Lei Yan, Tao Li-li, Wang Guo-li
Medical Experiment Center, China Academy of Chinese Medical Sciences, Beijing.
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2012 Oct;32(10):1374-9.
To observe and compare the effects of aging and hypertension on rats' aortic vascular smooth muscle cells (VSMCs) and the effects of the extracts from Panax ginseng, Panax notoginseng and Ligusticum chuanxiong.
The rat aortic VSMCs model was established using the method of primary cell culture. Of them, the rats of the aging experiment were divided into 5 groups, i.e., the young control group (as Yon), the old group (as Old), the old + probucol group (as Old+Pro), the old +low dose extracts group (as Old+Pro), and the old+high dose extracts group (as Old+High). The rats of the hypertension experiment were divided into 5 groups, i.e., the Wistar-Kyoto control group (as WKY), the spontaneously hypertensive rat group (as SHR), the SHR +Valsartan group (as SHR+Val), the SHR+low dose extracts group (as SHR+Low), and the SHR+high dose extracts group (SHR+High). The proliferation of VSMCs was detected using MTT. The expression of MMP-9 was detected by immunocytochemical assay. The mRNA and protein expressions of peroxisome proliferator activated receptor-gamma (PPAR-gamma) were detected using RT-PCR and Western blot respectively.
Compared with the Yon group, the proliferation of VSMCs and the MMP-9 expression increased, the mRNA and protein expressions of PPAR-gamma decreased in the Old group, all showing statistical difference (P < 0.05). Compared with the Old group, the proliferation of VSMCs and the MMP-9 expression obviously decreased, the mRNA expression of PPAR-gamma obviously increased in the Old+Pro group, the Old+High group, and the Old+Low group (all P < 0.05). The PPAR-y protein expression obviously increased in the Old+Pro group and the Old+Low group (P < 0.05). Compared with the WKY group, the proliferation of VSMCs and the expression of MMP-9 obviously increased, the mRNA and protein expressions of PPAR-gamma obviously decreased in the SHR group (all P < 0.05). Compared with the SHR group, the proliferation of VSMCs and the expression of MMP-9 obviously decreased, the mRNA and protein expressions of PPAR-gamma obviously increased in the SHR+Val group, the SHR+High group, and the SHR + Low group (all P < 0.05).
Both aging and hypertension could result in excessive proliferation of rat aortic VSMCs and the expression changes of correlated cytoactive factors. The extracts from Panax ginseng, Panax notoginseng (Burk.) and Ligusticum chuanxiong can lower their proliferation levels and reduce the expressions of negative cytokines, thus reducing aging and hypertension induced injury of VSMCs and delaying angiocellular aging.
观察并比较衰老和高血压对大鼠主动脉血管平滑肌细胞(VSMCs)的影响以及人参、三七和川芎提取物的作用。
采用原代细胞培养法建立大鼠主动脉VSMCs模型。其中,衰老实验大鼠分为5组,即青年对照组(Yon)、老年组(Old)、老年+普罗布考组(Old+Pro)、老年+低剂量提取物组(Old+Low)和老年+高剂量提取物组(Old+High)。高血压实验大鼠分为5组,即Wistar-Kyoto对照组(WKY)、自发性高血压大鼠组(SHR)、SHR+缬沙坦组(SHR+Val)、SHR+低剂量提取物组(SHR+Low)和SHR+高剂量提取物组(SHR+High)。采用MTT法检测VSMCs的增殖情况。采用免疫细胞化学法检测MMP-9的表达。分别采用RT-PCR和Western blot法检测过氧化物酶体增殖物激活受体γ(PPAR-γ)的mRNA和蛋白表达。
与Yon组相比,Old组VSMCs的增殖和MMP-9表达增加,PPAR-γ的mRNA和蛋白表达降低,差异均有统计学意义(P<0.05)。与Old组相比,Old+Pro组、Old+High组和Old+Low组VSMCs的增殖和MMP-9表达明显降低,PPAR-γ的mRNA表达明显增加(均P<0.05)。Old+Pro组和Old+Low组PPAR-γ蛋白表达明显增加(P<0.05)。与WKY组相比,SHR组VSMCs的增殖和MMP-9表达明显增加,PPAR-γ的mRNA和蛋白表达明显降低(均P<0.05)。与SHR组相比,SHR+Val组、SHR+High组和SHR+Low组VSMCs的增殖和MMP-9表达明显降低,PPAR-γ的mRNA和蛋白表达明显增加(均P<0.05)。
衰老和高血压均可导致大鼠主动脉VSMCs过度增殖及相关细胞活性因子表达改变。人参、三七和川芎提取物可降低其增殖水平,减少负性细胞因子表达,从而减轻衰老和高血压所致的VSMCs损伤,延缓血管细胞衰老。