title: The combination effects of Shen-Ling-Bai-Zhu on promoting apoptosis of transplanted H22 hepatocellular carcinoma in mice receiving chemotherapy.
abstract: BACKGROUND
  Shen-Ling-Bai-Zhu Powder (SLBZP) is a classic traditional Chinese medical formula that has been used for several decades in the treatment of patients with gastrointestinal malignancies. Whether SLBZP is best employed as single agent or adjunctive therapy has yet to be determined as does the mechanism whereby SLBZP exerts its anti-tumor effects.
  
  
  OBJECTIVE
  To investigate the effects of SLBZP alone and in combination with Cytoxan (CTX) on tumor growth, malignant cell apoptosis and Akt/Nuclear Factor kappa B (NF-КB) signaling in a murine model of hepatocellular carcinoma (HCC) receiving chemotherapy.
  
  
  METHODS
  Sixty-four adult mice developed HCC following subcutaneous inoculation with H22 hepatocellular carcinoma cells. Seven days later, all received chemotherapy with CTX (200mg/kg) once. Mice were then randomized into eight study groups (N=8/group). Three groups were treated with different concentrations of SLBZP alone (6.00, 3.00, 1.5g/kg), three with SLBZP (6.00, 3.00, 1.5g/kg) plus CTX (20mg/kg), one with CTX (20mg/kg) alone (positive control), and one with physiologic saline (untreated, negative control). All groups were treated for 14 days. Tumor size, histology and serum or tissue levels and/or mRNA expression of PDGF-BB, VEGF, Ang-1, Ang-2, NF-КB, B-cell lymphoma-2 (Bcl-2); B-cell lymphoma-extra large (Bcl-xL); X-linked inhibitor of apoptosis (XIAP), Survivin, Caspase-3, Caspase-9, Caspase-7, Akt and phosphorylated Akt expression were documented at the end of treatment.
  
  
  RESULTS
  Compared to untreated negative controls, tumor sizes were decreased in the CTX alone, SLBZP (M)+CTX and SLBZP (H)+CTX groups (-52%,-53% and -58% respectively). Tumor cell density was decreased in all treated groups but most apparent in the SLBZP (H)+CTX group. Electron microscopic evidence of apoptosis was also most apparent in this group. Serum and/or tissue levels and expression of PDGF-BB, VEGF, Ang-1, Ang-2, their downstream signaling proteins and anti-apoptotic markers were lowest and pro-apoptotic markers highest in SLBZP (H)+CTX treated mice.
  
  
  CONCLUSIONS
  In this chemotherapy-induced animal model of HCC, SLBZP was most efficacious as adjunctive therapy and appears to act by inhibiting tumor growth promoters and anti-apoptotic proteins while enhancing pro-apoptotic proteins.
authors.affiliation: Department of Traditional Chinese Medicine, Medical College, Xiamen University, Xiamen 361102, People's Republic of China; Cancer Research Center of Xiamen University, Xiamen 361102, People's Republic of China. Electronic address: xishengyan@xmu.edu.cn.
authors.forename: Shengyan
authors.initials: S
authors.lastname: Xi
authors.affiliation: Department of Traditional Chinese Medicine, Medical College, Xiamen University, Xiamen 361102, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Peng
authors.affiliation: Department of Internal Medicine, Faculty of Health Sciences, University of Manitoba, Winnipeg R3E 3P4, Manitoba, Canada.
authors.forename: Gerald Y
authors.initials: GY
authors.lastname: Minuk
authors.affiliation: Department of Traditional Chinese Medicine, Medical College, Xiamen University, Xiamen 361102, People's Republic of China.
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Shi
authors.affiliation: Department of Traditional Chinese Medicine, Medical College, Xiamen University, Xiamen 361102, People's Republic of China.
authors.forename: Biqian
authors.initials: B
authors.lastname: Fu
authors.affiliation: College of Pharmacy, University of Manitoba, Winnipeg R3E 0T5, Manitoba, Canada.
authors.forename: Jiaqi
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Internal Medicine, Faculty of Health Sciences, University of Manitoba, Winnipeg R3E 3P4, Manitoba, Canada.
authors.forename: Qian
authors.initials: Q
authors.lastname: Li
authors.affiliation: College of Pharmacy, University of Manitoba, Winnipeg R3E 0T5, Manitoba, Canada.
authors.forename: Yuewen
authors.initials: Y
authors.lastname: Gong
authors.affiliation: Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing 100700, People's Republic of China.
authors.forename: Lifeng
authors.initials: L
authors.lastname: Yue
authors.affiliation: Department of Traditional Chinese Medicine, Medical College, Xiamen University, Xiamen 361102, People's Republic of China.
authors.forename: Lili
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Traditional Chinese Medicine, Medical College, Xiamen University, Xiamen 361102, People's Republic of China.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Guo
authors.affiliation: College of Pharmacy, University of Manitoba, Winnipeg R3E 0T5, Manitoba, Canada.
authors.forename: Yang
authors.initials: Y
authors.lastname: Peng
authors.affiliation: Department of Traditional Chinese Medicine, Medical College, Xiamen University, Xiamen 361102, People's Republic of China.
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Wang
mesh_terms: D042501:Angiogenic Proteins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D008114:Liver Neoplasms, Experimental / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D047368:Tumor Burden / Q000187:drug effects
chemical_list: D042501:Angiogenic Proteins; D000972:Antineoplastic Agents, Phytogenic; D051017:Apoptosis Regulatory Proteins; D014408:Biomarkers, Tumor; D004365:Drugs, Chinese Herbal; D012333:RNA, Messenger; C532982:shen ling bai zhu; D002945:Cisplatin
keywords: Akt; Apoptosis; Bcl-2, Bcl-XL; Butenolide A (Atractylenolide I) (PubChem CID: 5321018 ); Butenolide B (Atractylenolide II ) (PubChem CID: 14448070); Caspase-3; Caspase-9; Chemotherapy; Diosgenin (PubChem CID: 99474); Ginsenoside Rb1 (PubChem CID: 9898279); Ginsenoside Rg1 (PubChem CID: 441923); H(22) hepatocellular carcinoma; Monoammonium glycyrrhetate (PubChem CID: 201991); Nuclear factor kappa B; Pachymic acid (PubChem CID: 5484385); Platelet-derived growth factor; Polyporenic acid C (PubChem CID: 9805290); Shen-Ling-Bai-Zhu powder; Survivin