Zhao Yong-hua, Liu Yu-de, Liu Nai-wei
Faculty of Chinese Medicine, Macau University of Science and Technology, Macao.
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2010 Jan;30(1):68-71.
To observe the effects of Yinian Jiangya (YNJY) Decoction contained serum on cell proliferation of primary cultured endothelial cells of spontaneously hypertensive rat (SHR) and the peroxisome proliferator-activated receptor-gamma (PPAR-gamma) mRNA expression in the cells.
SD rats were fed with high-lipid diet and different doses (high, medium and low, containing crude drug of 5.2 g/mL, 2.6 g/mL, 1.3 g/mL respectively) of YNJY respectively and the serum contained different doses of YNJY (S-YNJY) was prepared by collecting the rats' serum after 20 days of feeding. The serum obtained from SD rats fed with normal diet, i.e. drug free serum (S-free) was taken for control. Primary cultured endothelial cells of SHR were treated respectively with S-YNJY in different doses (treated groups) and S-free (control group), the cell activity and the mRNA expression of PPAR-gamma in cells of all groups were detected by MTT and RT-PCR respectively at different time points: 2, 4, 8, 16 and 24 h after treatment.
MTT test showed that at time points of 4 and 8 h, the OD value raised in the treated groups with insignificant difference among them (P>0.05), but was higher than that in the control group (P<0.05); at 16 h, it increased but showed a smaller increment in the medium-dose treated group (P<0.05); at 24 h, it decreased in all groups, but the decrement in the high- and medium-dose treated groups was more remarkable (P<0.05); at 48 h, it decreased continuously, with the decrement more significant in the high-dose group than in the control group (P<0.05). RT-PCR detection showed that at 4 h, the expression of PPAR-gamma mRNA was not changed significantly in all groups (P>0.05); at 8 h, it was remarkably lower in high-dose group than in others (P<0.05); at 16 h, it was higher in the three treated groups than in the control group (P<0.05), with a highest level shown in the high-dose treated group (P<0.01); at 24 h, the expression decreased in all groups, but the level in the low-dose treated group was still higher than that in the control group (P<0.05).
YNJY Decoction shows a two-way regulation on endothelial cells proliferation, and which is negatively dose-dependent in the late stage. The regulation is perhaps relevant to the regulating of the PPAR-gamma mRNA expressions. The PPAR-gamma mRNA expression up-regulating and maintaining effects of YNJY Decoction may be one of the mechanisms for its vascular endothelial cell protection and blood pressure suppressing.
观察益年降压(YNJY)汤含药血清对自发性高血压大鼠(SHR)原代培养内皮细胞增殖及细胞中过氧化物酶体增殖物激活受体γ(PPAR-γ)mRNA表达的影响。
将SD大鼠分别喂以高脂饲料及不同剂量(高、中、低,分别含生药5.2 g/mL、2.6 g/mL、1.3 g/mL)的YNJY,喂养20天后采集大鼠血清,制备含不同剂量YNJY的血清(S-YNJY)。取喂以正常饲料的SD大鼠血清即不含药物血清(S-free)作为对照。将SHR原代培养的内皮细胞分别用不同剂量的S-YNJY(处理组)和S-free(对照组)处理,在处理后不同时间点(2、4、8、16和24小时)分别用MTT法和RT-PCR法检测各组细胞活性及细胞中PPAR-γ的mRNA表达。
MTT试验显示,在4和8小时时间点,处理组的OD值升高,各组间差异无统计学意义(P>0.05),但高于对照组(P<0.05);在16小时,OD值升高,但中剂量处理组升高幅度较小(P<0.05);在24小时,各组OD值均下降,但高剂量和中剂量处理组下降更显著(P<0.05);在48小时,OD值持续下降,高剂量组下降幅度比对照组更显著(P<0.05)。RT-PCR检测显示,在4小时,各组PPAR-γ mRNA表达无明显变化(P>0.05);在8小时,高剂量组PPAR-γ mRNA表达明显低于其他组(P<0.05);在16小时,三个处理组的PPAR-γ mRNA表达高于对照组(P<0.05),高剂量处理组表达水平最高(P<0.01);在24小时,各组表达均下降,但低剂量处理组水平仍高于对照组(P<0.05)。
YNJY汤对内皮细胞增殖具有双向调节作用,后期呈负性剂量依赖性。这种调节可能与PPAR-γ mRNA表达的调节有关。YNJY汤上调并维持PPAR-γ mRNA表达的作用可能是其保护血管内皮细胞和降低血压的机制之一。