Search JIM Advanced Search

Journal of Chinese Integrative Medicine ›› 2012, Vol. 10 ›› Issue (11): 1286-1292.doi: 10.3736/jcim20121113

• Original Experimental Research • Previous Articles     Next Articles

Effects of Chinese herbal medicine Xiaopi Pill in preventing rats from dimethylnitrosamine-induced liver fibrosis

Xiao Zhang1, Bing-bing Ning1, Shuang Ren1, Li-jun Zhang1, Wen-meng Zhang2, Jia-mei Chen1, Gao-feng Chen1, Hua Zhang1, Yong-ping Mu1, Ping Liu1,3()   

  1. 1. Institute of Liver Diseases, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
    2. School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, Liaoning Province, China
    3. E-Institute of Shanghai Municipal Education Commission, Shanghai Municipal Education Commission, Shanghai 201203, China
  • Received:2012-06-01 Accepted:2012-06-28 Online:2012-11-20 Published:2018-11-15

Objective: To explore the intervention effects of Xiaopi Pill (XPW), a compound traditional Chinese herbal medicine, on the development progress of dimethylnitrosamine (DMN)-induced liver fibrosis in rats.

Methods: Liver fibrosis model was established by intraperitoneal injection of 0.5% DMN 2 mL/kg thrice a week for 4 weeks. Rats were divided into control group given saline and treatment group given XPW during the 3rd week of DMN injections. Rats were sacrificed at the end of the experiment, and then liver histological changes, liver function and mRNA expression of the liver fibrosis-associated markers were observed.

Results: (1) At the end of the 2nd and 4th weeks of DMN injection, serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) increased significantly in rats (P<0.01 orP<0.05); content of total bilirubin (TBil) increased significantly compared with the normal group until the end of the 4th week (P<0.05); compared with the model group after 4 weeks of DMN injection, the serum levels of ALT, AST, ALP and TBil were decreased remarkably in the XPW-treated group (P<0.01 or P<0.05). (2) The hepatic inflammation and collagen deposition in hepatic tissues increased by different degrees in experimental rats. Parts of pathological changes in the rat liver were found at the end of the 4th week, including a complete round structure of false flocculus round, meantime, the hydroxyproline content of hepatic tissue was increased significantly at the end of the 2nd and 4th weeks (P<0.05). Compared with the 4-week model group, the hepatic inflammation, collagen deposition and hydroxyproline content in hepatic tissues were alleviated dramatically (P<0.05). (3) Compared with the normal and 2nd week groups, protein expression of alpha-smooth muscle actin (α-SMA) was gradually increased, and that of the 4th week group were aggrandized significantly (P<0.01). Compared with the normal group, the mRNA expression of α-SMA, transforming growth factor-β1 (TGF-β1), tissue inhibitor of melalloproteinase-1 (TIMP-1), and heme oxygenase-1 (HO-1) was gradually increased. Further changes in above-mentioned abnormalities were found in the model rats at the end of the 4th week (P<0.01); while compared to the 4th week group, protein and mRNA levels of α-SMA and mRNA levels of TGF-β1, TIMP-1, and HO-1 were decreased significantly in the XPW group (P<0.01 or P<0.05).

Conclusion: Progressive DMN-induced liver fibrosis in rats can be suppressed by XPW; the mechanism may be associated with inhibition of the activated hepatic stellate cells.

Key words: drugs,Chinese herbal, liver cirrhosis, experimental, dimethylnitrosamine, liver function tests, hydroxyproline, rats

Table 1

Primer sequences of real-time polymerase chain reaction"

Primer name Sequence Note

Figure 1

Pathological changes of liver tissues (a): Photographs of hematoxylin-eosin staining (Light microscopy, ×200); (b): Photographs of Sirius red staining (Light microscopy, ×100). A: Normal group; B: 2-week model group; C: 4-week model group; D: Xiaopi Pill group."

