CN104107419A

CN104107419A - Application of ghrelin in prevention or/and treatment of radiation-induced lung injury

Info

Publication number: CN104107419A
Application number: CN201410384594.0A
Authority: CN
Inventors: 卢铀; 薛建新; 周麟
Original assignee: West China Hospital of Sichuan University
Current assignee: West China Hospital of Sichuan University
Priority date: 2014-05-29
Filing date: 2014-08-06
Publication date: 2014-10-22

Abstract

The invention provides a novel medicinal application of ghrelin, namely application of ghrelin in prevention or/and treatment of radiation-induced lung injury, radiation-induced pneumonia and late pulmonary fibrosis. In-vivo experiments verify that the ghrelin is effective in the fields of prevention or/and treatment of radiation-induced lung injury, radiation-induced pneumonia and late pulmonary fibrosis. According to the novel medicinal application of the ghrelin, the ghrelin is creatively applied to prevention or/and treatment of radiation-induced lung injury, radiation-induced pneumonia and late pulmonary fibrosis, and a novel choice is provided to clinical treatment.

Description

Leptin prevention or/and the purposes in radiotherapy injury of lung

Technical field

The present invention relates to the new purposes of one of Leptin, be specifically related to Leptin prevention or/and the purposes in radiotherapy injury of lung.

Background technology

Pulmonary carcinoma is that current sickness rate is high, and therapeutic effect is poor, one of malignant tumor that cancer related mortality is the highest.Radiotherapy is the important means of lung cancer therapy, and approximately 50% patients with lung cancer need to be accepted radiotherapy.Induced lung injury (Radiation-induced lung injury, RILI) is the modal complication of radiotherapy in lung cancer, and incidence rate reaches 10%～20%.Induced lung injury Early manifestation is radiation pneumonia, and later stage progress is radiation fibrosis of lung, causes death when serious.At present, RILI still lacks effective means of prevention, adrenocortical hormone, broad ectrum antibiotic, bronchodilator and immunosuppressant are still the Main Means for the treatment of acute radiation pneumonitis, and radiation fibrosis of lung to late period, so far without effectively preventing and treatment measure.Therefore, find the prevention of a kind of effective measures or alleviate RILI significant for the effect that improves radiotherapy of lung cancer.

Leptin (Ghrelin), is made up of 28 aminoacid, is the specific endogenic ligand of secretagogue receptor (Growth hormone secretagogue receptor, GHSR).Leptin decapacitation effectively promotes, outside growth hormone secretion, also to participate in adjusting energy metabolism, gastrointestinal function and cardiovascular function etc.In recent years, have the bibliographical information of Leptin in other function aspects, for example, the Leptin of restructuring has been proved in clinical trial can relieving anorexia disease, stomach spasm, cachexia and heart failure.In addition, Leptin has antiinflammatory and anti-fibrosis effect, studies have reported that, external source give Leptin can alleviate the acute lung injury that bleomycin or septicemia cause ( nakazato M.etal. eur J? pharmacol.2011,672 (1-3): 153-8; Wang P.et al.Am J Respir Crit Care Med2007,176:805-13), but do not report Leptin for preventing or/and radiotherapy injury of lung at present.Therefore, the new medical usage of exploitation Leptin, is applied in Leptin prevention in a creative way or/and in radiotherapy injury of lung, for clinical treatment induced lung injury provides a kind of new selection.

Summary of the invention

In order to solve the problem of prior art, the invention provides a kind of new medical usage of Leptin, Leptin in prevention or/and the purposes in radiotherapy injury of lung, radiation pneumonia, pulmonary fibrosis in late period.

The invention provides Leptin prevents or/and the purposes in the medicine of radiotherapy injury of lung in preparation.

The invention provides Leptin prevents or/and the purposes in the medicine for the treatment of radiation pneumonia in preparation.

The invention provides Leptin prevents or/and the purposes in the medicine for the treatment of pulmonary fibrosis in late period in preparation.

Leptin is made to a kind of polypeptide drugs, is to be active component by Leptin, the preparation that adds pharmaceutically acceptable adjuvant or complementary composition to be prepared from.

