中国中西医结合急救杂志
中國中西醫結閤急救雜誌
중국중서의결합급구잡지
INTEGRATED TRADITIONAL CHINESE AND WESTERN MEDICINE IN PRACTICE OF CRITICAL CARE MEDICINE
2015年
1期
38-41
,共4页
于洪静%苑迅%姜玫%郝国华%李秋梅
于洪靜%苑迅%薑玫%郝國華%李鞦梅
우홍정%원신%강매%학국화%리추매
四妙勇安汤%平滑肌细胞%动脉粥样硬化%细胞外基质
四妙勇安湯%平滑肌細胞%動脈粥樣硬化%細胞外基質
사묘용안탕%평활기세포%동맥죽양경화%세포외기질
Simiao Yongan decoction%Smooth muscle cell%Atherosclerosis%Extracellular matrix
目的：观察四妙勇安汤药物血清对高胰岛素/高糖诱导的兔血管平滑肌细胞（VSMC）细胞外基质（ECM）分泌的影响及其可能的作用机制。方法采用血清药理学方法制备药物血清，高胰岛素/高糖诱导兔VSMC增殖。实验分为空白组〔10%胎牛血清（FBS）的正常DMEM〕、正常组（10%FBS的正常DMEM+正常血清）、模型组（10%FBS的高糖DMEM+100 U/L胰岛素+正常血清）、西药组（10%FBS的高糖DMEM+100 U/L胰岛素+辛伐他汀血清）、中药组（10%FBS的高糖DMEM+100 U/L胰岛素+四妙勇安汤血清）。用消化法检测细胞培养上清液中羟脯氨酸（Hyp）含量；逆转录-聚合酶链反应（RT-PCR）检测含药血清对兔VSMC的四型胶原蛋白（COL-Ⅳ）、基质金属蛋白酶-2（MMP-2）及其组织抑制因子（TIMP-1）的mRNA表达和对MMP-2/TIMP-1比值的影响。结果与模型组比较，中药组、西药组细胞培养上清液Hyp含量明显降低（均P＜0.05），且中药组明显低于西药组（mg/L：234.19±26.43比266.73±30.00，P＜0.05）。与正常组比较，模型组COL-Ⅳ、MMP-2的mRNA呈显著高表达（COL-ⅣmRNA：0.78±0.03比0.41±0.02，MMP-2 mRNA：0.80±0.12比0.41±0.02，均P＜0.05），TIMP-1 mRNA表达水平降低（0.35±0.04比0.79±0.07，P＜0.05），MMP-2/TIMP-1比值增加（2.30±0.35比0.52±0.05，P＜0.05）。与模型组比较，中药组、西药组COL-Ⅳ、MMP-2 mRNA表达水平均降低，TIMP-1 mRNA水平提高，MMP-2/TIMP-1比值有所下降（COL-ⅣmRNA：0.55±0.04、0.58±0.03比0.78±0.03，MMP-2 mRNA：0.62±0.05、0.67±0.08比0.80±0.12，TIMP-1 mRNA：0.56±0.02、0.60±0.01比0.35±0.04，MMP-2/TIMP-1：1.11±0.06、1.16±0.15比2.30±0.35，均P＜0.05），但中药组、西药组比较差异无统计学意义（均P＞0.05）。结论四妙勇安汤能够减少高胰岛素/高糖诱导的VSMC分泌ECM，其可能机制与减少Hyp合成，影响COL-Ⅳ、MMP-2、TIMP-1 mRNA表达水平，调整MMP-2/TIMP-1的平衡有关。
目的：觀察四妙勇安湯藥物血清對高胰島素/高糖誘導的兔血管平滑肌細胞（VSMC）細胞外基質（ECM）分泌的影響及其可能的作用機製。方法採用血清藥理學方法製備藥物血清，高胰島素/高糖誘導兔VSMC增殖。實驗分為空白組〔10%胎牛血清（FBS）的正常DMEM〕、正常組（10%FBS的正常DMEM+正常血清）、模型組（10%FBS的高糖DMEM+100 U/L胰島素+正常血清）、西藥組（10%FBS的高糖DMEM+100 U/L胰島素+辛伐他汀血清）、中藥組（10%FBS的高糖DMEM+100 U/L胰島素+四妙勇安湯血清）。用消化法檢測細胞培養上清液中羥脯氨痠（Hyp）含量；逆轉錄-聚閤酶鏈反應（RT-PCR）檢測含藥血清對兔VSMC的四型膠原蛋白（COL-Ⅳ）、基質金屬蛋白酶-2（MMP-2）及其組織抑製因子（TIMP-1）的mRNA錶達和對MMP-2/TIMP-1比值的影響。結果與模型組比較，中藥組、西藥組細胞培養上清液Hyp含量明顯降低（均P＜0.05），且中藥組明顯低于西藥組（mg/L：234.19±26.43比266.73±30.00，P＜0.05）。與正常組比較，模型組COL-Ⅳ、MMP-2的mRNA呈顯著高錶達（COL-ⅣmRNA：0.78±0.03比0.41±0.02，MMP-2 mRNA：0.80±0.12比0.41±0.02，均P＜0.05），TIMP-1 mRNA錶達水平降低（0.35±0.04比0.79±0.07，P＜0.05），MMP-2/TIMP-1比值增加（2.30±0.35比0.52±0.05，P＜0.05）。與模型組比較，中藥組、西藥組COL-Ⅳ、MMP-2 mRNA錶達水平均降低，TIMP-1 mRNA水平提高，MMP-2/TIMP-1比值有所下降（COL-ⅣmRNA：0.55±0.04、0.58±0.03比0.78±0.03，MMP-2 mRNA：0.62±0.05、0.67±0.08比0.80±0.12，TIMP-1 mRNA：0.56±0.02、0.60±0.01比0.35±0.04，MMP-2/TIMP-1：1.11±0.06、1.16±0.15比2.30±0.35，均P＜0.05），但中藥組、西藥組比較差異無統計學意義（均P＞0.05）。結論四妙勇安湯能夠減少高胰島素/高糖誘導的VSMC分泌ECM，其可能機製與減少Hyp閤成，影響COL-Ⅳ、MMP-2、TIMP-1 mRNA錶達水平，調整MMP-2/TIMP-1的平衡有關。
