阅读说明：本技术 灵菌红素在制备***肌瘤病细胞增殖抑制剂中的应用 (Application of prodigiosin in preparation of cell proliferation inhibitor for lymphangiomatosis ) 是由 赵银娟 薛斌 程琪 沈紫竹 于 2019-11-29 设计创作，主要内容包括：本发明公开了灵菌红素在制备淋巴管肌瘤病细胞增殖抑制剂中的应用,涉及生物医药领域。本发明通过生物工程技术制得灵菌红素,并使用显微镜观察细胞形态变化,cck-8法测定细胞活性变化,观察灵菌红素对淋巴管肌瘤病细胞Tsc2null增殖的抑制作用,发现,24h之内,灵菌红素对Tsc2null的抑制效果明显高于Tsc2wt,2h之内,Tsc2null的细胞活性均和灵菌红素的浓度呈反比,灵菌红素浓度越高,细胞活性越低,灵菌红素对淋巴管肌瘤病细胞的增殖具有显著的抑制作用,可见,灵菌红素在制备淋巴管肌瘤病细胞的增殖抑制剂或抗淋巴管肌瘤病细胞的增殖药物中即将具有广泛的应用。(The invention discloses application of prodigiosin in preparation of a lymphangiomyomatosis cell proliferation inhibitor, and relates to the field of biological medicines. The method prepares the prodigiosin through a biological engineering technology, observes the change of cell morphology by using a microscope, measures the change of cell activity by a cck-8 method, observes the inhibition effect of the prodigiosin on the proliferation of a lymphangiosarcomatosis cell Tsc2null, and discovers that within 24h, the inhibition effect of the prodigiosin on the Tsc2null is obviously higher than that within Tsc2wt and 2h, the cell activity of the Tsc2null is in inverse proportion to the concentration of the prodigiosin, the higher the concentration of the prodigiosin is, the lower the cell activity is, the prodigiosin has obvious inhibition effect on the proliferation of the lymphangiomatosis cell, and the prodigiosin can be widely applied to the preparation of a proliferation inhibitor of the lymphangiomatosis cell or a proliferation medicine for resisting the lymphangiomatosis cell.)

1. Application of prodigiosin in preparing cell proliferation inhibitor for lymphangiomatosis is provided.

2. The use of claim 1, wherein the lymphangiosarcomatosis cells are Tsc2null cells.

3. The use as claimed in claim 1, wherein the concentration of the rubicin in the inhibitor is 0.01-100 μmol/L.

4. Application of prodigiosin in preparing medicines for inhibiting cell proliferation of lymphangiomatosis is provided.

5. The use according to any one of claims 1 to 4, characterized in that the prodigiosin is prepared by a process comprising: centrifuging the serratia marcescens culture solution, and separating supernatant and precipitate; leaching the lower layer precipitate with methanol, and centrifuging to obtain haematochrome; rotary evaporating with rotary evaporator to obtain dark purplish red pigment methanol solution, and processing with evaporator to obtain filtrate C₁₈And (3) performing reverse phase semi-preparative HPLC (high performance liquid chromatography) repeated purification, dissolving the pigment in absolute ethyl alcohol, and performing vacuum low-temperature high-speed freeze-drying to obtain the pure prodigiosin.

Technical Field

The invention relates to the field of biological medicines, in particular to application of prodigiosin in preparation of a cell proliferation inhibitor for lymphangiomyomatosis.

Background

Lymphangiomyomatosis (LAM) is a rare disease mainly caused by diffuse cystic lung disease, mainly occurs in women in the reproductive age, and the pathology is mainly characterized by abnormal proliferation and infiltration of smooth muscle-like cells. LAM has various clinical manifestations, and can affect kidney, lymphatic vessels, lymph nodes, spleen, liver, etc. besides characteristic cystic changes in the lung. Studies have shown that LAM is derived from mutations in the TSC1/TSC2 gene in patients, with TSC2 being the more common. TSC is a tumor suppressor gene, a TSC-LAM patient has mutation of a TSC gene at one site in nature, and after birth, the TSC gene at the second site is obtained again to cause disease. After the 2 allelic TSC (particularly TSC2) genes are completely inactivated, the inhibition is lost, and then the downstream mTOR pathway is abnormally activated, so that the LAM cell is stimulated to abnormally grow, proliferate, migrate and metastasize, and the structural damage and the dysfunction of the organ systems in the lung and outside the lung are caused.

