Echinacea: The genus Echinacea

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Two Immunoenhancers Are Not Better than One 223 identified and recorded from 2,000 total cells (hemopoietic/immune) per organ (spleen, bone marrow) per mouse, the absolute numbers of each of these cell types could be calculated from the already determined total organ cellularity. This latter value was obtained previously for each spleen and each femur by means of the electronic cell counter at the time of animal killing. Finally, the data were analyzed statistically by means of the student’s t-test (two-tailed). Thus, the means of experimental and control groups of mice were calculated. Via the t-test, the differences were ascertained between the means of the various groups: MLT versus MLT + E. purpurea; MLT versus control chow; and MLT + E. purpurea versus control chow. This was done for each of six cell subgroups, for each organ (spleen, bone marrow) for each mouse in each of the three diets. RESULTS AND INTERPRETATIONS Fourteen days of in vivo administration of MLT alone versus the control diet had no statistically significant effect on the absolute numbers of nucleated erythroid cells (precursors of blood-borne red blood cells) in the spleen (Figure 16.1a) or the bone marrow (Figure 16.1b). These cells in the spleen were the late erythroid precursors, that is, small, postmitotic, possessing a very dark nucleus indicative of impending pycnosis, and a cytoplasm showing clear evidence of hemoglobin. However, they were significantly reduced in the spleens in mice administered both MLT + E. purpurea, compared to either the control diet, or MLT only groups (Figure 16.1a). A strikingly different result was obtained in the bone marrow, however, with regard to cells of the erythroid lineage (Figure 16.1b). In the bone marrow, the nucleated erythroid cells, unlike the spleen, are predominantly large, proliferating, and show no cytoplasmic hemoglobin. In this organ, the absolute numbers of these nucleated cells was significantly elevated (Figure 16.1b) in mice receiving MLT + E. purpurea versus either MLT only or control vehicle diet. Cells of the granulocytic lineage, including both mature, functional forms (Figure 16.2a and Figure 16.2b), and their proliferating precursor forms (Figure 16.3a and Figure 16.3b), in both the spleen and bone marrow, followed a very similar response pattern when mice were given both MLT and E. purpurea simultaneously in the diet. That is, with respect to the functionally mature cells of this lineage (residents of both the spleen and bone marrow), MLT alone had little influence on their numbers, relative to the control diet group (Figure 16.2a and Figure 16.2b). However, when mice were fed MLT + E. purpurea, these cells became profoundly reduced in numbers in the spleen compared to the control diet group or the MLT only group (Figure 16.2a). Similarly, in the bone marrow, mature granulocytic cells were significantly reduced relative to mice fed MLT only or the control diet (Figure 16.2b). By contrast, immature (early proliferating precursor) cells in this lineage were significantly elevated in absolute numbers, in both the spleen and bone marrow of mice given MLT + E. purpurea. These immature cells were found in numbers 28-fold and 7-fold, in the spleen and bone marrow, respectively, of the values found in mice given the control diet (Figure 16.3a and Figure 16.3b). In this study, therefore, we found little or no influence of MLT, administered alone, on erythroid, granulocytic cells in either organ. We have previously observed, moreover, that the herb E. purpurea, administered in the diet by itself had no influence on the erythroid and granulocytic cell lineages, when administered for either 7 or 14 days (Sun et al., 1999). From evidence in vitro, MLT appears to stimulate GM-CFU indirectly, acting through MLT receptors on bone marrow stromal cells, the latter producing subsequently a variety of hemopoiesis-driving cytokines (Maestroni, 1998; Maestroni et al., 1994a, 1994b). In accord with these findings, we found in the present in vivo study, a significant increase in mice consuming MLT only, of proliferating precursors of the granulocytic lineage in the bone marrow, but not in the spleen (Figure 16.3b: MLT versus vehicle). The effect of co-administration of MLT and E. purpurea appears to be directly correlated with the level of cell maturity within both lineages (erythroid and granulocytic). That is, mature cells are significantly reduced in number in both organs (spleen, bone marrow) while the early, prolif- © 2004 by CRC Press LLC
224 Echinacea: The genus Echinacea (×10 6 ) (×10 6 ) 25 20 15 10 5 0 MLT MLT + E.p. Vehicle FIGURE 16.1 Absolute numbers of nucleated erythroid cells in the spleen (a) and bone marrow (b) of untreated (control) mice, mice given melatonin (MLT) alone, or mice simultaneously administered MLT + E. purpurea (E.p.), daily for 14 days. Mean ± standard error: 9 to 10 mice/group. p < 0.02 (MLT vs. MLT + E.p.: spleen); p < 0.001 (control vs. MLT + E.p.: spleen); p < 0.02 (MLT vs. MLT + E.p.: bone marrow); p < 0.04 (control vs. MLT + E.p.: bone marrow). erating precursors are significantly elevated in number. These observations suggest that maturation has been inhibited in the presence of both agents administered together, but not separately (Currier et al., 2000; Sun et al., 1999). An anti-apoptotic activity has already been ascribed to MLT (Maestroni, 1998; Provinciali et al., 1996; Yu et al., 2000), and it is possible that this effect is enhanced, additively or synergistically, by the presence of some as yet unidentified component in E. purpurea, resulting in the observed accumulation of the early erythroid- and granulocyticproliferating precursors, concomitant with a striking paucity of their mature progeny. E. purpurea, however, when administered alone, appears to have no anti-apoptotic characteristics, according to our previous observations of unchanged numbers of all cells (mature and precursor) in these two major hemopoietic cells lineages during either 7 or 14 days of dietary administration (Sun et al., 1999). Inhibition of maturation in these two vital cell lineages (erythroid and granulocytic) is clearly undesirable, given that the mature cells in each lineage are the functional ones. A halt in development © 2004 by CRC Press LLC 3 2.5 2 1.5 1 0.5 0 MLT MLT + E.p. Vehicle (a) (b)
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Echinacea The genus Echinacea © 20
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Echinacea The genus Echinacea Edite
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Contents Series Preface Preface Con
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Sandra C. Miller Chapter 17 Adverse
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Multinational pharmaceutical compan
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as tissue retention/distribution, b
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David D. Kitts Food, Nutrition and
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Section I The Genus Echinacea: Taxo
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4 Echinacea: The genus Echinacea HI
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10 Echinacea: The genus Echinacea A
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2 A Review of Preliminary Echinacea
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A Review of Preliminary Echinacea G
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Section II Cultivation of Genus Ech
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© 2004 by CRC Press LLC TABLE 4.6
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5 Introduction Nitrogen Compounds A
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TABLE 5.1 Alkamides from Echinacea
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Phytochemistry of the Genus Echinac
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Section IV Analytical Evaluation in
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8 Factors Affecting Echinacea Quali
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Factors Affecting Echinacea Quality
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© 2004 by CRC Press LLC TABLE 8.1
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9 Popularity, Diversity, and Qualit
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Popularity, Diversity, and Quality
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Echinacoside Echinacoside is a poly
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TABLE 9.13 Microbiological Testing
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Berti, M., Wilckens, R., Fischer, S
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Kreis, W., Sussner, U. and Nusslein
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U.S. Pharmacopeia, 2002, The Standa
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