목적：관찰사묘용안탕약물혈청대고이도소/고당유도적토혈관평활기세포（VSMC）세포외기질（ECM）분비적영향급기가능적작용궤제。방법채용혈청약이학방법제비약물혈청，고이도소/고당유도토VSMC증식。실험분위공백조〔10%태우혈청（FBS）적정상DMEM〕、정상조（10%FBS적정상DMEM+정상혈청）、모형조（10%FBS적고당DMEM+100 U/L이도소+정상혈청）、서약조（10%FBS적고당DMEM+100 U/L이도소+신벌타정혈청）、중약조（10%FBS적고당DMEM+100 U/L이도소+사묘용안탕혈청）。용소화법검측세포배양상청액중간포안산（Hyp）함량；역전록-취합매련반응（RT-PCR）검측함약혈청대토VSMC적사형효원단백（COL-Ⅳ）、기질금속단백매-2（MMP-2）급기조직억제인자（TIMP-1）적mRNA표체화대MMP-2/TIMP-1비치적영향。결과여모형조비교，중약조、서약조세포배양상청액Hyp함량명현강저（균P＜0.05），차중약조명현저우서약조（mg/L：234.19±26.43비266.73±30.00，P＜0.05）。여정상조비교，모형조COL-Ⅳ、MMP-2적mRNA정현저고표체（COL-ⅣmRNA：0.78±0.03비0.41±0.02，MMP-2 mRNA：0.80±0.12비0.41±0.02，균P＜0.05），TIMP-1 mRNA표체수평강저（0.35±0.04비0.79±0.07，P＜0.05），MMP-2/TIMP-1비치증가（2.30±0.35비0.52±0.05，P＜0.05）。여모형조비교，중약조、서약조COL-Ⅳ、MMP-2 mRNA표체수평균강저，TIMP-1 mRNA수평제고，MMP-2/TIMP-1비치유소하강（COL-ⅣmRNA：0.55±0.04、0.58±0.03비0.78±0.03，MMP-2 mRNA：0.62±0.05、0.67±0.08비0.80±0.12，TIMP-1 mRNA：0.56±0.02、0.60±0.01비0.35±0.04，MMP-2/TIMP-1：1.11±0.06、1.16±0.15비2.30±0.35，균P＜0.05），단중약조、서약조비교차이무통계학의의（균P＞0.05）。결론사묘용안탕능구감소고이도소/고당유도적VSMC분비ECM，기가능궤제여감소Hyp합성，영향COL-Ⅳ、MMP-2、TIMP-1 mRNA표체수평，조정MMP-2/TIMP-1적평형유관。
Objective To observe the effects of serum containing Simiao Yongan decoction (SYD) on secretion of extracellular matrix (ECM) of rabbit vascular smooth muscle cells (VSMC) induced by high insulin/high glucose and its mechanism. Methods Serum containing SYD was prepared by serum pharmacological method, and the proliferation of rabbit VSMC was induced by high concentrations of glucose and insulin. The experiments were divided into following groups: blank group [normal glucose DMEM containing 10% fetal bovine serum (FBS)], normal group (normal glucose DMEM containing 10%FBS+normal serum), model group (high glucose DMEM containing 10%FBS+100 U/L insulin+normal serum), western medicine group (high glucose DMEM containing 10%FBS+100 U/L insulin+serum containing simvastatin), traditional