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Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (1): 76-84.doi: 10.3736/jcim20120112

• Original Experimental Research • Previous Articles     Next Articles

Effects of extracts from Panax ginseng, Panax notoginseng and Ligusticum chuanxiong on expression of β-galactosidase and signal pathway p16-cyclin D/CDK-Rb in vascular smooth muscle cells

Tao Li-li1,Lei Yan2()   

  1. 1. Cardiovascular Center, Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing 100091, China
    2. Medical Experiment Center, China Academy of Chinese Medical Sciences, Beijing 100700, China
  • Received:2011-08-04 Accepted:2011-09-12 Online:2012-01-20 Published:2018-09-21
  • Contact: Lei Yan E-mail:leiy999@163.com

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OBJECTIVE: To observe the pathological changes of vascular aging in spontaneously hypertensive rats (SHRs), and the effects of extracts from Panax ginseng, Panax notoginseng (Burk.) and Ligusticum chuanxiong on vascular aging.METHODS: Vascular smooth muscle cells (VSMCs) isolated from thoracic aorta of Wistar-Kyoto (WKY) rats and SHRs were primarily cultured and divided into WKY group, SHR group, valsartan group, low-dose extract group and high-dose extract group. Aging situation of VSMCs was detected by senescence-associated β-galactosidase (SA-β-gal) staining; the cell cycle was analyzed by flow cytometry; mRNA and protein expressions of p16, cyclin D1, cyclin-dependent kinase 4 (CDK4) and retinoblastoma protein (Rb) were detected by real-time fluorescence quantitative polymerase chain reaction (PCR) and Western blotting.RESULTS: SA-β-gal stain showed that compared with the WKY group, number of SA-β-gal-positive cells in the SHR group was increased significantly (P<0.01); extracts from Panax ginseng, Panax notoginseng (Burk.) and Ligusticum chuanxiong reduced the number of SA-β-gal-positive cells in the SHR group (P<0.01). Flow cytometry analysis showed that the number of VSMCs in G1 phase was reduced and in S phase was increased significantly (P<0.05) in the SHR group compared with the WKY group, and the number of VSMCs in G1 phase was increased and in S phase was reduced significantly in the extracts groups (P<0.05). PCR result showed that compared with the WKY group, the mRNA expressions of p16 and Rb in the SHR group were reduced and those of cyclin D1 and CDK4 were increased significantly, the mRNA expressions of p16 and Rb in the extracts group were increased and mRNA expressions of cyclin D1 and CDK4 were reduced significantly. Western blotting result showed that compared with the WKY group, the protein expression of p16 in the SHR group was reduced and the protein expressions of cyclin D1, CDK4 and phospho-Rb were increased significantly (P<0.05), while the protein expression of p16 in the extracts group was increased and the protein expressions of cyclin D1, CDK4 and phospho-Rb were reduced significantly (P<0.05).CONCLUSION: The extracts from Panax ginseng, Panax notoginseng (Burk.) and Ligusticum chuanxiong can delay vascular aging of SHRs, which works by p16-cyclin D/CDK-RB pathways to inhibit VSMC proliferation.

Key words: hypertension, vascular aging, plant extracts, senescence-associated β-galactosidase;, gene, p16, cyclin D1, cyclin-dependent kinase 4, retinoblastoma protein

Figure 1

Growth morphology of VSMCs and identificationA: Cells obtained from aorta of rats had a spindle shape (Inverted microscopy, ×200); B: Cells displayed a hill-and-valley pattern typical for VSMCs (Inverted microscopy, ×100); C: Percentage of α-actin-positive smooth muscle cells was more than 95% (Inverted microscopy, ×400). VSMCs: vascular smooth muscle cells."

Table 1

SA-β-gal-positive VSMCs in different groups ($\bar{x}$±s, %)"

Group n SA-β-gal-positive VSMCs
Wistar-Kyoto 3 11.14±2.21
SHR 3 32.89±3.14**
Valsartan 3 15.96±1.59△△
Low-dose extract 3 24.36±2.00△△
High-dose extract 3 20.78±1.86△△

Figure 2

SA-β-gal-positive VSMCs in different groups (×400) A: Wistar-Kyoto group; B: Spontaneously hypertensive rat group; C: Valsartan group; D: Low-dose extract (20 mg/L) group; E: High-dose extract (40 mg/L) group. SA-β-gal: senescence-associated β-galactosidase; VSMC: vascular smooth muscle cell."

