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Polyprezinc up-regulates IGF-I mRNA expression (1)

Polyprezinc up-regulates IGF-I mRNA expression (1)

  • Categories:Stomach healthy
  • Author:
  • Origin:
  • Time of issue:2020-12-03
  • Views:0

(Summary description)

Polyprezinc up-regulates IGF-I mRNA expression (1)

(Summary description)

  • Categories:Stomach healthy
  • Author:
  • Origin:
  • Time of issue:2020-12-03
  • Views:0
Information

  Preface

  Insulin-like growth factor I (IGF-I) is a polypeptide similar in structure to proinsulin. It plays an important role in cell proliferation, differentiation, and glucose uptake. However, the expression of IGF-I gene is affected by many factors. Such as growth hormone, estrogen and cAMP response factor. Zinc is an essential trace element for the human body and a component of enzymes and transcription factors. It has many biological functions such as nucleotide synthesis, protein synthesis and gene expression during cell proliferation and differentiation. Zinc deficiency can hinder human growth and development and ulcer healing, leading to various ulcer diseases. Studies have shown that zinc deficiency can reduce the serum IGF-I concentration, and zinc supplementation can reverse this change.

  Polyprezinc (Polaprezinc, PZ) is a chelate composed of zinc and L-carnosine, and is often used clinically to treat gastric ulcers. Studies have shown that Polyprezinc can reverse the delayed healing of gastric ulcers caused by the lack of zinc. From this point of view, zinc plays a vital role in ulcer healing. So by what way does Polyprezinc promote ulcer healing? What role does IGF-I play in it? Look at the following research.

  Materials and Method

  1.1 Cell culture

  Human umbilical vein endothelial cells (HUVEC) are cultured in E-GM ultraviolet medium; foreskin fibroblasts are derived from in vitro cells and cultured in 106 medium. The guinea pig gastric mucosal epithelial cells were prepared by collagenase digestion.

  1.2 Cell proliferation

  The cells were cultured for several days. After washing the cells with MEM without fetal bovine serum, polyprezinc and other reagents containing or not containing 3% serum were added to the culture dishes, and cultured for 21 hours to evaluate the intake of BrdU. cell counts. Cell proliferation ELISA and BrdU calorimetry kit were used to analyze BrdU intake. After the cells were cultured for 21 hours, they were incubated with BrdU for 3 hours, the medium was removed, and the cells were fixed with ethanol. After removing the ethanol, add anti-BrdU antibody to each well and incubate for 1 h. After the antibody solution is removed, the cells are washed and incubated in the substrate solution for 15 min. Stop the reaction with 1NH2SO4, measure the absorbance at 450nm, and count the cells with a hemocytometer.

  To evaluate the time course of BrdU uptake, cells were washed with MEM and cultured in MEM for 24 hours. Polyprezinc was added to the conditioned medium, the cell culture time is shown in Figure 2, and the BrdU uptake was analyzed for 3 hours.

  In the experiment of anti-IGF-I antibody, HUVEC was incubated with polyprezinc or anti-IGF-I antibody in MEM for 21h, and then BrdU was added to the medium. After the cells were cultured for 3 hours, the BrdU uptake was measured.

  1.3 IGF-I gene expression

  The cells were cultured in a 75cm² tissue culture flask containing MEM for 24h, polyprezinc was added to the medium, and the cells were cultured within the time shown in Figure 4. The medium was removed, the cells were washed with PBS, and the total RNA of the cells was separated by the AGPC method. The content of IGF-I mRNA was detected by rTth-DNA polymerase and Southern hybridization one-step PCR method. Amplify total RNA. The signal on the X-ray film was detected with the ECL detection kit, the antibody was labeled with anti-fluorescein horseradish peroxide, and the X-ray film was analyzed with the V10 image analyzer.

  1.4 Statistical analysis

  The results are expressed as mean ± SE, and Dunnett's test was used for data analysis.