Table 2

Liver function of rats in different groups (Mean±standard deviation)"

Group n ALT (IU/L) AST (IU/L) ALP (IU/L) TBil (mmol/L)
Normal 10 42.00±5.03**△△ 112.70±15.41**△△ 136.10±23.56*△△ 7.45±1.37
Two-week model 6 101.17±13.35 160.50±26.97 248.50±49.91 8.25±3.34
Four-week model 13 120.38±31.58 156.15±38.8 377.15±127.31**△△ 12.56±7.80
Xiaopi Pill 13 80.54±15.65 121.85±16.66△△ 248.38±66.81△△ 6.90±2.11

Table 3

Hydroxyproline content of liver tissues in different groups (Mean±standard deviation)"

Group n Hydroxyproline content (μg/g)
Normal 10 167.51±25.87**△△
Two-week model 6 217.76±36.79
Four-week model 13 390.74±51.34**
Xiaopi Pill 13 262.57±49.22

Table 4

Degree of collagen fiber distribution of liver fibrosis of rats in each group"

Group n Distribution of staging of liver fibrosis (n)
0 Ridit value
Normal 10 10 0 0 0 0 0.05△△
Two-week model 6 0 5 1 0 0 0.33△△
Four-week model 13 0 0 10 3 0 0.73
Xiaopi Pill 13 0 10 3 0 0 0.35△△

Figure 2

Protein expression of α-SMA in liver tissues (a) Expression of α-SMA protein observed by immunohistochemical method (Light microscopy, ×200). (b) Expression of α-SMA protein observed by Western blotting. (c) The gray-level score of Western blotting results. △△P<0.01, vs four-week model group. A: Normal group; B: 2-week model group; C: 4-week model group; D: Xiaopi Pill group; α-SMA: alpha-smooth muscle actin; GAPDH: glyceraldehyde-3-phosphate dehydrogenase."

Figure 3

mRNA expressions of α-SMA, TGF-β1, TIMP-1 and HO-1 in liver tissues △P<0.05, △△P<0.01, vs four-week model group. A: Normal group; B: 2-week model group; C: 4-week model group; D: Xiaopi Pill group. α-SMA: alpha-smooth muscle actin; TGF-β1: transforming growth factor-β1; TIMP-1: tissue inhibitor of metalloproteinase-1; HO-1: heme oxygenase-1."

