The effects of gastric cancer interstitial fluid on tumors based on traditional Chinese medicine 'phlegm' theory and the investigation on the mechanism through microRNA-21 regulation.

This study aimed to investigate the effects of gastric cancer interstitial fluid (GCIF) on tumors and explore the possible mechanism of Xiaotan Sanjie decoction (XTSJ) on treatment of gastric cancer from the view of regulating microRNA-21 (miR-21) expression. The GCIF was extracted and identified by measuring the levels of interleukin-8 (IL-8), intercellular adhesion molecule 1 (ICAM-1) and miR-21. The effects of GCIF on the proliferation of SGC-7901 cells and tumor growing were assessed by cell counting kit-8 (CCK-8) assay and subcutaneously transplanted tumor-bearing nude mice model, respectively. Additionally, inhibition effect of XTSJ decoction on proliferation of SGC-7901 cells intervened by GCIF were assessed in vitro and anti-cancer effect of it was further assessed using orthotopic transplanted tumor-bearing nude mice model. The concentration of SGC-7901 gastric cancer cells were dependent on the concentration of the added GCIF. After 72 hours of continuous culture, the interstitial fluid had an obvious proliferative effect on the SGC-7901 tumor cells, which was the most significant in the high concentration group. XTSJ decoction could inhibit the growth-promoting effect (P < 0.01) of GCIF on gastric cancer cells. Intervention of the GCIF might promote the growth (P < 0.05) of the subcutaneously transplanted tumors in nude mice and decrease the net weight of the tumor-bearing nude mice (P < 0.05) after tumor removal. The GCIF was able to up-regulate the expression (P < 0.001) of miR-21 in the subcutaneously transplanted tumors. XTSJ decoction could downregulate the expression (P < 0.05) of miR-21 in SGC-7901 orthotopically transplanted tumors. XTSJ decoction can inhibit the multiplicative effect of GCIF on gastric cancer cells, growth of gastric tumor and promotion effect of GCIF on tumors, probably due to the down-regulating miR-21 expression in tumor tissues.

Augmentation of hematopoiesis by fibroblast-mediated interleukin-6 gene therapy in mice with chemotherapy.

Murine fibroblast NIH3T3 cells engineered to secrete interleukin-6 (NIH3T3-IL-6) were implanted intraperitoneally into mice and observed for their hematopoietic effects with or without 5-fluorouracil (5-FU) administration. In normal mice, the platelet and neutrophil counts in peripheral blood increased significantly after implantation of NIH3T3-IL-6 cells, but the total white blood cell numbers showed no obvious elevation. The granulocyte-macrophage colony-forming unit (CFU-GM) and megakaryocyte colony-forming unit (CFU-MK) numbers formed by stem cells in bone marrow and spleen were also found to be significantly increased after implantation of NIH3T3-IL-6 cells when compared with those in mice after implantation of NIH3T3 cells transduced with neomycin gene (NIH3T3-Neo). To observe the protective effects of NIH3T3-IL-6 cells on hematopoietic depression in chemotherapy-treated mice, the mice were preinjected with 5-FU at a dosage of 150 mg/kg before implantation of NIH3T3-IL-6 cells. The platelet and neutrophil counts showed accelerated recovery after implantation of NIH3T3-IL-6 cells. The numbers of CFU-GM and CFU-MK in bone marrow and spleen were also found to be markedly increased in hematopoietic-depressed mice when compared with those in mice implanted with NIH3T3-Neo cells. These data suggest that fibroblast-mediated IL-6 gene therapy, which can augment hematopoiesis in mice with or without chemotherapy, will be of great help in the recovery from hematopoietic depression after chemotherapy.

Esculentoside A inhibits tumor necrosis factor, interleukin-1, and interleukin-6 production induced by lipopolysaccharide in mice.

Esculentoside A, a kind of saponin isolated from the root of the Chinese herb Phytolaca esculenta, is reported to possess potent anti-inflammatory effects in acute and chronic experimental models. In the present study, we investigated the effects of esculentoside A on the production of tumor necrosis factor (TNF), interleukin-1 (IL-1) and interleukin-6 (IL-6) induced by lipopolysaccharide (LPS) in mice. In vitro experiments demonstrated that esculentoside A (0.1-10 mumol/l) significantly reduced the release of TNF from the peritoneal macrophages derived from mice pretreated with thioglycolate. IL-1 and IL-6 secretion was also obviously inhibited in a concentration-dependent manner by esculentoside A from 0.01 to 10 mumol/l. In vivo experiments demonstrated that detectable TNF was observed 0.25 h after injection, was maximal at 0.5 h, and returned to baseline at 4 h. Maximal production of IL-1 and IL-6 were observed to be 1 and 2 h, respectively, after injection of LPS. Pretreatment of mice with 5, 10, or 20 mg/kg esculentoside A once a day for 7 consecutive days dose-dependently decreased the TNF, IL-1 and IL-6 levels in the sera of mice following LPS challenge. TNF, IL-1, and IL-6 are important cytokines involved in the pathogenesis of inflammatory lesions. Inhibition of inflammatory cytokine production may contribute to the anti-inflammatory effects of esculentoside A.