Table 2

Cell cycle analysis of VSMCs ($\bar{x}$±s, %)"

Group n G1 phase G2 phase S phase
Wistar-Kyoto 3 91.23±1.10 6.75±0.83 2.02±1.21
SHR 3 67.40±0.85* 17.43±0.34* 15.17±0.99*
Valsartan 3 87.90±8.49△ 10.53±0.96△ 2.71±1.57△
Low-dose extract 3 81.67±0.81△ 15.12±0.17 3.22±0.91△
High-dose extract 3 85.43±0.96△ 7.53±0.56△ 7.03±1.52△

Figure 3

Cell cycle analysis of VSMCsA: Wistar-Kyoto group; B: Spontaneously hypertensive rat group; C: Valsartan group; D: Low-dose extract (20 mg/L) group; E: High-dose extract (40 mg/L) group. VSMC: voscular smooth muscle cell."

Table 3

Expressions of p16, cyclin D1, CDK4 and Rb mRNAs in VSMCs ($\bar{x}$±s)"

Group n p16 Cyclin D1 CDK4 Rb
Wistar-Kyoto 3 3.31±0.06 0.27±0.02 1.01±0.02 2.87±0.04
SHR 3 0.79±0.38* 1.25±0.24* 3.14±0.19* 0.57±0.49*
Valsartan 3 2.52±0.12△ 0.45±0.03△ 1.24±0.01△ 2.30±0.17△
Low-dose extract 3 1.39±0.11△ 0.77±0.10△ 3.01±0.30 1.17±0.32△
High-dose extract 3 1.76±0.21△ 0.63±0.15△ 2.70±0.09 1.74±0.05△

Table 4

Expressions of p16, cyclinD1 and CDK4 proteins and pRb in VSMCs ($\bar{x}$±s)"

Group n p16 Cyclin D1 CDK4 pRb
Wistar-Kyoto 3 1.19±0.31 0.82±0.06 1.37±0.13 0.50±0.14
SHR 3 0.51±0.06* 2.17±0.35* 2.17±0.27* 1.34±0.08*
Valsartan 3 0.89±0.23△ 0.93±0.07△ 0.95±0.20△ 0.56±0.01△
Low-dose extract 3 1.04±0.14△ 1.52±0.32△ 1.81±0.13△ 1.19±0.21△
High-dose extract 3 1.03±0.09△ 1.47±0.15△ 2.00±0.35 0.86±0.07△

Figure 4

Expressions of p16, cyclinD1, CDK4 proteins and pRb in VSMCsA: Wistar-Kyoto group; B: SHR group; C: Valsartan group; D: Low-dose extract (20 mg/L) group; E: High-dose extract (40 mg/L) group. SHR: spontaneously hypertensive rat; VSMC: vascular smooth muscle cell; CDK4: cyclin-dependent kinase 4; pRb: phospho-retinoblastoma protein."

