The protective effect of the Vacha rhizome extract on chronic stress-induced immunodeficiency in rat

Figures & data

Table 1. Effects of Vacha rhizome extract on body and lymphoid organ weight of rats.

Groups and treatments	% Change in body weight (g) ± SE	Weight (mg)/100 g body weight mean ± SE
		Spleen	Thymus	Axillary lymph nodes
		Control	13.653 ± 2.276^a	287.666 ± 3.480^b	41.666 ± 3.282^b	48.333 ± 4.910^b
Vehicle control	12.081 ± 0.822^a	283.750 ± 8.910^b	39.000 ± 3.949^b	49.333 ± 1.452^b
Unstressed + benzene extract of Vacha (10 mg/kg body weight)	15.471 ± 1.144^a	310.000 ± 5.773^a	56.333 ± 4.841^a	61.000 ± 1.732^a
Stress	7.289 ± 0.895^b	255.666 ± 6.565^c	24.500 ± 1.936^c	36.333 ± 2.603^c
Stress + benzene extract of Vacha (10 mg/kg body weight)	13.707 ± 0.608^a	286.800 ± 4.115^b	41.400 ± 2.379^b	47.750 ± 3.794^b
ANOVA F Value (df =4, 20)	7.104 p < 0.001	8.023 p < 0.001	9.896 p < 0.001	6.489 p < 0.001

Mean values with same superscript letters in the given column are not significantly different, whereas those with different superscript letters are significantly (p < 0.05) different as judged by Duncan’s multiple test, df: degree of freedom.

Table 2. Effects of Vacha rhizome extract on adrenal 3β-HSDH activity and weight of the adrenal gland of rats.

Groups and treatments	Adrenal 3β-HSDH activity (nmol/mg/min) ± SE	The relative weight of the adrenal gland (mg/100 g of body weight) ± SE
Control	0.452 ± 0.013^a	20 ± 0.001^a
Vehicle Control	0.447 ± 0.031^a	19 ± 0.000^a
Unstressed + benzene extract of Vacha (10 mg/kg body weight)	0.456 ± 0.023^a	19 ± 0.000^a
Stress	0.583 ± 0.044^b	26 ± 0.001^b
Stress + benzene extract of Vacha (10 mg/kg body weight)	0.455 ± 0.028^a	19 ± 0.001^a
ANOVA F Value (df =4, 20)	3.571 p < 0.01	4.159 p < 0.01

Mean values with same superscript letters in the given column are not significantly different, whereas those with different superscript letters are significantly (p < 0.05) different as judged by Duncan’s multiple test, df: degree of freedom.

Table 3. Effects of Vacha rhizome extract on total and differential counts of leukocytes of rats.

Groups and Treatments	Mean total count/μL of leukocytes ± SE	Mean count/μL ± SE
		Lymphocytes	Neutrophils	Monocytes	Basophils	Eosinophils
		Control	4882.000 ± 138.108^b	4168.200 ± 95.876^b	524.250 ± 27.756^b	165.750 ± 15.611^b	65.400 ± 6.523^b	48.500 ± 1.190^b
Vehicle Control	4844.000 ± 267.536^b	3895.200 ± 260.199^b	540.000 ± 21.122^b	169.600 ± 24.254^b	69.500 ± 5.041^b	48.000 ± 3.082^b
Unstressed + benzene extract of Vacha  (10 mg/kg body weight)	5730.000 ± 320.208^a	4832.333 ± 269.499^a	662.000 ± 32.254^a	230.000 ± 15.038^a	96.500 ± 16.500^a	71.000 ± 4.020^a
Stress	3146.000 ± 299.359^c	2512.600 ± 247.264^c	311.250 ± 66.683^c	62.333 ± 2.027^c	35.750 ± 5.022^c	22.750 ± 4.571^c
Stress + benzene extract of Vacha  (10 mg/kg body weight)	4176.666 ± 186.755^b	3591.000 ± 132.061^b	489.250 ± 29.329^b	161.250 ± 8.330^b	69.250 ± 3.520^b	45.250 ± 3.567^b
ANOVA F Value (df =4, 20)	13.844 p < 0.001	15.536 p < 0.001	9.536 p < 0.001	11.638 p < 0.001	8.338 p < 0.001	22.538 p < 0.001

Mean values with same superscript letters in the given column are not significantly different, whereas those with different superscript letters are significantly (p < 0.05) different as judged by Duncan’s multiple test, df: degree of freedom.

Table 4. Effects of Vacha rhizome extract on circulating immune complexes and total immunoglobulins contents of rats.

Groups and treatments	Total immunoglobulins Content (mg/mL) ± SE	Circulating immune complexes PEG index ± SE
Control	0.542 ± 0.009^b	78.600 ± 2.249^b
Vehicle Control	0.537 ± 0.044^b	78.000 ± 2.915^b
Unstressed + benzene extract of Vacha (10 mg/kg body weight)	0.645 ± 0.045^a	44.200 ± 2.745^c
Stress	0.372 ± 0.023^c	126.000 ± 2.983^a
Stress + benzene extract of Vacha (10 mg/kg body weight)	0.499 ± 0.028^b	75.800 ± 3.382^b
ANOVA F Value (df = 4, 20)	10.389 p < 0.001	113.690 p < 0.001

Mean values with same superscript letters in the given column are not significantly different, whereas those with different superscript letters are significantly (p < 0.05) different as judged by Duncan’s multiple test, df: degree of freedom.

Table 5. Effects of Vacha rhizome extract on bone marrow stem cells of rats.