Described preparation is oral formulations or ejection preparation.

Described oral formulations is tablet or capsule.

Described ejection preparation is injection or freeze-dried powder.

The present invention has significant preventive and therapeutic action through a large amount of Leptins that experiment showed, to induced lung injury, radiation pneumonia, pulmonary fibrosis in late period, for clinical treatment provides a kind of new peptide drug.

Brief description of the drawings

January after the full lung of the foundation of Fig. 1 induced lung injury mouse model: A irradiates; B matched group (× 200).

Fig. 2 C57 mouse lung is organized H & E dyeing (× 200).

Fig. 3 Leptin is on the apoptotic impact of lung tissue.A lung tissue TUNEL dye (× 680); B lung tissue CLEAVED CASPASED-3 WESTERN BLOT detects.

The impact of Fig. 4 Leptin on lung tissue DNA Damage.A lung tissue cell Γ H2AX immunofluorescence dyeing (× 680); B lung tissue Γ H2AX WESTERN BLOT detects.

The impact that Fig. 5 Leptin produces lung tissue ROS.

The impact of Fig. 6 Leptin on blood plasma inflammatory factor.

The therapeutic effect of Fig. 7 H & E staining examine Leptin to radiation fibrosis of lung.

Fig. 8 MASSON ' S trichrome stain detects the impact of Leptin on radiation fibrosis of lung.

Fig. 9 liquid-phase chip detects the impact of Leptin on fibrosis factor TGF-β 1.

Detailed description of the invention

In following embodiment, Leptin used is synthesized by biotech firm, can directly buy commercially available product.

Embodiment 1

Leptin is dissolved with PBS, be mixed with injection.

Effect taking this injection as example research Leptin to prevention and radiotherapy injury of lung.

The zoopery of experimental example 1 medicine of the present invention

One, experimental technique

1. male mice gives full lung 16Gy irradiation after conforming;

2. male mice is divided into 4 groups at random, every group of n only;

Normal group (PBS): 0.65mg/kg PBS, i.p., the next day scheme;

Simple Leptin group (G): 0.65mg/kg Leptin, i.p., the next day scheme;

Irradiation+buffer group (RT+PBS): 0.65mg/kg PBS, i.p., the next day scheme; Injected for the first time before 24 hours and give chest irradiation 16Gy1 time;

Irradiation+Leptin treatment group (RT+G): 0.65mg/kg Leptin, i.p., the next day scheme; Injected for the first time before 24 hours and give chest irradiation 16Gy 1 time;

Every group of mice n >=5.

Two, experiment (induced lung injury mouse model) in body

1. the foundation of induced lung injury model mice

After 16Gy Irradiation C57 mice January, there is obvious injury of lung performance in mouse lung tissue, and pulmonary edema, hyperemia, alveolar septum broadening also have massive inflammatory cells infiltrated.Prove that mice induced lung injury model is successfully established, and all takes this dosage modeling later under 16Gy Irradiation.The results detailed in Fig. 1.

The impact of 2.H & E staining examine Leptin on early stage induced lung injury degree

C57 mice pre-irradiation is injected Leptin or PBS in 1 day, and administration next day of afterwards, put to death mice after 30 days, adopts H & E colouring method to detect the impact of Leptin on the early stage induced lung injury of mice.Result shows that Leptin can significantly alleviate the induced lung injury of mice.The results detailed in Fig. 2.

3. Leptin is on the apoptotic impact of lung tissue

After Leptin treatment, within the 30th day, collect the tissue specimen of experimental group and control group mice lung, adopt Tunnel dyeing and Western Blot to detect Leptin to the apoptotic impact of lung tissue.Testing result shows: more independent 16Gy combination radiotherapy group, Leptin can obviously suppress the expression of Cleaved caspase3, is illustrated as Leptin and can suppresses lung tissue apoptosis.The results detailed in Fig. 3.

