Aim To investigate the effects of ursolic acid from loquat leaves on proliferation inhibition and expression of PPAR-γ 、TGF-β1 in rat hepatic stellate cells, so as to explore the mechanism of anti-hepatic fibrosis of UA.Methods HSC-T6 cells were randomly divided into blank group, rosiglitazone control group, the low, medium and high concentration of UA group to detect the cell proliferation inhibition by CCK-8 after 24,48,72 h.The content of type collagen Ⅰ in cell culture supernatant of each group was examined by ELISA.The expression of PPAR-γ mRNA, TGF-β1 mRNA in HSC-T6 cells exposured were examined by real-time quantitative PCR.Effects on HSC-T6 PPAR-γ and TGF-β1 protein from each group were detected by immunocytochemical method.Results CCK-8 results showed that the inhibitory rate of UA on cell proliferation increased with the prolongation of drug action time(P<0.01).ELISA results showed that with the increase of the concentration of UA, the content of type Ⅰ collagen content decreased(P<0.01).Real-time PCR results showed that with the increase of the concentration of UA, and the expression of PPAR-γ mRNA increased, the expression of TGF-β1 mRNA decreased(P<0.01).The results of immunocytochemistry showed that the expression of PPAR-γ protein was increased, and the expression of TGF-β1 protein was decreased with the increase of the concentration of UA(P<0.01).All effects mentioned above were dose-dependent.Moreover, the effects in the high concentration groups were stronger than those in control group.Conclusion UA can inhibit the proliferation of HSC-T6 cells, Which may be associated with the up-regulation of PPAR-γ expression and the down-regulation of TGF-β1 expression.

Objective: To investigate the enhancement effect of Xinjiang wild Artemisia rupestris L. crude polysaccharides (WARCP) as an adjuvant on ovalbumin (OVA) vaccine in mice immunized intramuscularly. Methods: ICR mice were randomly divided into 6 groups (5 per group), including 9 g/L NaCl group (blank control), OVA group (10 μg OVA), low dose WARCP/OVA group (OVA+50 μg WARCP), medium dose WARCP/OVA group (OVA+200 μg WARCP), high dose WARCP/OVA group (OVA+400 μg WARCP), and aluminum adjuvant (Alum)/OVA group (positive control group, OVA+100 μg Alum). ICR mice were immunized intramuscularly and weighted. The OVA-specific antibodies and subtypes in serum were detected by enzyme linked immunosorbent assay (ELISA). T cells subsets from spleen and lymph nodes were detected by flow cytometry. Results: The medium-dose WARCP/OVA group enhanced IgG and IgG1 levels and increased early antibody levels (all P<0.05). The medium-dose WARCP/OVA group and the high-dose WARCP/OVA group significantly enhanced IgG2a levels (all P<0.05), but the difference was not statistically significant comparing with Alum/OVA group (P>0.05). The low-dose WARCP/OVA group enhanced the percentage of CD4⁺ T cells in spleen and CD4⁺ T, CD8⁺ T, CD4⁺CD44⁺ T cells in lymph nodes (all P<0.05). The medium dose WARCP/OVA group and the high dose WARCP/OVA group enhanced the CD4⁺ T, CD8⁺ T, CD4⁺CD44⁺ T, CD8⁺CD44⁺ T cells in spleen and CD8⁺CD44⁺ T cell in lymph nodes (all P<0.05). Conclusions Plant-derived WARCP as an OVA protein vaccine adjuvant can enhance cellular immunity and humoral immunity, and it is safe and reliable. The results in this study provide a theoretical basis for the popularization and application of WARCP.

Objective To compare the immunopotentiating effects of polysaccharides extracted from wild/cultivate Cistanehe deserticola (WCDPS/CCDPS) in Xinjiang. Methods ICR mice were subcu-taneously injected twice with different doses(low,medium and high) of WCDPS and CCDPS in combination with ovalbumin (OVA). OVA-specific antibody IgG,as well as IgG1 and IgG2a subtypes, was determined by ELISA. OVA-specific lymphocyte proliferation was measured by MTT. Expression of CD4+T and CD8+T cells was analyzed by flow cytometry. Results Both WCDPS and CCDPS could significantly improve the production of OVA-specific IgG,IgG1 and IgG2a,promote the proliferation of OVA-specific lymphocytes and increase the expression of CD4+T and CD8+T cells(all P<0.05) with no significant difference between them at the same dosages (P>0.05). WCDPS and CCDPS had no influence on the body weight of mice after im-munization. Conclusion WCDSP and CCDPS could significantly enhance the OVA-specific humoral and cellular immune responses with no statistical difference and are characterized by high safety.