  Results and analysis

  2.1 The effect of polyprezinc on cell proliferation

  First, we investigated whether polyprezinc has the ability to stimulate cell proliferation. The addition of polyprezinc (final concentration of 3×10-10-3×10-8M) in low serum medium to guinea pig gastric mucosal epithelial cells has no effect on BrdU intake and cell number. However, polyprezinc gradually increased the number of HUVEC and foreskin fibroblasts (Table 1). Polyprezinc does not affect the growth of rabbit gastric mucosal epithelial cells, MKN45 cells and human gastric cancer cells. It shows that polyprezinc can promote the proliferation of non-parenchymal cells such as endothelial cells and fibroblasts, but has no obvious effect on epithelial cells.

  Table 1 The effect of PZ on cell proliferation

  2.2 The effect of polyprezinc and its components on cell proliferation

  In order to determine the important components of polyprezinc to promote cell proliferation, we used zinc sulfate and L-carnosine for research. In HUVEC, zinc sulfate (10-8M) increases the intake of BrdU under serum-free conditions, but the effect of zinc sulfate is weaker than that of polyprezinc; when L-carnosine alone exists, the addition of zinc-free medium does not Change the intake of BrdU (Figure 1). It shows that the effect of polyprezinc on promoting cell proliferation is mainly attributed to zinc ions.

  Figure 1 The effect of PZ and its components on the uptake of HUVEC BrdU

  2.3 The effect of polyprezinc on the time course of BrdU intake

  After 8 hours of adding polyprezinc to the medium without zinc or serum, the intake of BrdU did not change, but the intake of BrdU began to increase after 16 hours and after the addition of serum (Figure 2).

  Figure 2 Time course changes of PZ to HUVEC BrdU intake

  2.4 The effect of polyprezinc on IGF-I gene expression

  In HUVEC, the content of IGF-I mRNA increased with the concentration of polyprezinc (Figures 3 and 4). The PCR product of IGF-I has two different lengths (345bp and 420bp) on PAGE (Figure 3).

  Figure 3 The effect of PZ on IGF-I gene expression

  Figure 4 The effect of PZ on IGF-I gene expression

  2.5 The effect of polyprezinc on the time change of IGF-I gene expression

  Both products were identified by Southern blotting of IGF-I cDNA (Figure 5a). In addition, 3h after adding the drug, the IGF-I mRNA content increased, and then gradually decreased until 24h (Figure 5a), but the β-actin mRNA content did not change (Figure 5b). The mRNA levels of other growth factors, such as human growth factor, epidermal growth factor and TGF-β, were also unchanged.

  Figure 5 Time changes of PZ on IGF-I gene expression

  2.6 The effect of polyprezinc on the time change of IGF-I gene expression

  Anti-human IGF-I antibody was used to study the effect of IGF-I on the increase of BrdU intake caused by polyprezinc.

  This antibody is specific for human IGF-I, but does not recognize IGF-II. Figure 6 shows that polyprezinc combined with anti-IGF-I antibody significantly inhibits the uptake of BrdU by HUVEC. However, it is not affected by normal immunoglobulin G.

  Figure 6 The effect of IGF-I on HUVEC BrdU intake by PZ

  Discuss

  Polyprezinc promotes the proliferation of non-parenchymal cells such as endothelial cells and fibroblasts to promote the healing of ulcers and ulcers. This effect is mainly attributed to zinc ions. Polyprezinc has higher adhesion and permeability to gastric mucosa than zinc sulfate, and its anti-ulcer effect is also stronger than zinc sulfate, indicating that in polyprezinc, L-carnosine enhances the effect and absorption of zinc ions.

  IGF-I receptor is expressed by gastric epithelial cells, and IGF-I promotes the proliferation of gastric epithelial cells. Polyprezinc can promote the expression of IGF-I gene, thereby increasing the intake of BrdU. When polyprezinc is combined with anti-IGF-I antibody, it will significantly inhibit the intake of BrdU by HUVEC, indicating that IGF-I is involved. In this way, polyprezinc and IGF-I both play a crucial role in the proliferation of cells.

  Boda Weiye Ruilaisheng (Polyprezinc Granules) is a mucosal protective agent with a unique mechanism of action. It promotes cell proliferation by up-regulating the expression of IGF-I mRNA to achieve effective treatment of ulcers and accelerate ulcer healing.

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