苦参碱对oxLDL 诱导的血管平滑肌细胞炎症反应及增殖凋亡的影响及分子机制研究

目的:探讨苦参碱对氧化型低密度脂蛋白(oxLDL)诱导的血管平滑肌细胞炎症反应及增殖凋亡的影响及分子机制。方法:oxLDL 处理人主动脉血管平滑肌细胞建立动脉粥样硬化模型。CCK-8 实验分析细胞活力和增殖。实时定量PCR(qRT-PCR)检测炎症因子白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、IL-10 和IL-13 的mRNA 水平。流式细胞术分析细胞凋亡。蛋白印迹检测增殖标记蛋白细胞增殖核抗原-67(Ki-67)和增殖细胞核抗原(PCNA),细胞凋亡标记蛋白B 细胞淋巴瘤-2(Bcl-2) 和Bcl-2 相关蛋白X(Bax),信号转导及转录激活子3(STAT3)和信号转导及转录激活子5(STAT5)的表达。结果:与对照组相比,造模组促炎因子IL-1β和TNF-α的mRNA 水平大大提高,抗炎因子IL-10 和IL-13 mRNA 水平则显著降低(P<0.01)。与造模组相比,模型加药组促炎因子IL-1β和TNF-α的mRNA 水平显著降低,抗炎因子IL-10 和IL-13 的mRNA 水平明显升高(P<0.05)。造模组细胞增殖倍数和细胞凋亡率明显高于对照组,模型加药组细胞增殖倍数和细胞凋亡率明显低于造模组(P<0.05)。与对照组相比,造模组Ki-67、PCNA 和Bax 的表达显著升高,Bcl-2 显著降低(P<0.01)。与造模组相比,模型加药组Ki-67、PCNA 和Bax 的表达明显减少,Bcl-2 表达明显增多(P<0.05)。造模组p-STAT3 和p-STAT5 相对蛋白表达量显著高于对照组(P<0.01)。模型加药组p-STAT3 和p-STAT5 相对蛋白表达量明显低于造模组(P<0.05)。与造模组相比,模型加药组和模型+Ruxolitinib 组Ki-67、PCNA 和Bax 的表达明显减少,Bcl-2 表达明显增多(P<0.05);模型加药+Ruxolitinib 组Ki-67、PCNA 和Bax 的表达显著减少,Bcl-2 表达显著增多(P <0.01)。与造模组相比,模型加药组和模型+Ruxolitinib 组及模型加药+Ruxolitinib 组IL-1β和TNF-α的mRNA 水平都明显降低,IL-10 和IL-13 的mRNA 水平都明显升高(P<0.05)。结论:苦参碱可通过抑制JAK/ STAT3 通路的活化起到抗炎和抑制血管平滑肌细胞增殖和凋亡的作用。

Ginsenoside Rb1 ameliorates abnormal glucolipid metabolism of liver through inhibition of JNK signal pathway in diabetic rats

Objective:To explore effects and mechanism on glucolipid metabolism of liver of ginsenoside Rb1 in diabetic rats.Methods: Diabetic rat model was induced by streptozotocin (STZ).Hepatic pathological changes were observed by hematoxylin-eosin (HE)staining.Apoptosis was analyzed through Terminal deoxynucleotidyl transferase-mediated dUTP nick labeling (TUNEL) staining.Blood glucose was determined with glucometer.The level of insulin and C peptide was tested by radioimmunoassay.Total cholesterol, low density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglyceride was detected by full automatic biochemical analyzer.The expression of JNK signal pathway related proteins JNK1 and c-Jun was measured by Western blot.The expression of inflammatory factor IL-6,IL-1βand TNF-αwas detected by immunohistochemistry.Results: Liver injury was ameliorated by ginsenoside Rb1 in diabetic rats.Compared with control group,the level of blood glucose in STZ-induced diabetic rat group was increased with attenuated level of insulin and C peptide (P<0.05).Compared with STZ-induced diabetic rat group,the level of blood glucose in STZ+ginsenoside Rb1 group was decreased with enhancive level of insulin and C peptide (P<0.05).Compared with control group,the level of total cholesterol,triglyceride and low density lipoprotein cholesterol in STZ-induced diabetic rat group was elevated with attenuated level of high-density lipoprotein cholesterol (P<0.05).Compared with STZ-induced diabetic rat group,the level of total cholesterol,triglyceride and low density lipoprotein cholesterol in STZ+ginsenoside Rb1 group declined with elevated level of high-density lipoprotein cholesterol (P<0.05). The relative expression of JNK1 and c-Jun in STZ-induced diabetic rat group was higher than control group (P<0.05).The relative expression of JNK1 and c-Jun in STZ+ginsenoside Rb1 group was lower than STZ-induced diabetic rat group (P<0.05). Compared with control group,the expression of IL-6,IL-1βand TNF-αin STZ-induced diabetic rat group was enhanced (P<0.05).The expression of IL-6,IL-1βand TNF-α in STZ+ginsenoside Rb1 group was lower than STZ-induced diabetic rat group (P<0.05).Conclusion: Ginsenoside Rb1 ameliorates liver injury,abnormal glucolipid metabolism and inflammatory response of liver through inhibition of JNK signal pathway in diabetic rats.