Effects of Phytolacca acinosa polysaccharides I on immune function in mice.

Radioactivities of [3H]TdR uptaken by splenocytes and released from [3H]TdR-labeled YAC-1 cell line were measured to determine the degree of lymphocyte proliferation and natural killer (NK) cell activity. Seven days after mice treated with Phytolacca acinosa polysaccharides I (PAP-I) 5-50 mg.kg-1, the NK cell activity, and lymphocyte proliferation induced by Con A 5 micrograms.ml-1 or lipopolysaccharides 10 micrograms.ml-1 were significantly augmented. Splenocytes from mice treated with ip PAP-I 5-50 mg.kg-1 were incubated with Con A 5 micrograms.ml-1 for 24 h to induce interleukin-2 (IL-2) and for 40 h to induce NK cytotoxic factor (NKCF). Radioactivities of [3H]TdR uptaken by CTLL-2 cell line and YAC-1 cell line were used to measure the IL-2 and NKCF activities, respectively. PAP-I enhanced the production of IL-2 and NKCF. These results suggest that PAP-I augments the immunological functions in vivo.

[Effects of Phytolacca acinosa polysaccharide on splenic lymphocyte proliferation and cytokine secretion from splenic lymphocyte and macrophage].

The effects of Phytolacca acinosa polysaccharides I (PAP-I), a polysaccharide extracted from Phytolacca acinosa Roxb: on splenic lymphocyte proliferation and cytokines production from splenic lymphocyte and macrophage were studied. Lipopolysaccharides (LPS) and PAP-I were found to significantly augment splenic lymphocyte proliferation of normal BALB/c, nude BALB/c and NC mice in vitro, but concanavalin A (Con A) was shown to stimulate only normal BALB/c and nude BALB/c splenic lymphocyte proliferation. Also, PAP-I significantly enhanced Con A or LPS-induced lymphocyte proliferation and mixed lymphocyte reaction. Significant enhancement of colony stimulating factor (CSF) production was observed from splenic lymphocyte of normal BALB/c and nude BALB/c mice but not from NC mice when treated with PAP-I for 5 d. PAP-I was shown to significantly enhance interleukin-2 (IL-2) production from normal mice splenocyte and Con A stimulated normal mice splenocyte in a concentration-dependent fashion. Supernatant of PAP-treated macrophage (M phi) were collected and CSF activity was tested. The results confirmed that PAP-I can significantly stimulate M phi to secret CSF activity on d 1. The supernatant also contained a cytokine which exhibited a synergistic action with recombinant murine granular-macrophage CSF (RMGM-CSF) to stimulate mice bone marrow cell proliferation. PAP-I, 5-50 mg.kg-1, ip can enhanced splenic lymphocyte proliferation and IL-2 production. These findings indicate that PAP-I can augment immunologic function in vitro and in vivo.

[Effects of Phytolacca acinosa polysaccharides II on lymphocyte proliferation and colony stimulating factor production from mice splenocytes in vitro].

Phytolacca acinosa polysaccharides II (PAP-II), a kind of polysaccharides isolated from Phytolacca acinosa Roxb with MW 40 kDa, on lymphocyte proliferation and colony stimulating factor (CSF) production from splenocytes in vitro, was studied. The radioactivities of [3H] TdR uptake by lymphocyte and bone marrow cells were used to determine the ability of lymphocyte proliferation and CSF production respectively. PAP-II was found to significantly augment splenocyte proliferation in a dose-dependent fashion, and significantly enhance Con A (1, 2.8 micrograms.ml-1) and LPS (3, 10, 30 micrograms.ml-1) induced lymphocyte proliferation at concentration of 31-125 micrograms.ml-1. As the concentration of PAP-II increased, significant suppression of Con A-induced lymphocyte proliferation was observed. Induction of CSF by PAP-II from splenocyte was confirmed at the present study. The optimal dosage was 100 micrograms.ml-1 and the optimal effect occurred on day 5. These results suggest that PAP-II can augment immunological function and enhance hematopoiesis.