[1] Ye F, Xue BY, Zhou M, Zhou ZY. Damp-heat toxin accumulation and hepatofibrosis. Nanjing Zhong Yi Yao Da Xue Xue Bao. 2005; 21(6):346-349 Chinese.
doi: 10.3969/j.issn.1000-5005.2005.06.002
叶放, 薛博瑜, 周珉, 周仲瑛 . 论湿热瘀毒与肝纤维化.南京中医药大学学报. 2005; 21(6):346-349.
doi: 10.3969/j.issn.1000-5005.2005.06.002
[2] Ala-Kokko L, Pihlajaniemi T, Myers JC, Kivirikko KI, Savolainen ER . Gene expression of type I , IE and IY collagens in hepatic fibrosis induced by dimethylnitrosamine in the rat. Biochem J. 1987; 244(1):75-79.
doi: 10.1042/bj2440075
[3] Jamall IS, Finelli VN, Que Hee SS . A simple method to determine nanogram levels of 4-hydroxyproline in biological tissues. Anal Biochem. 1981; 112(1) : 70-75.
doi: 10.1016/0003-2697(81)90261-X pmid: 7258630
[4] Nakatani K, Seki S, Kawada N, Kitada T, Yamada T, Sakaguchi H, Kadoya H, Ikeda K, Kaneda K . Expression of SPARC by activated hepatic stellate cells and its correlation with the stages of fibrogenesis in human chronic hepatitis. Virchows Arch. 2002; 441(5):466-474.
doi: 10.1007/s00428-002-0631-z
[5] Kristensen DB, Kawada N, Imamura K, Miyamoto Y, Tateno C, Seki S, Kuroki T, Yoshizato K . Proteome analysis of rat hepatic stellate cells. Hepatology. 2000; 32(2) : 268-277.
doi: 10.1053/jhep.2000.9322 pmid: 10915733
[6] Uyama N, Shimahara Y, Okuyama H, Kawada N, Kamo S, Ikeda K, Yamaoka Y . Carbenoxolone inhibits DNA synjournal and collagen gene expression in rat hepatic stellate cells in culture.[J] Hepatol. 2003; 39(5):749-755.
doi: 10.1016/S0168-8278(03)00375-1
[7] Jézéquel AM, Mancini R, Rinaldesi ML, Macarri G, Venturini C, Orlandi F . A morphological study of the early stages of hepatic fibrosis induced by low doses of dimethylnitrosamine in the rat.[J] Hepatol. 1987 ; 5(2):174-181.
doi: 10.1016/S0168-8278(87)80570-6
[8] Zhang B, Wang LT. Progress in experimental study on inhibitory effect of traditional Chinese medicine on liver fibrosis. J Chin Integr Med. 2006; 4(2): 206-210. Chinese.
doi: 10.3969/j.issn.1003-5028.2004.02.069
张斌, 王灵台 . 中医药抗肝纤维化的实验研究进展.中西医结合学报. 2006; 4(2):206-210.
doi: 10.3969/j.issn.1003-5028.2004.02.069
[9] Chinese Association of Integrative Medicine. Guideline for the diagnosis and treatment of liver fibrosis with integrative medicine. J Chin Integr Med. 2006; 4(6):551-555. Chinese.
doi: 10.7661/CJIM.2006.11.1052
中国中西医结合学会肝病专业委员会. 肝纤维化中西医结合诊疗指南.中西医结合学报. 2006; 4(6):551-555.
doi: 10.7661/CJIM.2006.11.1052
[10] Okada A, Gamier JM, Vicaire S, Basset P . Cloning of the cDNA encoding rat tissue inhibitor of metalloproteinase 1 (TIMP-1), amino acid comparison with other TIMPs, and gene expression in rat tissues. Gene. 1994; 147(2):301-302.
doi: 10.1016/0378-1119(94)90088-4
[11] Yoshiji H, Kuriyama S, Yoshii J, Ikenaka Y, Noguchi R, Nakatani T, Tsujinoue H, Yanase K, Namisaki T, Imazu H, Fukui H . Tissue inhibitor of metalloproteinases- 1 attenuates spontaneous liver fibrosis resolution in the transgenic mouse. Hepatology. 2002 ; 36(4) : 850-860.
doi: 10.1053/jhep.2002.35625
[12] Wang XH, Cao Q, Li J, Huang Y. Thefunction regulation and development of heme oxygenase-1 in the liver disease. Zhongguo Yao Li Xue Tong Bao. 2006 ; 22(5) : 525-528. Chinese with abstract inEnglish.
doi: 10.3321/j.issn:1001-1978.2006.05.004
王晓红, 曹琦, 李俊, 黄艳 . 血红素氧化酶功能和调节及其在肝病发生中的研究进展.中国药理学通报. 2006; 22(5) : 525-528.
doi: 10.3321/j.issn:1001-1978.2006.05.004
[13] Huang XF, Chen BC, Hu ZY, Lin LN, Luo AL. Protective role of heme oxygenase-1 on the liver ischemia/ reperfusion injury. Gan Dan Yi Wai Ke Za Zhi. 2006; 18(2):95-96. Chinese with abstract in English.
doi: 10.3969/j.issn.1007-1954.2006.02.011
黄秀芳, 陈必成, 胡正杨, 林丽娜, 罗爱林 . 血红素加氧酶-1诱导对鼠肝缺血再灌注损伤的保护作用.肝胆胰外科杂志. 2006; 18(2):95-96.
doi: 10.3969/j.issn.1007-1954.2006.02.011
[14] Li YM, Zhang P, Zhang Y, Zhang QS, Wu LL, Wu YJ. Alteration and role of heme oxygenase in liver during sepsis. Chengde Yi Xue Yuan Yuan Bao. 2001 ; 18(3):202-205. Chinese with abstract in English.
李玉明, 张鹏, 张游, 张秋实, 吴立玲, 伍贻经 . 脓毒血症大鼠肝脏血红素加氧酶的变化及作用.承德医学院院报. 2001; 18(3):202-205.
[1] Bing-rong Li, Shi-yun Shao, Long Yuan, Ru Jia, Jian Sun, Qing Ji, Hua Sui, Li-hong Zhou, Yi Zhang, Hui Liu, Qi Li, Yan Wang, Bi-meng Zhang. Effects of mild moxibustion on intestinal microbiome and NLRP3 inflammasome in rats with 5-fluorouracil-induced intestinal mucositis. Journal of Integrative Medicine, 2021, 19(2): 144-157.
