葛根素延缓动脉粥样硬化进展的机制研究. (Chinese)

To clarify the therapeutic effect of Puerarin (Pur) on stability of atherosclerotic plaques and its underlying mechanism. High-fat-diet was used to induce atherosclerosis model, the mice were divided into Con group, ApoE -/- group and ApoE -/- +Pur group. The plaque area was measured by HE staining, the positive area of MMP2 in plaques were detected by immunofluorescence staining. In vitro, 50 滋g/mL ox-LDL was used to simulate atherosclerosis and Ad-sh-Sirt3 was used to inhibit Sirt3 expression. Cells were divided into Con group, ox-LDL group, ox-LDL + Pur group, ox-LDL + Pur + Ad-sh-Sirt3 group. Western-blot was used to detect the expression of Sirt3. The level of apoptosis in macrophages was detected by TUNEL assay. Compared with the Con group, the mice in the ApoE -/- group showed significant atherosclerotic plaques, the positive area of MMP2 in ApoE -/- mice were significantly higher than Con group (P<0.05). The plaque area and the positive area of MMP2 in ApoE -/- mice were significantly higher than ApoE -/- +Pur group (P<0.05). In cell level, western blot showed that compared with Con group, the expression of Sirt3 in ox-LDL group was significantly reduced (P<0.05). Treatment with Pur significantly increased the expression of Sirt3 compared with ox-LDL group (P<0.05). Furthermore, compared with the Con group, the level of macrophage apoptosis with ox-LDL administration was significantly increased (P<0.05), and Pur administration significantly reduced the apoptosis levels of cells compared with ox-LDL group (P<0.05). Pur effectively reduced the level of macrophage apoptosis and the infiltration of macrophages, increased the stability of atherosclerotic plaques by activating Sirt3 expression. [ABSTRACT FROM AUTHOR]

明确葛根素（Pur）对小鼠动脉粥样硬化斑块稳定性的治疗作用及其潜在机制。方法：用高脂饮食喂养成年小鼠诱导动 脉粥样硬化模型，将小鼠分为 Con 组，ApoE -/- 组，ApoE -/- + Pur 组。采用 HE 染色分别检测各组小鼠中动脉粥样硬化斑块面积，免疫 荧光染色检测斑块中 MMP2 的阳性区域面积。用 50 滋g/mL ox-LDL 干预巨噬细胞诱导动脉粥样硬化细胞模型，并用 Ad-sh-Sirt3 干扰 Sirt3 表达，将细胞分为 Con 组、ox-LDL 组、ox-LDL+Pur 组、ox-LDL+Pur+Ad-sh-Sirt3 组。Western-blot 检测 Sirt3 表达含量， TUNEL 法检测巨噬细胞凋亡。结果：动物水平，与 Con 组相比，ApoE -/- 组小鼠出现了显著的动脉粥样硬化斑块，ApoE -/- 组小鼠 MMP2 阳性区域面积显著高于 Con 组（P<0.05）；Pur 处理后，ApoE -/- + Pur 组动脉粥样硬化斑块面积明显低于 ApoE -/- 组（P<0.05）， MMP2 的阳性区域面积显著低于 ApoE -/- 组（P<0.05）。细胞水平，Western 结果显示，与对照组相比，ox-LDL 组 Sirt3 表达量显著降 低（P<0.05），ox-LDL+Pur 组 Sirt3 表达水平显著高于 ox-LDL 组（P<0.05）。相比于 Con 组，ox-LDL 组巨噬细胞凋亡水平显著升高 （P<0.05）；给予 Pur 处理后，相比于单纯 ox-LDL 组，ox-LDL+Pur 组巨噬细胞的凋亡水平显著降低（P<0.05）。Ad-sh-Sirt3 处理消除 了葛根素对于巨噬细胞凋亡的抑制作用（P<0.05）。结论：外源性 Pur 可能通过激活 Sirt3 表达，进一步降低巨噬细胞凋亡水平，减 少巨噬细胞的浸润，增加动脉粥样硬化斑块稳定性。 [ABSTRACT FROM AUTHOR]

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