Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (1): 65-69.doi: 10.3736/jcim20090110

• Original Experimental Research • Previous Articles     Next Articles

Effects of ligustrazine injection on high glucose-induced type Ⅰ collagen, matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 expressions in human peritoneal mesothelial cells in vitro

 Gui-song Zhu, Jin-song He   

  1. Department of Nephrology, the Affiliated Drum Tower Hospital, Nanjing University Medical School, Nanjing, Jiangsu Province 210008, China
  • Received:2008-06-18 Accepted:2008-07-22 Online:2009-01-20 Published:2009-01-15


To investigate the effects of ligustrazine injection on type Ⅰ collagen, matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) expressions in human peritoneal mesothelial cells (HPMCs) cultured in high glucose conditions.

 HPMCs were isolated from human omenta by trypsin digestion method and subcultured. Then, the HPMCs were divided into normal control group, high glucose group and high glucose plus low-, medium- and high-dose ligustrazine (10, 20 and 40 mg/L ligustrazine respectively) groups. Semi-quantitative reverse transcription-polymerase chain reaction was used to detect the expressions of type Ⅰ collagen, MMP-1 and TIMP-1 mRNAs in HPMCs. Proteins of type Ⅰ collagen, MMP-1 and TIMP-1 in culture supernatants were measured by enzyme-linked immunosorbent assay (ELISA). Cell protein concentration was measured by trace bicinchoninic acid method to correct the ELISA assay results.

Ligustrazine injection could significantly decrease high glucose-induced type Ⅰ collagen and TIMP-1 expressions in a dose-dependent manner both in protein and gene levels (P<0.05, P<0.01). In addition, medium- and high-dose ligustrazine injection could significantly increase MMP-1 expression which was inhibited by high glucose concentrations (P<0.05).

 Ligustrazine injection does not only decrease type Ⅰ collagen synthesis, but also promote its degradation by modulating unbalanced MMP-1/TIMP-1 expression in HPMCs cultured in high glucose conditions.

Key words: Ligustrazine injection, Human peritoneal mesothelial cells, High glucose, Type I collagen, Matrix metalloproteinase-1, Tissue inhibitor of metalloproteinase-1, In vitro

Table 1

The primer sequences and PCR conditions of type Ⅰ collagen, MMP-1, TIMP-1 and β-actin"

Target gene Primer sequence Fragment size (bp) PCR condition
Type Ⅰ collagen forward 5'-TCC AAA GGA GAG AGC GGT AA-3' 693 95 ℃ 2 min, 94 ℃ 30 s, 56 ℃ 30 s, 72 ℃ 1 min, 72 ℃ 10 min, 30 cycles
reverse 5'-GAC CAG GGA GAC CAA ACT CA-3'
MMP-1 forward 5'-CAT CCA AGC CAT ATA TGG ACG TTC C-3' 611 95 ℃ 1 min, 94 ℃ 30 s, 60 ℃ 1 min, 72 ℃ 1.5 min, 72 ℃ 10 min, 32 cycles
TIMP-1 forward 5'-TGC ACC TGT GTC CCA CCC CAC CCA CAG ACG-3' 551 94 ℃ 3 min, 94 ℃ 50 s, 58 ℃ 50 s, 72 ℃ 1 min, 72 ℃ 10 min, 35 cycles
β-actin forward 5'-TCG ACA ACG GCT CCG GCA T-3' 241 95 ℃ 3 min, 94 ℃ 40 s, 56 ℃ 30 s, 72 ℃ 1 min, 72 ℃ 10 min, 28 cycles

Table 2

Effects of ligustrazine on type Ⅰ collagen, MMP-1 and TIMP-1 protein levels in culture supernatants (x±s, ng/mg protein)"

Group n Type Ⅰ collagen MMP-1 TIMP-1
Normal control 9 1.15±0.40 12.16±0.35 0.73±0.03
High glucose 9 6.63±0.08** 7.75±0.21** 6.69±0.27**
10 mg/L ligustrazine 9 5.99±0.26 7.90±0.42 4.98±0.06△△
20 mg/L ligustrazine 9 4.46±0.28△△ 8.95±0.42△△ 3.97±0.09△△
40 mg/L ligustrazine 9 3.40±0.24△△ 10.39±0.35△△ 3.24±0.03△△

Figure 1

Expressions of type Ⅰ collagen, MMP-1 and TIMP-1 mRNAs in different groups A: Normal control group; B: High glucose group; C: 10 mg/L ligustrazine group; D: 20 mg/L ligustrazine group; E: 40 mg/L ligustrazine group."

