Application of Brazil green propolis in preparation of PDT antitumor synergistic drugs
A synergistic and anti-tumor technology, applied in the field of medicine, can solve the problems of weakening the curative effect, damaging tumor cells, and increasing the level.
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Embodiment 1
[0023] Experimental purpose: through the cytotoxicity test and the calculation of the synergy index to show that the BGP combined with PDT method has a synergistic effect on anti-tumor.
[0024] experimental design:
[0025] On the human skin squamous cell carcinoma cell line A431, the MTT method was used to determine the cytotoxicity of different doses of BGP, PpIX-PDT and BGP combined with PpIX-PDT. The results are shown in Table 1.
[0026]
[0027] Table 1
[0028] Result analysis: When BGP 62.5μg / ml was administered alone, the cell survival rate was 73.0±2.0%, and when the concentration reached 75μg / ml, the cell survival rate was only 62.1±2.2%. When PpIX 1.8μM, 2.7μM or 4.0μM was administered alone, the cell survival rate was 74.9±0.9%, 65.9±1.8% and 34.1±2.3%, respectively. When BGP was used in combination with PpIX-PDT, the survival percentage of A431 cells was significantly lower than that of BGP alone group and PpIX-PDT alone group.
[0029] To detect the synergistic effect...
Embodiment 2
[0034] Experimental purpose: To demonstrate that BGP combined with PDT has a synergistic effect on anti-tumor by measuring the expression levels of apoptosis and anti-apoptotic proteins and pro-apoptotic proteins.
[0035] Experimental design: qualitatively and quantitatively determined cell apoptosis caused by BGP, PpIX-PDT and BGP combined with PpIX-PDT on A431 cells by Annexin V-FITC / PI staining. The expression of anti-apoptotic protein Bcl-xL and pro-apoptotic protein Bax, NOXA, and cleaved caspase-3 were detected by Western Blot method.
[0036] (1) After A431 cells are treated with BGP (75μg / ml), PpIX (2.7μM) or BGP combined with PpIX-PDT, the ratio of live cells, early apoptotic cells, late apoptotic cells and necrotic cells. Each group received laser irradiation with an intensity of 27mW / cm2. The result is figure 1 with figure 2 Shown.
[0037] Result analysis: At the experimental dose, almost all groups did not cause necrosis of A431 cells, and apoptosis was the main form...
Embodiment 3
[0041] Experimental purpose: To demonstrate that BGP combined with PDT can reduce cell inflammation by measuring the protein expression level of NF-κB / COX-2 signaling pathway.
[0042] experimental design:
[0043] Western Blot method was used to detect the expression of BGP, PpIX-PDT and BGP combined with PpIX-PDT on the protein expression of NF-κB / COX-2 signaling pathway in A431 cells. .
[0044] Western Blot method was used to determine the changes in the expression levels of inflammation-related proteins such as p-IKKα / β, NF-κB, COX-2, etc. after A431 cells were treated with BGP (75μg / ml), PpIX (2.7μM) and BGP combined with PpIX-PDT . Use β-Actin as internal control. Each group received laser irradiation with an intensity of 27mW / cm2. The result is Figure 4 Shown.
[0045] Result analysis: When only PDT was given, p-IKKα / β in A431 cells increased significantly, the expression of NF-κB was up-regulated, and the protein level of its downstream effect target COX-2 also increased...
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