Prodigiosin (Prodigiosin) is a red antibiotic found in various bacteria and actinomycetes, and the skeleton of the Prodigiosin is characterized by containing a large conjugated system of a three-pyrrole ring, so that the Prodigiosin has various activities such as cancer resistance, fungus resistance, bacteria resistance, protozoa resistance, immunosuppression and the like. The antitumor activity of prodigiosin has been of particular interest in recent years, and the average IC of prodigiosin has been found by the national cancer institute (NSI) Melvin et al₅₀At 2.1. mu. mol/L, it was found to be resistant to 57 different human cancer cells and induce apoptosis. At present, the inhibitory effect of prodigiosin on the proliferation of LAM disease cells Tsc2null has not been studied.

Disclosure of Invention

Aiming at the technical defects, the invention aims to provide the application of prodigiosin in preparing a cell proliferation inhibitor for lymphangiomyopathy, and the prodigiosin has obvious inhibition effect on the proliferation of LAM disease cells Tsc2 null.

In order to achieve the purpose of the invention, the invention adopts the technical scheme that:

application of prodigiosin in preparing cell proliferation inhibitor for lymphangiomatosis is provided.

Further, the lymphangiosarcomatosis cell is a Tsc2null cell.

Further, the concentration of the rubicin in the inhibitor is 0.01-100 mu mol/L.

Application of prodigiosin in preparing medicines for inhibiting cell proliferation of lymphangiomatosis is provided.

Further, the preparation method of the prodigiosin comprises the following steps: centrifuging the serratia marcescens culture solution, and separating supernatant and precipitate; leaching the lower layer precipitate with methanol, and centrifuging to obtain haematochrome; rotary evaporating with rotary evaporator to obtain dark purplish red pigment methanol solution, and processing with evaporator to obtain filtrate C₁₈And (4) performing reversed-phase semi-preparative HPLC (high performance liquid chromatography) repeated purification, dissolving the finally obtained pigment in absolute ethyl alcohol, and performing vacuum low-temperature high-speed freeze-drying.

Has the advantages that: the invention prepares the prodigiosin by a biological engineering technology, observes the change of cell morphology by using a microscope, measures the change of cell activity by a cck-8 method, observes the inhibition effect of the prodigiosin on the proliferation of a LAM disease cell Tsc2null, and discovers: within 24h, the inhibition effect of prodigiosin on Tsc2null is obviously higher than that of Tsc2wt, within 2h, the cell activity of Tsc2null is in inverse proportion to the concentration of prodigiosin, the higher the concentration of prodigiosin, the lower the cell activity, the obvious inhibition effect on the proliferation of lymphatic vessel sarcoidosis cells is achieved, and the prodigiosin can be widely applied to the preparation of proliferation inhibitors of lymphatic vessel sarcoidosis cells or proliferation drugs for resisting the lymphatic vessel sarcoidosis cells.

Drawings

FIG. 1 is a graph showing the inhibitory effect of prodigiosin at various concentrations for 24h on Tsc2null cells;

FIG. 2 is a graph showing the inhibitory effect of prodigiosin at various concentrations of 48h on Tsc2null cells;

FIG. 3 is a graph showing the inhibitory effect of prodigiosin at various concentrations for 72h on Tsc2null cells;

FIG. 4 is a graph showing the inhibitory effect of prodigiosin at various concentrations for 24h on Tsc2wt cells;

FIG. 5 is a graph showing the inhibitory effect of prodigiosin at various concentrations for 48h on Tsc2wt cells;

FIG. 6 is a graph showing the inhibitory effect of prodigiosin at various concentrations for 72h on Tsc2wt cells;

FIG. 7 is a graph showing absorbance values of Tsc2null cells measured by the CCK-8 method after various treatments;

FIG. 8 is a graph showing the absorption values of Tsc2wt cells measured by the CCK-8 method after various treatments.

Detailed Description

The present invention is further illustrated by the following specific examples, which are not intended to be limiting.