Figure 2

Semi-quantification of type Ⅰ collagen, MMP-1 and TIMP-1 mRNAs △P<0.05, △△P<0.01, vs high glucose group."

[1] Plum J, Hermann S, Fusshller A, Schoenicke G, Donner A, Rhrborn A, Grabensee B .Peritoneal sclerosis in peritoneal dialysis patients related to dialysis settings and peritoneal transport properties[J]. Kidney Int Suppl, 2001,( 78):S42-S47
[2] Yung S, Liu ZH, Lai KN, Li LS, Chan TM . Emodin ameliorates glucose-induced morphologic abnormalities and synjournal of transforming growth factor beta1 and fibronectin by human peritoneal mesothelial cells[J]. Perit Dial Int, 2001,21(Suppl 3):S41-S47
[3] Kim JJ, Li JJ, Kim KS, Kwak SJ, Jung DS, Ryu DR, Yoo TH, Choi HY, Han SH, Kim HJ, Yoon SY, Han DS, Kang SW . High glucose decreases collagenase expression and increases TIMP expression in cultured human peritoneal mesothelial cells[J]. Nephrol Dial Transplant, 2008,23(2):534-541
[4] Perfumo F, Altieri P, Degl'Innocenti ML, Ghiggeri GM, Caridi G, Trivelli A, Gusmano R .Effects of peritoneal effluents on mesothelial cells in culture: cell proliferation and extracellular matrix regulation[J]. Nephrol Dial Transplant, 1996,11(9):1803-1809
doi: 10.1093/ndt/11.9.1803
[5] Wang YD, Yan PY . Expression of matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 in ulcerative colitis[J]. World J Gastroenterol, 2006,12(37):6050-6053
doi: 10.3748/wjg.v12.i37.6050
[6] Visse R, Nagase H . Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function, and biochemistry[J]. Circ Res, 2003,92(8):827-839
doi: 10.1161/01.RES.0000070112.80711.3D
[7] Fujisaki K, Tanabe N, Suzuki N, Mitsui N, Oka H, Ito K, Maeno M . The effect of IL-1alpha on the expression of matrix metalloproteinases, plasminogen activators, and their inhibitors in osteoblastic ROS 17/2.8 cells[J]. Life Sci, 2006,78(17):1975-1982
doi: 10.1016/j.lfs.2005.08.036
[8] Huang Y, Chen SQ, Zhang G, Chen RH . Effect of tetromethylpyrazine and aminoguanidine on expression of vascular endothelial growth factor in kidney of diabetic rats[J]. J Chin Integr Med, 2004,2(1):39-41
黄焱, 陈少强, 张更, 陈瑞华 . 川芎嗪联合氨基胍对糖尿病大鼠肾脏血管内皮细胞生长因子表达的影响[J]. 中西医结合学报, 2004,2(1):39-41
[9] Lin YZ, Xu CX, Deng YL, Chen L, Huang H, Du J . Effects of tetramethylpyrazine on fibrosis of atrial tissue and atrial fibrillation in a canine model of congestive heart failure induced by ventricular tachypacing[J]. J Chin Integr Med, 2006,4(1):35-38
林亚洲, 许春萱, 邓玉莲, 陈林, 黄海, 杜建 . 川芎嗪对心室快速起搏心力衰竭实验犬心房颤动及心房纤维化的影响[J]. 中西医结合学报, 2006,4(1):35-38
[10] Hua HY, Li YY, Ge SW . Study of tetramethylpyrazine in treatment on liver fibrosis and mechanism in rats[J]. Zhong Yao Yao Li Yu Lin Chuang, 2007,23(5):60-62
华海婴, 李艳瑛, 戈士文 . 川芎嗪抗大鼠免疫损伤性肝纤维化作用及机制研究[J]. 中药药理与临床, 2007,23(5):60-62
[11] Lin YZ, Xu CX, Huang H . The mRNA expression of collagen Ⅰ, Ⅳ and TGF-β1 in canine atrium by prevention of tetramethylpyrazine during congestive heart failure[J]. Zhong Xi Yi Jie He Xin Nao Xue Guan Bing Za Zhi, 2005,3(6):512-514
林亚洲, 许春萱, 黄海 . 川芎嗪干预心力衰竭实验犬心房组织Ⅰ、Ⅳ型胶原与TGF-β1的mRNA表达研究[J]. 中西医结合心脑血管病杂志, 2005,3(6):512-514
[12] Li JW, Liu XZ, Liu XQ, Chen HR, Lin CH, Hu EX . Effect of ligustrazine on intimal proliferation and expression of tissue inhibitor of matrix metalloproteinase-1 in balloon-injured rabbit carotid artery[J]. Zhongguo Wei Qin Xi Shen Jing Wai Ke Za Zhi, 2004,9(7):314-317
李敬文, 刘相轸, 刘晓谦, 陈会荣, 林成海, 胡恩喜 . 川芎嗪对颈动脉球囊损伤后基质金属蛋白酶组织抑制物-1基因表达的影响[J]. 中国微侵袭神经外科杂志, 2004,9(7):314-317
[1] Sherman Gu, Arthur Yin Fan. Controversial conclusions from two randomized controlled trials for acupuncture's effects on polycystic ovary syndrome or in vitro fertilization support. Journal of Integrative Medicine, 2020, 18(2): 89-91.
[2] Sana Zaki, Nasreen Jahan, Mohd Kalim, Ghausia Islam. In vitro antilithiatic activity of the hydro-alcoholic extract of Cinnamomum zeylanicum Blume bark on calcium oxalate crystallization. Journal of Integrative Medicine, 2019, 17(4): 273-281.
