Evidence-Based Complementary and Alternative Medicine

Research Article

Synergistic Cytotoxic Effects of Ganoderma lucidum and Bacillus Calmette Guérin on Premalignant Urothelial HUC-PC Cells and Its Regulation on Proinflammatory Cytokine Secretion

Table 1

Yuen et al.
Treatment scheduleCytokine level (Mean ± SD); 𝑛 = 3
IL-2 (pg/mL)IL-6 (pg/mL)IL-8 (pg/mL)MCP-1 (pg/mL)
Treatment 1 (24 hours)     
BCG alone0 CFU†N.D. 7 6 ± 2 1 7 4 4 ± 1 9 3 1 8 1 2 ± 1 2 4
  1 . 2 × 1 0 7  CFUN.D. 3 8 0 ± 2 5 *** 1 6 2 9 ± 4 1 6 *** 1 8 5 0 ± 2 5 8
  2 . 4 × 1 0 7  CFUN.D. 4 9 2 ± 2 9 *** 1 5 8 0 ± 5 0 3 *** 1 6 6 1 ± 2 7 8
  4 . 8 × 1 0 7  CFUN.D. 4 9 8 ± 3 5 *** 1 6 5 1 ± 7 0 8 *** 1 4 9 0 ± 1 7 0 ***
    ( 𝐹 = 1 0 9 3 ) ( 𝐹 = 2 0 . 8 2 ) ( 𝐹 = 5 . 0 4 4 )
Treatment 2 (24 hours)     
GLe alone0 μg/mL† 6 ± 1 1 5 7 ± 2 4 6 4 5 ± 1 0 2 1 7 6 5 ± 1 1 5
 40 μg/mL 7 ± 1 3 3 3 1 ± 3 7 *** 1 5 8 1 ± 3 6 7 *** 1 2 5 4 ± 1 0 1 ***
 80 μg/mL 1 0 ± 1 9 8 2 2 ± 3 5 *** 2 4 3 0 ± 3 9 7 *** 7 4 0 ± 7 1 ***
 100 μg/mL 2 2 ± 2 9 1 3 4 3 ± 1 3 0 *** 3 0 9 1 ± 6 7 2 *** 6 7 1 ± 6 2 ***
    ( 𝐹 = 1 . 2 3 8 ) ( 𝐹 = 5 4 1 . 6 ) ( 𝐹 = 1 0 5 . 2 ) ( 𝐹 = 2 8 0 . 6 )
Treatment 3 (24 hours)     
BCG + GLe  
0  𝜇 g/mL†		N.D. 4 1 3 ± 2 0 2 1 8 3 5 ± 8 0 8 1 7 8 9 ± 3 0 8
1 . 2 × 1 0 7  CFU40  𝜇 g/mLN.D. 7 5 2 ± 3 9 * 1 7 3 3 ± 4 6 9 5 2 0 ± 1 3 7 ***
  
80  𝜇 g/mL		N.D. 7 5 4 ± 8 3 *** 1 5 0 4 ± 3 2 8 8 1 8 ± 1 6 9 ***
  
100  𝜇 g/mL		N.D. 8 9 1 ± 4 0 *** 1 4 3 6 ± 3 3 8 5 4 2 ± 1 0 1 ***
      ( 𝐹 = 1 1 . 1 2 ) ( 𝐹 = 1 . 6 2 8 ) ( 𝐹 = 4 2 . 8 9 )
Treatment 4 (48 hours)      
24-hr GLe → 24-hr BCG 0  𝜇 g/mL†N.D. 6 4 0 ± 4 4 2 2 5 6 ± 8 3 0 3 4 7 1 ± 3 3 8
1.2 × 107 CFU40  𝜇 g/mLN.D. 2 7 0 ± 2 7 *** 8 7 4 ± 6 8 *** 9 6 2 ± 1 0 7 ***
 80  𝜇 g/mLN.D. 1 6 4 ± 3 1 *** 5 2 4 ± 2 4 3 *** 8 9 ± 1 0 3 ***
 100  𝜇 g/mLN.D. 1 6 1 ± 2 0 *** 6 3 4 ± 2 0 6 *** N.D.***
      ( 𝐹 = 8 2 7 . 2 )( 𝐹 = 4 3 . 4 5 ) ( 𝐹 = 2 6 9 . 3 )
The secretion of cytokines detectable in the conditioned media collected from the cells treated with different treatment schedules (N.D.: non-detectable; * 𝑃 < 0.05; *** 𝑃 < 0.001). Statistical significances of parameters in each treatment schedule group were compared with corresponding control†. Controls for treatment 1 and 2 were not identical, because the 0 CFU BCG in treatment 1 contains 33% (v/v) BCG diluents, whereas the 0 𝜇 g/mL GLe in treatment 2 contains 0.1% (v/v) ethanol. Additionally, the diluting effect of BCG diluents in the complete media was suspected to be the cause of non-detectable level of IL-2 in treatment 1, as compared with the trace level produced in the GLe control in treatment 2. However, the secretion of other cytokines (IL-6, IL-8 and MCP-1 were not in trace amounts) was seemed to be not being affected by such dilution. 𝐹 value of each treatment group was determined by one-way ANOVA, and Dunnett’s post test was followed to determine 𝑃 values.