| In this paper, the modulation effects and mechanism of action of TCA on functions of immunocyte were studied by observing the effects of TCA on macrophage and T, B lymphocyte in mice. The main research contents as follows:In order to illuminate the effects of TCA on phagocytosis of macrophage in mice, the CRBC semi-intracorporal method and toluylene red phagocytose method were adopted to determine the effect of TCA on phagocytosis of macrophage in mice in vivo and vitro.The results showed that TCA could remarkably increase the percentage of the peritoneal macrophage phagocytizing CRBC in mice (P<0.01), remarkably increase its cytophagic index (P<0.05).TCA at 0.015, 0.3, 1.5 and 15μg/mL can remarkably increase the phagotrophic rate of peritoneal macrophage cultured in vitro phagocytizing toluylene red (P<0.05). It indicated that TCA could raise the phagocytosis of macrophage. The effects of TCA on macrophage of mice for secreting IL-1βwas determined by ELISA, the results showed that TCA at low and high dose both could increase the content of IL-1βin blood serum of normal and immune suppression mice(P<0.01), TCA was administratered at 1.5 and 15μg/mL could remarkably increase the IL-1βsecretory volume of mice peritoneal macrophage cultured in vitro(P<0.01), TCA was administratered at 0.015, 1.5, 0.3 and 15μg/mL could remarkably increase the volume of IL-1βwhich was secreted by peritoneal macrophage of mice activated by LPS cultured in vitro (P<0.05),. It indicated that TCA had the promotive effects on macrophage of mice for secreting IL-1β. The content of TNF-αin blood serum of normal and immunosuppressive mice were detected by ELISA, the results showed that TCA could increase the content of IL-1βin blood serum of normal and immune suppressive mice remarkably(P<0.01).This indicated that TCA had stimulative effect on peritoneal macrophage secreting TNF-α..The effects of TCA on the proliferative response of normal peritoneal macrophage and peritoneal macrophage stimulated by LPS and CsA at q.s were detected by MTT method,The results showed that TCA could remarkably increase the proliferative activity of mice peritoneal macrophage (P<0.01), and its effects reached the peak after medication 12h, TCA at 0.3μg/mL could remarkably increase the proliferative response of peritoneal macrophage stimulated by LPS at q.s (P<0.05), TCA was administratered at 0.015, 0.3, 1.5 and 15μg/mL had remarkable recovery effects on the proliferative response of peritoneal macrophage suppressed by CsA (P<0.05). It indicated that TCA had stimulative effect on the multiplication and differentiation of macrophage.To elucidate the modulating effects and mechanism of action of TCA on the function of T and B lymphocyte, a MTT method was adopted to determine the effects of TCA on the proliferative response of normal splenic lymphocyte and it stimulated by LPS, ConA and CsA cultured in vitro,The results showed that proliferative activity of splenic lymphocyte was increased gradually with the increasing of concentration of TCA, this effect could reach the peak at 24h after adding medicine, TCA was administratered at 15μg/mL could remarkably increase proliferative activity of splenic lymphocyte cultured in vitro(P<0.05), remarkably increase proliferative activity of splenic lymphocyte activated by LPS cultured in vitro(P<0.01), remarkably increase proliferative activity of splenic lymphocyte suppressed by CsA cultured in vitro(P<0.01). It indicated that TCA had promotional effect on proliferative activity of splenic lymphocyte of mice.The effects of TCA on T, B lymphocyte and its subpopulation in peripheral blood and spleen of normal and immunosuppressive mice were detected by flow cytometry, The results showed that TCA at high and low dose both could notablely increase CD3+, CD4+, CD8+ cell and ratio of CD4+cell/CD8+ cell in peripheral blood of mice(P<0.01), TCA at low dose could notablely increase CD19+ cell in peripheral blood and spleen of mice(P<0.01), TCA at high dose could notablely recover percentage of CD3+ cell in peripheral blood and spleen of mice suppressed by CsA(P<0.01), TCA at low dose could notablely recover percentage of CD19+ cell in peripheral blood and spleen of mice suppressed by CsA(P<0.01), TCA at high and low dose both could notablely recover percentage of CD4+, CD8+ cell and the ratio of CD4+cell/CD8+ cell in peripheral blood of mice suppressed by CsA(P<0.01), TCA at high and low dose both can increase the percentage of CD3+ in spleen of mice(P<0.01). It indicated that TCA has the effects of leading the increase of Th and Ti cell and the decrease of Ts cell on the body. The content of IgG in blood serum of normal and immunosuppressive mice and the supernatant of splenic lymphocyte of mice cultured in vitro was detected by ELISA, the result showed that TCA could increase the content of IgG in blood serum of normal mice remarkably(P<0.05),could notablely increase the volume of IgG which was secreted by splenic lymphocyte of mice cultured in vitro(P<0.05), notablely recover the volume of IgG which was secreted by splenic lymphocyte of mice suppressed by CsA cultured in vitro(P<0.05), notablely increase the volume of IgG which was secreted by splenic lymphocyte of mice stimulated by LPS of mice cultured in vitro(P<0.05). It indicated that TCA had the effect of promoting B lymphocyte to secrete IgG.. The relative expression of IL-2 gene in the spleen of normal and immunosuppressive mice and the normal splenic lymphocyte and it stimulated by LPS and ConA cultured in vitro was determined by real-time fluorescent quantitation RT-PCR, and an ELISA was adopted to detect the effects of TCA on the normal splenic lymphocyte and it stimulated by LPS, ConA and CsA at q.s. secreting IL-2, the results showed that TCA was administratered at low dose could remarkably increase the expression of IL-2 mRNA in spleen of immunosuppressive mice(P<0.01), but can decrease the expression of IL-2 mRNA in mice splenic lymphocyte cultured in vitro(p<0.01), TCA was administratered at different dose both had the promotional effect on splenic lymphocyte cultured in vitro secreting IL-2 protein and presented a dose-dependent relationship. TCA could notablely increase the volume of IL-2 which was secreted by of splenic lymphocyte activated by LPS and ConA(P<0.05), increase the volume of IL-2 which was secreted by splenic lymphocyte suppressed by CsA significantly(P<0.05).Conclusions: the accommodative effect of TCA on nonspecific immune function was carried out by enhancing phagocytosis and proliferative response and secretion function of macrophage, or by recovering and promoting the macrophage to secrete the IL-1βand TNF-α; The accommodative effect of TCA on nonspecific immune function of mice was relative to its effect of increasing and recovering the proliferation and secretion function of T, B lymphocyte in peripheral blood and spleen of mice.
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