低細胞損傷,提示水飛薊賓對H₂O₂誘導(dǎo)H9C2心肌細胞氧化應(yīng)激損傷具有劑量相關(guān)性的保護作用。

Objective To investigate the protective effects of silibinin on oxidative stress injury of H9C2 cardiomyocytes induced by H₂O₂. Methods H9C2 cardiomyocytes were randomly divided into control group, H₂O₂ (200 μmol/L) group, silibinin (100 μmol/L) +H₂O₂ (200 μmol/L) group, and silibinin (200 μmol/L) +H₂O₂ (200 μmol/L) group, and each group set six holes. After H₂O₂ stimulation for 6 h, the morphology changes were observed by microscope, and the survival rate and the apoptosis rate were detected by MTT method. The activity of LDH, CK, & AST, and the content of MDA in culture medium were detected, and the activities of SOD, CAT, and GSH-Px in cardiomyocytes were also determined. Results Compared with the control group, the morphology of H9C2 cardiomyocytes in H₂O₂ (200 μmol/L) group was abnormal, the survival rate was significantly decreased, and the apoptosis rate was significantly increased (P < 0.05). The activity of LDH, CK, & AST, and the content of MDA in culture medium were significantly increased (P < 0.05), the activity of SOD, CAT, and GSH-Px in cardiomyocytes were significantly decreased (P < 0.05). Compared with the H₂O₂ group, the activity of CK, AST, and the content of MDA in culture medium of silibinin (100, 200 μmol/L) +H₂O₂ (200 μmol/L) groups were significantly decreased (P < 0.05). The morphology of H9C2 cardiomyocytes in silibinin (200 μmol/L) +H₂O₂ (200 μmol/L) group was improved, the survival rate was significantly increased, and the apoptosis rate was significantly decreased (P < 0.05). The activity of LDH was significantly decreased (P < 0.05), and the activity of SOD, CAT, and GSH-Px in cardiomyocytes were significantly increased (P < 0.05). Conclusion Silibinin can effectively improve the morphology of H9C2 cardiomyocytes induced by H₂O₂, increase the survival rate and decrease the apoptosis rate, improve the activity of antioxidase, and depress the cell injury, which suggests that silibinin has dose-dependent protective effects against the oxidative stress of H9C2 cardiomyocytes induced by H₂O₂.

河北省衛(wèi)生廳重點科技研究計劃項目(20130359)