[3] Sahar Dehdari, Homa Hajimehdipoor, Somayeh Esmaeili, Rasool Choopani, Seyed Alireza Mortazavi. Traditional and modern aspects of hemorrhoid treatment in Iran: A review. Journal of Integrative Medicine, 2018, 16(2): 90-98.
[4] Qing-qing Zang, Lu Zhang, Ning Gao, Cheng Huang. Ophiopogonin D inhibits cell proliferation, causes cell cycle arrest at G2/M, and induces apoptosis in human breast carcinoma MCF-7 cells. Journal of Integrative Medicine, 2016, 14(1): 51-59.
[5] Mukherjee Avinaba, Boujedaini Naoual, Rahman Khuda-Bukhsh Anisur. Homeopathic Thuja 30C ameliorates benzo(a)pyrene-induced DNA damage, stress and viability of perfused lung cells of mice in vitro. Journal of Chinese Integrative Medicine, 2013, 11(6): 397-404.
[6] Dao-fang Ding, Xiao-feng Li, Hao Xu, Zhen Wang, Qian-qian Liang, Chen-guang Li, Yong-jun Wang. Mechanism of resveratrol on the promotion of induced pluripotent stem cells. Journal of Chinese Integrative Medicine, 2013, 11(6): 389-396.
[7] Yong Xiao, Zeng-chun Ma, Yu-guang Wang, Hong-ling Tan, Xiang-ling Tang, Qian-de Liang, Cheng-rong Xiao, Yue Gao. Cardioprotection of Shenfu preparata on cardiac myocytes through cytochrome P450 2J3. Journal of Chinese Integrative Medicine, 2013, 11(5): 327-336.
[8] Bolleddula Jayaprakasam, Nan Yang, Ming-Chun Wen, Rong Wang, Joseph Goldfarb, Hugh Sampson, Xiu-Min Li. Constituents of the anti-asthma herbal formula ASHMITM synergistically inhibit IL-4 and IL-5 secretion by murine Th2 memory cells, and eotaxin by human lung fibroblasts in vitro. Journal of Chinese Integrative Medicine, 2013, 11(3): 195-205.
[9] Hong Wang , Jing-tian Ou , Xin Zhao , Ying Guoz , Hao-ping Mao. Vasodilator effect of oroxylin A on thoracic aorta isolated from rats. Journal of Chinese Integrative Medicine, 2012, 10(8): 880-885.
[10] Yathiraj Ashwini, Mohamed Rafiq, Gollapalle L. Viswanatha, Subbanna Rajesh, Selvam Senthilraja, Mohammed Azeemuddin, Suryakanth D. Anturlikar, Paramesh Rangesh, Pralhad S. Patki . Polyherbal formulation Bresol® protects the mast cells against compound 48/80-induced disruption and histamine release: A non-immunological mechanism of mast cell stabilization. Journal of Chinese Integrative Medicine, 2012, 10(6): 690-694.
[11] Mukherjee Avinaba, Sikdar Sourav, Bishayee Kausik, Paul Avijit, Ghosh Samrat, Boujedaini Naoual, Rahman Khuda-Bukhsh Anisur. Ethanolic extract of Thuja occidentalis blocks proliferation of A549 cells and induces apoptosis in vitro. Journal of Chinese Integrative Medicine, 2012, 10(12): 1451-1459.
[12] Arul Raj Chinthamony, Sophia Dominic, Ragavendran Paramasivam, Starlin Thangarajan, Ahalliya Rathi Muthian, Kanniappan Gopalakrishnan Velliyur. Leaf extract of Alpinia purpurata (Vieill.) K. Schum screened for its phytochemical constituents and antibacterial and anticancer activities. Journal of Chinese Integrative Medicine, 2012, 10(12): 1460-1464.
[13] Samadder Asmita, Das Jayeeta, Das Sreemanti, Biswas Raktim, Rahman Khuda-Bukhsh Anisur. Ameliorative potentials of Syzygium jambolanum extract against arsenic-induced stress in L6 cells in vitro. Journal of Chinese Integrative Medicine, 2012, 10(11): 1293-1302.
[14] Li Zhang , Tao Wu , Jun-ming Chen , Li-li Yang , Hai-yan Song , Guang Ji. Danshensu inhibits acetaldehyde-induced proliferation and activation of hepatic stellate cell-T6. Journal of Chinese Integrative Medicine, 2012, 10(10): 1155-.
[15] Li Cai-hong, Qiu Qing-hua, Wu Xing-wei, Gong Yuan-yuan, Xie Zheng-gao, Song Yi, Gu Qing. Chinese herbal medicine Lingqi Huangban Granule protects retinal pigment epithelial cells against oxidative stress-induced injury in vitro. Journal of Chinese Integrative Medicine, 2012, 10(1): 85-90.
Full text



