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目的:探讨山精胶囊对心肌缺血大鼠能量代谢、氧自由基及细胞凋亡相关基因的作用及其机制。方法:选取Wista大鼠采用冠状动脉结扎法,建立慢性心肌缺血模型。将筛选出来的大鼠随机分为山精胶囊低、中、高剂量组(283.5,850.5,1 701.0 mg·kg~(-1))和阳性药组(心安胶囊324.0 mg·kg~(-1)),假手术组和模型组灌胃给予同等剂量的0.9%的氯化钠注射液,给药10 d。紫外可见分光光度计检测血清及组织中乳酸(LD),游离脂肪酸(NEFA),丙二醛(MDA),超氧化物歧化酶(SOD),谷胱甘肽过氧化物酶(GSH-Px),一氧化氮(NO)及三磷酸腺苷酶(ATP)水平;蛋白质免疫印迹(Western blot)法及逆转录聚合酶链反应(RT-PCR)法检测心肌组织中核转录因子-κB(NF-κB),半胱氨酸蛋白酶-3(Caspase-3),B细胞淋巴瘤/白血病-2(Bcl-2)及Bcl-2相关X蛋白(Bax)蛋白表达。结果:与假手术组比较,模型组大鼠LD,NEFA,MDA明显升高(P<0.05),NO,Na~+-K~+-ATP,Ca~(2+)-ATP酶,SOD,GSH-Px明显降低(P<0.05),心肌NF-κB,Caspase-3,Bax mRNA及蛋白表达明显上调(P<0.05),Bcl-2 mRNA及蛋白表达明显下调(P<0.05);与模型组比较,山精胶囊能显著降低LD,NEFA,MDA(P<0.05),NO,Na~+-K~+-ATP,Ca~(2+)-ATP酶,SOD,GSH-Px显著升高(P<0.05),明显下调NF-κB,Caspase-3,Bax mRNA及蛋白表达(P<0.05),明显上调Bcl-2的mRNA及蛋白表达(P<0.05)。结论:山精胶囊可能通过调控NF-κB信号通路,提高心肌缺血大鼠清除氧自由基能力,改善心肌细胞能量代谢,调节凋亡相关蛋白和基因的表达,从而抑制了心肌细胞凋亡,减少缺血缺氧对心肌细胞的病理损害,起到保护心肌的作用。
Objective: To investigate the effect of Shanjing Capsule on energy metabolism, oxygen free radicals and apoptosis related genes in rats with myocardial ischemia. Methods: A rat model of chronic myocardial ischemia was established by coronary artery ligation in Wistar rats. The screened rats were randomly divided into two groups: shamian capsule low, medium and high dose group (283.5,850.5,1701.0 mg · kg -1) and positive group (Xinan capsule 324.0 mg · kg -1 ), Sham operation group and model group were given the same dose of 0.9% sodium chloride injection for 10 days. The contents of lactate (LD), free fatty acids (NEFA), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in serum and tissue were detected by UV- , Nitric oxide (NO) and adenosine triphosphatase (ATP) were detected by Western blotting and reverse transcription polymerase chain reaction (RT-PCR) Caspase-3, Bcl-2 and Bcl-2-related protein B (Bax) protein expression. Results: Compared with the sham operation group, the levels of NO, Na + -K + -ATP, Ca 2+ - ATPase, SOD, (P <0.05). The mRNA and protein expressions of NF-κB, Caspase-3 and Bax in myocardium were significantly increased (P <0.05) and the expressions of Bcl-2 mRNA and protein were significantly decreased Compared with sham group, shangjing capsule could significantly reduce the levels of LD, NEFA and MDA (P <0.05), NO, Na ~ + -K ~ + -ATP, Ca ~ (2 +) - ATPase, SOD and GSH-Px (P <0.05). The mRNA and protein expressions of NF-κB, Caspase-3 and Bax were significantly decreased (P <0.05), and the mRNA and protein expressions of Bcl-2 were significantly increased (P <0.05). Conclusion: Shanjing Capsule may inhibit cardiomyocyte apoptosis through regulating NF-κB signaling pathway, improving myocardial oxygen free radical scavenging capacity, improving myocardial energy metabolism, regulating apoptosis related protein and gene expression, Reduce ischemia and hypoxia on the pathological damage of myocardial cells, play a role in protecting the myocardium.