[2] Sitthichai Iamsaard, Supatcharee Arun, Jaturon Burawat, Supataechasit Yannasithinon, Saranya Tongpan, Sudtida Bunsueb, Natthapol Lapyuneyong, Pannawat Choowong-in, Nareelak Tangsrisakda, Chadaporn Chaimontri, Wannisa Sukhorum. Evaluation of antioxidant capacity and reproductive toxicity of aqueous extract of Thai Mucuna pruriens seeds. Journal of Integrative Medicine, 2020, 18(3): 265-273.
[3] Bérénice Colleville, Nicolas Perzo, Guillaume Avinée, Anaïs Dumesnil, Frederic Ziegler, Paul Billoir, Hélène Eltchaninoff, Vincent Richard, Eric Durand. Impact of high-fat diet and vitamin D3 supplementation on aortic stenosis establishment in waved-2 epidermal growth factor receptor mutant mice. Journal of Integrative Medicine, 2019, 17(2): 107-114.
[4] Morufu Eyitayo Balogun, Elizabeth Enohnyaket Besong, Jacinta Nkechi Obimma, Ogochukwu Sophia Mbamalu, Fankou Serges Athanase Djobissie. Protective roles of Vigna subterranea (Bambara nut) in rats with aspirin-induced gastric mucosal injury. Journal of Integrative Medicine, 2018, 16(5): 342-349.
[5] Lucky Legbosi Nwidu, Raphael Ellis Teme. Hot aqueous leaf extract of Lasianthera africana (Icacinaceae) attenuates rifampicin-isoniazid-induced hepatotoxicity. Journal of Integrative Medicine, 2018, 16(4): 263-272.
[6] Kylie Connolly, Douglas Jackson, Candice Pullen, Andrew Fenning. Alpha-adrenoceptor antagonism by Crassostrea gigas oyster extract inhibits noradrenaline-induced vascular contraction in Wistar rats. Journal of Integrative Medicine, 2015, 13(3): 194-200.
[7] Udhaya Lavinya Baskaran, Sherry Joseph Martin, Rasool Mahaboobkhan, Sabina Evan Prince. Protective role of Triphala, an Indian traditional herbal formulation, against the nephrotoxic effects of bromobenzene in Wistar albino rats. Journal of Integrative Medicine, 2015, 13(2): 115-121.
[8] Hussein O. B. Oloyede, Matthew C. Adaja, Taofeek O. Ajiboye, Musa O. Salawu. Anti-ulcerogenic activity of aqueous extract of Carica papaya seed on indomethacin-induced peptic ulcer in male albino rats. Journal of Integrative Medicine, 2015, 13(2): 105-114.
[9] Anirudha A. Lande, Shirishkumar D. Ambavade, Uma S. Swami, Prafulla P. Adkara Prashant D. Ambavade, Arun B. Waghamare. Saponins isolated from roots of Chlorophytum borivilianum reduce acute and chronic inflammation and histone deacetylase. Journal of Integrative Medicine, 2015, 13(1): 25-33.
[10] Elizabeth Abidemi Balogun, Sylvia Orume Malomo, Joseph Oluwatope Adebayo, Ahmed Adebayo Ishola, Ayodele Olufemi Soladoye, Lawrence Aderemi Olatunji, Olatunji Matthew Kolawole, Stephen Olubunmi Oguntoye, Abiola Samuel Babatunde, Oluwole Busayo Akinola . In vivo antimalarial activity and toxicological effects of methanolic extract of Cocos nucifera (Dwarf red variety) husk fibre. Journal of Integrative Medicine, 2014, 12(6): 504-511.
[11] Urmila Aswar, Mayuri Gurav, Ganesh More, Khaled Rashed, Manoj Aswar. Effect of aqueous extract of Solanum xanthocarpum Schrad. & Wendl. on postmenopausal syndrome in ovariectomized rats. Journal of Integrative Medicine, 2014, 12(5): 439-446.
[12] Chang-qing Zhao, Yang Zhou, Jian Ping, Lie-ming Xu. Traditional Chinese medicine for treatment of liver diseases: Progress, challenges and opportunities. Journal of Integrative Medicine, 2014, 12(5): 401-408.
[13] Xin-fang Zhang, Ji Zhu, Wen-ye Geng, Shu-jun Zhao, Chuan-wei Jiang, Sheng-rong Cai, Miao Cheng, Chuan-yun Zhou, Zi-bing Liu. Electroacupuncture at Feishu (BL13) and Zusanli (ST36) down-regulates the expression of orexins and their receptors in rats with chronic obstructive pulmonary disease. Journal of Integrative Medicine, 2014, 12(5): 417-424.
[14] Benjamin Perry, Junzeng Zhang, Tarek Saleh, Yanwen Wang. Liuwei Dihuang, a traditional Chinese herbal formula, suppresses chronic inflammation and oxidative stress in obese rats. Journal of Integrative Medicine, 2014, 12(5): 447-454.
[15] Upendarrao Golla, Praveen Kumar Gajam, Solomon Sunder Bhimathati. Evaluation of diuretic and laxative activity of hydro-alcoholic extract of Desmostachya bipinnata (L.) Stapf in rats. Journal of Integrative Medicine, 2014, 12(4): 372-378.
Full text



