摘要
目的:观察丹参酮ⅡA衍生物-丹参酮ⅡA磺酸钠对血管紧张素Ⅱ诱导的心肌肥大及p-JNK,丝裂原活化蛋白激酶磷酸酶1表达的影响。方法:实验于2005-11/2006-03于华中科技大学同济医学院实验中心完成。取1d龄新生清洁级Wistar乳鼠10只,雌雄不拘,取心室肌组织分离培养新生大鼠心肌细胞,将细胞均匀地接种于6孔培养板,每孔2mL。第3天将培养的细胞分为5组,即对照组,血管紧张素Ⅱ组,丹参酮ⅡA磺酸钠2,10,50μmol/L组,分别给予相应药物。对照组给予生理盐水,其余各组均给予血管紧张素Ⅱ1μmol/L进行心肌细胞肥大诱导,在此基础上,丹参酮ⅡA磺酸钠2,10,50μmol/L组再分别给予相应浓度的丹参酮ⅡA磺酸钠,以上浓度为培养基内终浓度。采用考马斯亮蓝法测定心肌细胞蛋白含量;采用[3H]-亮氨酸掺入法测定蛋白合成速率作为心肌肥大指标;用Western-blot测定p-JNK,丝裂原活化蛋白激酶磷酸酶1表达。观察各组心肌细胞总蛋白质含量、[3H]-亮氨酸掺入测定结果、p-JNK,MKP-1蛋白表达。结果:①用刺激因素处理24h后,2,10,50μmol/L丹参酮ⅡA磺酸钠组总蛋白含量明显低于血管紧张素Ⅱ组[(65.38±1.26),(53.41±3.63),(48.42±2.61),(80.42±3.28)μg/孔,P<0.05～0.01]。②1μmol/L血管紧张素Ⅱ作用24h后,2,10,50μmol/L丹参酮ⅡA磺酸钠组心肌细胞合成速率显著低于血管紧张素Ⅱ组[(140.52±12.04)%,(120.58±8.72)%,(111.88±10.06)%,(163.04±11.38)%,P<0.01]。③1μmol/L血管紧张素Ⅱ作用24h后,2,10,50μmol/L丹参酮ⅡA磺酸钠组p-JNK蛋白表达显著低于血管紧张素Ⅱ组[(145.96±11.98)%,(133.04±6.54)%,(116.56±11.61)%,(167.04±12.72)%,P<0.05～0.01]。④丹参酮ⅡA磺酸钠(10μmol/L)显著上调丝裂原活化蛋白激酶磷酸酶1表达,在40min时达到高峰达(132.16±7.88)%,随后下降,在60,80min分别为(110.72±10.94)%,(104.32±9.55)%,(P<0.01)。结论:丹参酮ⅡA磺酸钠可以抑制血管紧张素Ⅱ诱导的心肌细胞肥大,机制可能与上调丝裂原活化蛋白激酶磷酸酶1表达,降低p-JNK表达有关。
AIM： To observe the effects of tanshinone Ⅱ A sulfonate （STS）, a derivate of tanshinone Ⅱ A, on angiotensin If-induced cardiomyocyte hypertrophy and expressions of phosphorylated JNK （p-JNK） and mitogen-activated protein kinase phosphatase （MKP-1）.
METHODS： The experiment was conducted in the Experimental Center of Tongji College, Huazhong University of Science and Technology between November 2005 to March 2006. Ten neonatal Wistar rats aged one-day old of clean grade and both sexes were selected to obtain the ventricular myocardium so as to isolate and culture the their myocardial cells. Cells were evenly inoculated into the 6-pore culture board with 2 mL in each pore. Cells were divided into 5 groups on the 3^rd day of culture： control group, Angiotensin-Ⅱ group, and 2, 10, 50 p, mol/L STS groups, and animals in each group were given corresponding drugs. Rats in the control group received normal saline, and rats in other groups were treated with saline; other groups were treated with Ang-Ⅱ （1μmoL/L） to induce cardiomyocyte hypertrophy, based on which rats in all STS groups were treated with different concentration of STS. The above-mentioned concentrations were the final concentrations of nutrient medium. The total protein in myocardial cells was determined by coomassie brilliant blue, and the protein synthesis rate was measured by [^3H]-Leucine incorporation. Expressions of p-JNK and MKP-1 were assessed by Western blot, and the total protein in my- ocardial cells, determination with [^3H]-Leucine incorporation and expressions of p-JNK and MKP-1 were observed in all groups.
RESULTS：①At 24 hours after treatment with stimulating factors, the total cellular protein of STS groups （2, 10, 50μmol/L） were obviously lower thah Ang-Ⅱ group [（65.38±1.26）, （53.41±3.63）, （48.42±2.61）, （80.42±3.28） p.g/pore, P 〈 0.05-0.01].②At 24 hours after treatment with Ang- Ⅱ, the protein synthesis rate of all STS groups （2, 10, 50μmol/L） were significantly, lower than the lower than the Ang-Ⅱ group [（140.52±12.04）%, （120.58±8.72）%, （111.88±10.06）%, （163.04±11.38）%, P 〈 0.01].③At 24 hours after treatment with 1μmol/L Ang-Ⅱ, the expressions of p-JNK in all STS groups were remarkably lower than the Ang-Ⅱ group[（145.96±11.98）%, （133.04±6.54）%, （116.56±11.61）%, （167.04±12.72）%,P 〈 0.05-0.01]. ④The expression of MKP-1 was obviously enhanced by STS （10μmol/L）, which reached to the peak （132.16±7.88）% at the 40^th minute, and then decreased to （110.72±10.94）% and （104.32±9.55）% respectively at the 60^th and 80^th minute （P 〈 0.01）.
CONCLUSION： STS can inhibit cardiomyoeyte hypertrophy induced by Ang-Ⅱ, and its mechanism may be related to the up-regulation of MKP-1 expression and the reduction p-JNK expression.
出处
《中国临床康复》
CAS
CSCD
北大核心
2006年第23期84-86,共3页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金资助项目(30500657)~~