Chinese medicine group (high glucose DMEM containing 10% FBS +100 U/L insulin+serum containing SYD). The content of hydroxy proline (Hyp) in cell culture supernatant was detected by digestion method. The reverse transcription-polymerase chain reaction (RT-PCR) was used to assay the effects of serum containing drug on the mRNA expressions of collagen-Ⅳ(COL-Ⅳ), matrix metalloproteinase-2 (MMP-2), tissue inhibitors metalloproteinase-1 (TIMP-1) and the MMP-2/TIMP-1 ratio of rabbit VSMC. Results Compared with the model group, the contents of Hyp in cell culture supernatant in traditional Chinese medicine group and western medicine group were significantly reduced (both P<0.05), and the content in traditional Chinese medicine group was much lower than that in western medicine group (mg/L:234.19±26.43 vs. 266.73±30.00, P<0.05). Compared with normal group, the mRNA expressions of COL-Ⅳand MMP-2 were significantly increased (COL-ⅣmRNA:0.78±0.03 vs. 0.41±0.02, MMP-2 mRNA: 0.80±0.12 vs. 0.41±0.02, both P < 0.05), the expression of TIMP-1 mRNA was decreased remarkably (0.35±0.04 vs. 0.79±0.07, P<0.05), and the ratio of MMP-2/TIMP-1 was increased obviously (2.30±0.35 vs. 0.52±0.05, P < 0.05) in model group. Compared with the model group, the mRNA expressions of COL-Ⅳand MMP-2 in traditional Chinese medicine group and western medicine group were decreased significantly, the expression of TIMP-1 mRNA was increased obviously, and the ratio of MMP-2/TIMP-1 was decreased (COL-ⅣmRNA: 0.55±0.04, 0.58±0.03 vs. 0.78±0.03, MMP-2 mRNA: 0.62±0.05, 0.67±0.08 vs. 0.80±0.12, TIMP-1 mRNA:0.56±0.02, 0.60±0.01 vs. 0.35±0.04, MMP-2/TIMP-1:1.11±0.06, 1.16±0.15 vs. 2.30±0.35, all P<0.05), but no statistically significant difference was found between traditional Chinese medicine group and western medicine group (all P > 0.05). Conclusion Serum containing SYD can reduce the secretion of ECM of the rabbit VSMC during the high state of glucose and insulin, and the mechanisms are possibly related to reducing the synthesis of Hyp, influencing the mRNA expression levels of COL-Ⅳ, MMP-2, TIMP-1, and adjusting the balance of MMP-2/TIMP-1.