[1] Jin-rong Fu. Establishment of multivariate diagnosis and treatment system of modern gynecology of traditional Chinese medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 22-24
[2] Hao Li, Ming-jiang Yao, Wen-ming Zhao, Jie Guan, Lin-lin Cai, Ling Cui. A randomized, controlled, double-blind trial of Huannao Yicong capsule in senile patients with mild cognitive impairment. Journal of Chinese Integrative Medicine, 2008, 6(1): 25-31
[3] Zhi-chun Jin. Problems in establishing clinical guideline for integrated traditional Chinese and Western medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 5-8
[4] SUN Ming-yu, ZUO Jian, DUAN Ji-feng, HAN Jun, FAN Shi-ming, ZHANG Wei, ZHU Li-fang, YAO Ming-hui. Antitumor activities of kushen flavonoids in vivo and in vitro. Journal of Chinese Integrative Medicine, 2008, 6(1): 51-59
[5] Min Cheng, Qiong Feng, Shu-wen Qian, Hui Gao, Cui-qing Zhu. Preliminary assay of p-amyloid binding elements in heart-beneficial recipe. Journal of Chinese Integrative Medicine, 2008, 6(1): 68-72
[6] Ning-qun Wang, Liang-duo Jiang, Zong-xing Li. Research progress in asthma-related quality of life. Journal of Chinese Integrative Medicine, 2008, 6(1): 93-97
[7] Jing-yuan Mao, Chang-xiao Liu, Heng-he Wang, Guang-li Wei , Zhen-peng Zhang, Jie Xing, Wang Xian liang , Ying-fei Bi . Effects of Shenmai Injection on serum concentration and pharmacokinetics of digoxin in dogs with heart failure. Journal of Chinese Integrative Medicine, 2010, 8(11): 1070-1074
[8] Zhi-mei Wang, Bin Zhang. A study on translation of ellipses in Huangdi Neijing from perspective of hermeneutic theory. Journal of Chinese Integrative Medicine, 2010, 8(11): 1097-1100
[9] Gui Yu, Jie Wang. Thinking on building the network cardiovasology of Chinese medicine. Journal of Chinese Integrative Medicine, 2012, 10(11): 1206-1210
[10] Pedro Saganha João, Doenitz Christoph, Greten Tobias, Efferth Thomas, J. Greten Henry. Qigong therapy for physiotherapists suffering from burnout: a preliminary study. Journal of Chinese Integrative Medicine, 2012, 10(11): 1233-1239