[1] Laurent S, Vanhoutte P, Cavero I, Chabrier PE, Dupuis B, Elghozi JL, Hamon G, Janiak P, Juillet Y, Kher A, Koen R, Madonna O, Maffrand JP, Pruneau D, Thuillez C . The arterial wall : a new pharmacological and therapeutic target. Fundam Clin Pharmacol. 1996; 10(3) : 243-257.
[2] Arterial aging and hypertension (to be continued) . Zhonghua Gao Xue Ya Za Zhi. 2007; 15(10) : 866-867. Chinese.
动脉老化与髙血压( 待续). 中华髙血压杂志. 2007; 15(10) : 866-867.
[3] Yang J, Lei Y, Fang SP, Cui W , Chen KJ. Study on acting mechanism of extracts from ginseng, notoginseng and Chuanxiong for delaying the aging of endothelial cells induced by angiotensin. Zhongguo Zhong Xi Yi Jie He Za Zhi. 2009; 29(6):524-528. Chinese with abstract in English.
杨静, 雷燕, 方素萍, 崔巍, 陈可冀 . 人参三七川芎提取物延缓血管紧张素n诱导内皮细胞衰老机制的研究. 中国中西医结合杂志. 2009; 29(6):524-528.
[4] Ma L, Liu SJ, Deng YZ, Sun ZQ , Li YF. A method to isolate, culture and identify smooth muscle cells from rat thoracic artery. Zhongguo Xin Xue Guan Bing Yan Jiu Za Zhi. 2007 ; 5(5) : 363-365. Chinese with abstract in English.
马丽, 刘苏健, 邓勇志, 孙宗全, 栗一帆 . 大鼠胸主动脉平滑肌细胞的培养与鉴定.中国心血管病研究杂志. 2007; 5(5) : 363-365.
[5] Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, Medrano EE, Linskens M, Rubelj I, Pereira-Smith O, Peacocke M, Campisi J . A biomarker that identifies senescent human cells in culture and in aging skin in vivo . Proc Natl Acad Sci USA. 1995; 92(20) : 9363-9367.
[6] Situ ZQ, Wu JZ. Cell culture. 2nd ed. Xi'an: World Publishing Corporation. 2007: 200-201. Chinese.
司徒镇强, 吴军正 .细胞培养. 第2版.西安: 世界图书出版公司. 2007: 200-201.
[7] Varagic J, Susie D, Frohlich E . Heart, aging and hypertension. Curr Opin Cardiol. 2001 ; 16(6):336-341.
[8] Ruan YJ, Yin MZ , Wu SZ. Research on vascular aging. Shi Yong Lao Nian Yi Xue. 2006; 20(6):418-420. Chinese.
阮云军, 银孟卓, 吴赛珠 . 血管老化的研究进展.实用老年医学. 2006; 20(6):418-420.
[9] Wen JK, Han M. Supplementing qi and activating blood circulation in gene regulation of vascular smooth muscle cells. J Chin Integr Med. 2003; 1(1):12-14. Chinese with abstract in English.
温进坤, 韩梅 . 益气活血法与血管平滑肌细胞基因表达调节.中西医结合学报. 2003; 1(1):12-14.
[10] JinJS, Zhao CH, Chen XC, Zhu YG, Zeng YQ, Li YK, Peng XS , Shi GB. Antiaging effect of ginsenoside Rgl is related with expressions of pi 6, cyclin D1 and CDK4. Zhongguo Lin Chuang Yao Li Xue Yu Zhi Liao Xue. 2004; 9(1):29-34. Chinese with abstract in English.
金建生, 赵朝晖, 陈晓春, 朱元贵, 曾育琦, 李永坤, 彭小松, 师广斌 . 人参阜苷Rgl抗衰老作用可能与改变pl6, cyclinD,CDK4的表达有关.中国临床药理学与治疗学. 2004; 9(1):29-34.
[11] Chen XC, Zhu YG, Wang XZ, Zhu LA, Huang C . Protective effect of ginsenoside Rgl on dopamine- induced apoptosis in PCI2 cells. Acta Pharmacol Sin. 2001; 22(8) : 673-678.
[12] Xue LX, Li J, Zhang ZY, Tong TJ. Cell senescence and transcriptional regulation of pl6 INK4a. Yi Xue Fen Zi Sheng Wu Xue Za Zhi. 2006; 3(1):29-31. Chinese with abstract in English.
薛丽香, 李琚, 张宗玉, 童坦君 . 细胞衰老与pl6 INK4a 的转录调控.医学分子生物学杂志 . 2006; 3(1):29-31.
[13] Hu ZW, Zhou YP , Shen ZY. Research progress of the relationship between pi6 and cell senescence. Guo Wai Yi Xue Yi Chuan Xue Fen Ce. 2004; 27(4):200-202. Chinese.
胡作为, 周燕萍, 沈自尹 . Pl6基因与细胞衰老关系的研究进展.国外医学遗传学分册. 2004; 27(4):200-202.
[14] Ou Q, Wang YL, Wei XD , Zhang YC. Effect of Fructus corni polysaccharides on the activity of p-galactosidase and the expression of pi 6. Zhongguo Lao Nian Xue Za Zhi. 2007 ; 4(27):325-327. Chinese with abstract in English.
欧芹, 王雅丽, 魏晓东, 张玉成 . 山茱萸多糖对HDF 细胞卩-半乳糖苷酶及Pl6表达的影响.中国老年学杂志. 2007; 4(27):325-327.
[15] Thomas NS, Pizzey AR, Tiwari S, Williams CD , Yang J. pi30,pl07, and pRb are differentially regulated in proliferating cells and during cell cycle arrest by a-inter- feron.J Biol Chem. 1998; 273(37):23659-23667.
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