[1] Jin-zhou Tian, Jing Shi, Xin-qing Zhang, Qi Bi, Xin Ma, Zhi-liang Wang, Xiao-bin Li, Shu-li Shen, Lin Li, Zhen-yun Wu, Li-yan Fang, Xiao-dong Zhao, Ying-chun Miao, Peng-wen Wang, Ying Ren, Jun-xiang Yin, Yong-yan Wang, Beijing United Study Group on MCI of the Capital Foundation of Medical Developments. An explanation on "guiding principles of clinical research on mild cognitive impairment (protocol)". Journal of Chinese Integrative Medicine, 2008, 6(1): 15-21
[2] Yi-ting He, Qing-lin Zha, Jian-ping Yu, Yong Tan, Cheng Lu, Ai-ping Lv. Principal factor analysis of symptoms of rheumatoid arthritis and their correlations with efficacy of traditional Chinese medicine and Western medicine. Journal of Chinese Integrative Medicine, 2008, 6(1): 32-36
[3] Jun Cai, Hua Wang, Sheng Zhou, Bin Wu, Hua-rong Song, Zheng-rong Xuan. Effect of Sijunzi Decoction and enteral nutrition on T-cell subsets and nutritional status in patients with gastric cancer after operation: A randomized controlled trial. Journal of Chinese Integrative Medicine, 2008, 6(1): 37-40
[4] Wei Zhang, Xiang-feng Lu, Xiao-mei Zhang, Jian-jun Wu, Liang-duo Jiang. A rat model of pulmonary fibrosis induced by infusing bleomycin quickly through tracheal intubation. Journal of Chinese Integrative Medicine, 2008, 6(1): 60-67
[5] A-gao Zhou, Yong Zhang, Gang Kui, De-Yun Kong, Hai-liang Ge, Qiu-hua Ren, Jia-rong Dong, Sheng Hong, Xu-ming Mao, Yin Wang, Hui-zheng Zhang, Shu-jun Wang. Influence of traditional Chinese compound recipes with different efficacy on body weight, tumor weight and immune function in H22 cancer-bearing mice. Journal of Chinese Integrative Medicine, 2008, 6(1): 77-82
[6] Guo-hong Yuan, Xiao-jing Pang, He-chao Ma. Synergic effects of Danggui Buxue Decoction in reducing toxicity of cytoxan in tumor-bearing mice. Journal of Chinese Integrative Medicine, 2008, 6(1): 83-88
[7] Li Zhou, Hong-xing Zhang, Ling-guang Liu, Wen-jun Wan. Effect of electro-acupuncture at Fenglong (GV 16) on nitric oxide and endothelin in rats with hyperlipidemia. Journal of Chinese Integrative Medicine, 2008, 6(1): 89-92
[8] Jin-zhou Tian, Jing Shi, Xin-qing Zhang, Qi Bi, Xin Ma, Zhi-liang Wang, Xiao-bin Li, Shu-li Shen, Lin Li, Zhen-yun Wu, Li-yan Fang, Xiao-dong Zhao, Ying-chun Miao, Peng-wen Wang, Ying Ren, Jun-xiang Yin, Yong-yan Wang, Beijing United Study Group on MCI of the Capital Foundation of Medical Developments. Guiding principles of clinical research on mild cognitive impairment (protocol). Journal of Chinese Integrative Medicine, 2008, 6(1): 9-14
[9] Xin-jun Wang, Ling-ling Wang . A mechanism of endogenous opioid peptides for rapid onset of acupuncture effect in treatment of depression. Journal of Chinese Integrative Medicine, 2010, 8(11): 1014-1017
[10] Bo Wang , Wei Yan , Li-hui Hou, Xiao-ke Wu. Disorder of Tiangui (kidney essence) and reproductive dysfunction in patients with polycystic ovary syndrome. Journal of Chinese Integrative Medicine, 2010, 8(11): 1018-1022