Groups and treatments	The number of bone marrow stem cells (×10^4/mL) ± SE	Viability of bone marrow stem cells (%) ± SE
Control	773.715 ± 8.699^b	96.200 ± 0.860^a
Vehicle control	753.124 ± 7.270^b	93.800 ± 0.860^a
Unstressed + benzene extract of Vacha (10 mg/kg body weight)	853.590 ± 48.451^a	96.000 ± 1.000^a
Stress	664.862 ± 17.344^c	86.800 ± 1.392^b
Stress + benzene extract of Vacha (10 mg/kg body weight)	738.906 ± 14.835^b	93.000 ± 0.707^a
ANOVA F Value (df = 4, 20)	8.713 p < 0.001	14.027 p < 0.001

Mean values with same superscript letters in the given column are not significantly different, whereas those with different superscript letters are significantly (p < 0.05) different as judged by Duncan’s multiple test, df: degree of freedom.

Table 6. Effects of Vacha rhizome extract on lymphocyte counts in different lymphoid organs of rats.

Groups and treatments	The number of lymphocytes (× 10^6/mL) ± SE
	Spleen	Thymus	Axillary lymph nodes
	Control	188.00 ± 2.213^b	1530.400 ± 2.227^b	754.200 ± 3.624^b
Vehicle control	179.000 ± 5.431^b	1527.000 ± 3.114^b	743.400 ± 1.964^b
Unstressed + benzene extract of Vacha (10 mg/kg body weight)	229.800 ± 12.276^a	1724.400 ± 11.732^a	799.800 ± 7.009^a
Stress	142.000 ± 16.077^c	1405.400 ± 6.874^c	545.600 ± 25.085^c
Stress + benzene extract of Vacha (10 mg/kg body weight)	174.000 ± 2.645^b	1510.333 ± 2.603^b	726.200 ± 3.611^b
ANOVA F Value (df =4, 20)	44.532 p < 0.001	281.433 p < 0.001	67.900 p < 0.001

Mean values with same superscript letters in the given column are not significantly different, whereas those with different superscript letters are significantly (p < 0.05) different as judged by Duncan’s multiple test, df: degree of freedom.

Figure 1. (a–e) Photomicrographs of splenocytes stained with acridine orange and ethidium bromide (450 nm and 530 nm). Note the presence of more number of healthy cells (N) in control (a), vehicle control (b), unstressed + Vacha extract treated rats (c) and stress + Vacha extract treated rats (e), and more number of apoptotic cells (A) in stressed (d) rats. 200×. A: apoptotic cells; N: normal, healthy cells.

Table 7. Effects of Vacha rhizome extract on apoptosis of cells in lymphoid organs of rats.

Groups and treatments	Apoptotic index (%)
	Spleen	Thymus	Axillary lymph nodes
	Control	10.600 ± 1.029^b	12.400 ± 0.927^b	14.400 ± 1.208^b
Vehicle Control	11.000 ± 1.581^b	12.000 ± 1.949^b	12.800 ± 1.881^b
Unstressed + benzene extract of Vacha (10 mg/kg body weight)	9.200 ± 0.663^b	10.800 ± 0.734^b	11.800 ± 1.067^b
Stress	25.000 ± 1.843^a	29.000 ± 2.738^a	31.200 ± 3.455^a
Stress + benzene extract of Vacha (10 mg/kg body weight)	11.800 ± 1.593^b	13.200 ± 1.280^b	18.400 ± 1.208^b
ANOVA F Value (df =4, 20)	21.163 p < 0.001	20.179 p < 0.001	16.149 p < 0.001

Mean values with same superscript letters in the given column are not significantly different, whereas those with different superscript letters are significantly (p < 0.05) different as judged by Duncan’s multiple test, df: degree of freedom.

Figure 2. (a–e) Photomicrographs of the cross sections of the spleen showing the island of white pulp. Note the shrinkage of island of the white pulp region and germinal center in stressed rats (d) compared to control (a), vehicle control (b), unstressed + Vacha extract (c) and stress + Vacha extract (e) treated rats. 200× (H&E). GC: germinal center; CA: central artery; L: lymphocytes.

Table 8. Effects of Vacha rhizome extract on number of islands of white pulps in spleen of rats.

Groups and treatments	Number of islands of white pulp/cross section of spleen ± SE
Control	27.500 ± 1.322^b
Vehicle Control	27.250 ± 1.108^b
Unstressed + benzene extract of Vacha (10 mg/kg body weight)	34.200 ± 1.772^a
Stress	21.500 ± 1.322^c
Stress + benzene extract of Vacha (10 mg/kg body weight)	26.600 ± 1.326^b
ANOVA F Value (df =4, 20)	10.217 p < 0.001

Mean values with same superscript letters in the given column are not significantly different, whereas those with different superscript letters are significantly (p < 0.05) different as judged by Duncan’s multiple test, df: degree of freedom.

Figure 3. (a–e) Photomicrographs of the cross sections of thymus gland. Note the reduction of cortex region and development of connective tissue and fat cells in stressed (d) rats compared to control (a), vehicle control (b), unstressed + Vacha extract (c) and stress + Vacha extract (e) treated rats. 200× (H&E). C: connective tissue fibers; F: fat cells.

Figure 4. (a–e) Photomicrographs of cross sections of axillary lymph node showing lymphoid follicles. Note the shrinkage of germinal centers in stressed (d) rat compared to control (a), vehicle control (b), unstressed + Vacha extract (c) and stress + Vacha extract (e) treated rats. 200× H&E. GC: germinal centre; L: B lymphocytes.