人参三七川芎提取物对增龄和高血压大鼠血管平滑肌细胞的影响

目的 观察和比较增龄与高血压对大鼠主动脉血管平滑肌细胞(vascular smooth muscle cells，VSMCs)的影响及人参、三七、川芎提取物的干预作用。方法 采用原代细胞培养的方法建立大鼠主动脉VSMCs模型，其中增龄实验分5组：青年对照组(Yon组)、老龄组(Old组)、老龄+普罗布考组(Old+Pro组)、老龄+中药低剂量组(Old+Low组)和老龄+中药高剂量组(Old+High组)；高血压实验分5组：Wistar-Kyoto对照组(WKY组)、自发性高血压大鼠(spontaneously hypertensive rat，SHR)组、SHR+缬沙坦组(SHR+Val组)、SHR+中药低剂量组(SHR+Low组)和SHR+中药高剂量组(SHR +High组)。采用MTT法检测细胞增殖，免疫细胞化学法检测基质金属蛋白酶-9(MMP-9)表达，RT-PCR和Western blot分别检测过氧化物酶增殖物激活受体γ(peroxisome proliferator activated receptor-γ，PPAR-γ)mRNA和蛋白表达。结果 与Yon组比较，Old组VSMCs增殖水平和MMP-9表达升高，PPAR-γ mRNA和蛋白表达减少，差异均有统计学意义(P<0.05)。与Old组比较，Old+Pro组、Old+High组和 Old+Low组VSMCs增殖水平和MMP-9表达明显降低，PPAR-γ 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(均P<0.05)。与WKY组比较，SHR组VSMCs增殖水平和MMP-9表达明显升高，PPAR-γ mRNA和蛋白表达明显减少(均P<0.05)；与SHR组比较，SHR+Val组、SHR +High组和SHR+Low组VSMCs增殖水平和MMP-9表达明显降低，PPAR-γ mRNA和蛋白表达增加(均P<0.05)。结论 增龄和高血压均可造成大鼠主动脉VSMCs过度增殖及相关细胞活性因子的表达改变，中药人参、三七、川芎提取物能够降低其增殖水平，减少负性细胞因子的表达以降低增龄和高血压对VSMCs的损害，延缓血管细胞的老化。

Effect of Extracts from Panax ginseng, Panax notoginseng, and Ligusticum Chuanxiong on Vascular Smooth Muscle Cells of Aging and Hypertension Rats

Objective 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. Methods 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+Probucal 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-γ (PPAR-γ) were detected using RT-PCR and Western blot respectively. Results Compared with the Yon group, the proliferation of VSMCs and the MMP-9 expression increased, the mRNA and protein expressions of PPAR-γ 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-γ obviously increased in the Old+Pro group, the Old+High group, and the Old+Low group (all P<0.05). The PPAR-γ 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-γ 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-γ obviously increased in the SHR+Val group, the SHR +High group, and the SHR+Low group (all P<0.05). Conclusions 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.