Objective To study the effects of aqueous extracts of cultivated Cistanche deserticola Y. C. Ma (AECCD) on T cell responses and the duration of antibody response and to investigate its immunoen-hancing activities in mice. Methods Two batches of female ICR mice were used in this study with 30 from each batch. Each batch of mice was randomly divided into six groups (n=5). Low, medium and high doses of AECCD in combination with ovalbumin ( OVA) were used to set up three experimental groups, while 0. 9% NaCl, OVA alone and aluminium adjuvant were respectively used as blank, negative and positive controls. All mice were intramuscularly injected twice at an interval of two weeks. Flow cytometry was used to detect the ex-pression of T lymphocyte subsets, cytokines and surface molecules of dendritic cells (DC). Indirect ELISA was used to detect IgG antibody levels. Results AECCD could significantly increase the percentage of CD4+and CD8+T lymphocytes in spleen (P<0. 05), up-regulate the expression of CD4+CD44+and CD8+CD44+effector T lymphocytes (P<0. 05), promote the secretion of IFN-γ in T lymphocytes and enhance the expression of CD40 and CD80 on the surface of DC (P<0. 05). ELISA results showed that high-dose AECCD could significantly prolong the duration of IgG antibody response induced by OVA (P<0. 05). Conclusion AECCD could en-hance the T lymphocyte immune response induced by OVA and keep it maintained at a high level, which might help to improve the body′s immune response.

Objective:To investigate the effect of wild Cistanche deserticola crude polysaccharides (WCDCP) on the immune response of ovalbumin (OVA).Methods:42 ICR mice were randomly divided into the 9 g/L NaCl group (blank sample), WCDCP group (400 μg WCDCP), OVA group (10 μg OVA), low-dose WCDCP/OVA group (100 μg WCDCP+10 μg OVA), medium-dose WCDCP/OVA group (400 μg WCDCP+10 μg OVA), high-dose WCDCP/OVA group (800 μg WCDCP+10 μg OVA), and aluminum adjuvant/OVA group (positive concentration, 200 μg aluminum adjuvant+10 μg OVA). Each group included 6 mice. The mice were immunized by using two-point injection into the muscles of the hind legs of the mice. A total of 2 immunizations, and the immunization was boosted once every 2 weeks after the initial immunization. The body weight of the mice was weighed 7, 14, 21, and 28 days after the initial immunization of the mice, and the changes in body weight and growth status of the mice were observed. The IgG antibodies and antibody fractions were detected by indirect enzyme-linked immunosorbent assay. Twenty-one days after the initial immunization, the spleen lymphocyte proliferation level was detected by the thiazole blue method. Flow cytometry was used to detect the proportion of T cell subsets in the spleen and lymph nodes.Results:At 7 days after the initial immunization, the serum IgG antibody level (0.597 6±0.110 7) in the high-dose WCDCP/OVA group was significantly higher than (0.254 4±0.074 8) of the OVA group ( P<0.05). At 28 days after the initial immunization, the serum IgG, IgG1 and IgG2a antibody levels in the high-dose WCDCP/OVA group were higher than those in the OVA group, the comparison respectively were 0.972 3±0.243 8 vs. 0.389 2±0.077 4 ( P<0.05), 1.156 0±0.088 4 vs. 0.612 6±0.059 7 ( P<0.001), 1.648 0±0.103 9 vs. 0.557 2±0.181 5 ( P<0.001), and the differences were statistically significant. High-dose WCDCP can significantly promote the proliferation of spleen cells induced by concanavalin A ( P<0.001) and lipopolysaccharide ( P<0.05). High-dose WCDCP/OVA group can significantly stimulate the activation ratio of T cell subsets in the spleen. The proportion of CD3 +CD8 + T cells in the high-dose WCDCP/OVA group [(10.83±0.44)%] was significantly higher than that in the OVA group[(6.76±0.58)%] ( P<0.01), and the proportion of CD3 +CD4 + T cells [(28.17±1.67)%] was also significantly higher than that in the OVA group [(19.17±2.73)%] ( P<0.05). WCDCP had no effect on body weight (all P>0.05) and growth of mice. Conclusions:WCDCP can enhance humoral immune response and cellular immune response, and has no side effect on the growth of mice, suggesting that WCDCP can be used as a potential adjuvant for OVA.