Immunotoxicity induced in mice by subacute exposure to berberine

Figures & data

Figure 1. Experimental plan of doses and treatment schedules.

Table 1. Effects of subacute exposure to berberine (BBR, IP daily, 14 days) on blood cellularity.

Parameter	Vehicle	BBR 5 mg/kg^a	BBR 10 mg/kg^a	Cyclophosphamide^a
WBC (cell/µl)	9190 ± 388	8240 ± 454 (−10.3)###	6700 ± 498 (−27.0)**#	4850 ± 132 (−47.2)***
Neutrophil (cell/µl)	1976 ± 156	1505 ± 153 (−23.8)##	1349 ± 123 (−31.7)*##	2320 ± 140 (+17.4)
Lymphocyte (cell/µl)	7029 ± 247	6555 ± 363 (−6.7)###	5203 ± 484 (−25.9)**##	2396 ± 205 (−65.9)***
Monocyte (cell/µl)	184 ± 8	179 ± 14 (−2.7)#	148 ± 17 (−19.5)	133 ± 11 (−27.7)**

^aMean percentage change relative to vehicle control is shown in parentheses. Data shown are mean ± SE (n = 6/group).

Value is significantly different versus vehicle control at *p < 0.05, **p < 0.01 or ***p < 0.001.

Value is significantly different versus positive control (cyclophosphamide) group at #p < 0.05, ##p < 0.01 or ###p < 0.001.

Table 2. Effects of subacute exposure to berberine (BBR, IP daily, 14 days) on mouse spleen cell subtypes.

Parameter	Vehicle	BBR 5 mg/kg^a	BBR 10 mg/kg^a	Cyclophosphamide^a
Spleen weight (mg)	59.6 ± 3.20	60.6 ± 4.40 ( + 1.6)‡	46.2 ± 2.62 (−22.4)*	56.4 ± 4.40 (−5.3)
Cellularity (×10^7)	8.52 ± 1.36	8.16 ± 1.33 (−4.2)	6.12 ± 0.59 (−28.1)	5.77 ± 1.52 (−32.2)
CD19^+ cell (%)	35.89 ± 5.72	27.23 ± 2.33 (−24.1)	18.87 ± 2.10 (−47.4)*	19.80 ± 5.05 (−44.8)*
CD3^+ cell (%)	48.60 ± 3.74	46.60 ± 3.11 (−4.1)	42.00 ± 2.15 (−13.5)	42.75 ± 4.27 (−12.0)
CD4^+ cell (%)	27.55 ± 1.97	31.82 ± 2.42 (+15.4)	26.70 ± 1.71 (−3.0)	29.25 ± 5.19 (+6.1)
CD8^+ cell (%)	17.60 ± 4.40	11.62 ± 0.73 (−33.9)‡#	9.02 ± 0.62 (−48.7)*	8.10 ± 1.75 (−53.9)*
CD4^+:CD8^+ ratio	1.53 ± 0.48	2.73 ± 0.06 (+78.4)‡#*	3.02 ± 0.07 (+97.3)**	3.57 ± 0.37 (+133)***
CD19^+ content (×10^7)	2.85 ± 0.30	2.24 ± 0.26 (−21.4)‡#	1.16 ± 0.17 (−59.2)**	1.00 ± 0.36 (−64.9)**
CD3^+ content (×10^7)	4.28 ± 0.99	3.80 ± 0.63 (−11.2)	2.55 ± 0.12 (−40.4)	2.37 ± 0.62 (−44.6)
CD4^+ content (×10^7)	2.55 ± 0.42	2.59 ± 0.43 (+1.5)‡	1.67 ± 0.12 (−34.5)*	1.51 ± 0.39 (−40.7)
CD8^+ content (×10^7)	1.70 ± 0.69	0.94 ± 0.15 (−44.7)#	0.55 ± 0.03 (−67.6)*	0.43 ± 0.16 (−74.7)*

^aMean percentage change relative to vehicle control is shown in parentheses. Data shown are mean ± SE (n = 6/group).

Value is significantly different versus vehicle control at *p < 0.05, **p < 0.01 or ***p < 0.001.

Value is significantly different versus positive control (cyclophosphamide) group at #p < 0.05.

Value is significantly different as a function of BBR dose at ‡p < 0.05.

Figure 2. Pathologic changes in spleen after administration of 10 mg BBR/kg/day. (a) Vehicle-control group: White pulps with well-formed lymphoid follicles containing active germinal centers. (b) 10 mg BBR/kg/day group: Atrophic white pulps with relatively small lymphoid follicles. Each figure is a representative photomicrograph from a mouse in each group. Magnification = 100×.

Figure 3. Effects of subacute berberine (BBR, 14 days) exposure on mouse antibody response. Levels of anti-SRBC produced in serum harvested from mice 24 h after the final treatment (on Day 15) in indicated regimens. Values shown are mean ± SE (n = 6/group). Value significantly different from vehicle control mice (**p < 0.01 or ***p < 0.001); ##Significantly different versus positive control (CYP) (p < 0.01); or ‡‡significantly different as a function of BBR dose (p < 0.01).

Figure 4. Effects of subacute berberine (BBR, 14 days) exposure on mice DTH response. The percentage increase in paw thickness of mice 24, 48 and 72 h after footpad SRBC challenge in indicated regimens. Values shown are mean ± SE (n = 6/group). Value significantly different from vehicle control mice at *p < 0.05 or **p < 0.01; #significantly different versus positive control (CYP) (p < 0.05); or ‡significantly different as a function of BBR dose (p < 0.05).

Figure 5. Effects of subacute berberine (BBR, 14 days) exposure on ex vivo lymphocyte proliferation. The SI values shown are based upon measures associated with splenocytes harvested from mice 24 h after final treatment (on Day 15) in each regimen. With each treatment group, SI values were calculated from absorbance values for PHA (or LPS)-stimulated cells/absorbance values for unstimulated cells from the same hosts. Values shown are mean ± SE (n = 6/group). Values significantly different from stimulated vehicle control counterpart at *p < 0.05, **p < 0.01 or ***p < 0.001. Value is significantly different versus positive control (CYP) (at #p < 0.05) or as a function of BBR dose (at ‡‡p < 0.01).

Table 3. Effects of subacute exposure to berberine (BBR, IP daily, 14 days) on ex vivo cytokine production by mouse spleen cells.

Parameter	Vehicle	BBR 5 mg/kg^a	BBR 10 mg/kg^a	Cyclophosphamide^a
IFNγ (pg/ml)	22.7 ± 0.5	21.4 ± 0.4 (−5.7)‡‡‡###	15.5 ± 0.6 (−31.7)***	14.7 ± 0.9 (−35.2)***
IL-4 (pg/ml)	8.2 ± 0.3	8.7 ± 0.3 (+6.0)‡‡###	10.5 ± 0.7 (+28.0)**#	11.7 ± 0.3 (+42.6)***

^aMean percentage change relative to vehicle control is shown in parentheses. Data shown are mean ± SE (n = 6/group).

Value is significantly different versus vehicle control at **p < 0.01 or ***p < 0.001.

Value is significantly different versus positive control (cyclophosphamide) group at #p < 0.05 or ###p < 0.001.

Value is significantly different as a function of BBR dose at ‡‡p < 0.01 or ‡‡‡p < 0.001.