4. the impact of Leptin on lung tissue DNA Damage

Adopt immunofluorescence dyeing to detect the formation of lung tissue cell γ H2AX focus, adopt Western Blot to detect the expression of lung tissue γ H2AX albumen.Testing result shows: more independent 16Gy combination radiotherapy group, Leptin can suppress the DNA damage of lung tissue cell.The results detailed in Fig. 4.

5. the impact that Leptin produces lung tissue ROS

Adopt active oxygen detection kit to detect each processed group lung tissue ROS level.Testing result shows: more independent combination radiotherapy group, Leptin can suppress the generation of the ROS of lung tissue.The results detailed in Fig. 5.

6. the impact of Leptin on blood plasma inflammatory factor

Blood plasma inflammatory factor plays an important role in the evolution of RILI, and when after RADI lung tissue, the secretion of inflammatory factor increases, also corresponding rising of the inflammatory factor level in peripheral blood.Studies show that blood plasma inflammatory factor level can hinder degree by indirect reaction damaged lung tissue.We adopt liquid-phase chip technology to detect the concentration of irradiating inflammatory factor in latter 30 days each experimental mice blood plasma, comprising: TGF-β 1, TGF-β 2, GM-CSF, IFN γ, IL-6, IL-1 α, IL-β, IL-2, TNF-α.Result shows that Leptin pretreated group blood plasma inflammatory factor is all lower than simple Irradiation group, wherein TGF-β 1, TGF-β 2, IFN γ, IL-1 α, IL-β, IL-2, the concentration of TNF-α all has significant difference p<0.05, and the concentration of GM-CSF and IL-6 has no obvious significant difference.The results detailed in Fig. 6.

7. the impact of Leptin on the pulmonary fibrosis of induced lung injury model mice

(1) therapeutic effect of H & E staining examine Leptin to radiation fibrosis of lung

As shown in Figure 7: after 16Gy roentgenization 6 months, i.e. Leptin treatment finishes latter 5 months, and obviously damaging appears in mouse lung tissue, alveolar septum obvious broadening during compared with 1 month, inside there is massive inflammatory cells infiltrated, alveolar form destruction, abnormal, between lung, hypertrophy is obvious.In Leptin treatment group, damage more simple Irradiation group and alleviate to some extent.

(2) Masson ' s trichrome stain detects the impact of Leptin on radiation fibrosis of lung

Radiation pneumonia and radiation fibrosis of lung are two major parts of RILI, and generally, radiation pneumonia occurs in after roentgenization 2-4 month, after radiation fibrosis of lung occurs in and irradiates 4-6 month.In irradiation latter 6 months, we put to death mice and detect the degree of its lung fibrosis.Masson ' s dyeing is conventional collagen fiber colouring method, and in the method, collagen fiber are dyed for blueness, and nuclei dyeing is blue brown, and redness has represented erythrocyte and muscle fiber.As shown in Figure 8: in simple Irradiation group (RT+PBS), alveolar septum and bronchus be visible a large amount of collagen deposition around, alleviates to some extent at Leptin treatment group collagen deposition.

(3) liquid-phase chip detects the impact of Leptin on fibrosis factor TGF-β 1

In the pathophysiological process of RILI, plasma fibronectin factor TGF-β 1 plays a part very important.Peripheral blood T GF β 1 level has indirect predictions and assesses damaged lung tissue the effect of hindering degree.We have detected the concentration of 6 months the central TGF-β 1 of experimental mice blood plasma after Irradiation.Result shows: the level of Leptin processed group mice plasma TGF-β 1 is starkly lower than simple Irradiation group, and difference has statistical significance (p<0.05), the results detailed in Fig. 9.

Claims

1. Leptin prevents or/and the purposes in the medicine of radiotherapy injury of lung in preparation.

2. Leptin prevents or/and the purposes in the medicine for the treatment of radiation pneumonia in preparation.

3. Leptin prevents or/and the purposes in the medicine for the treatment of pulmonary fibrosis in late period in preparation.

4. according to the purposes described in claim 1-3 any one, it is characterized in that: described medicine is to be active component by Leptin, the preparation that adds pharmaceutically acceptable adjuvant or complementary composition to be prepared from.

5. purposes according to claim 4, is characterized in that: described preparation is oral formulations or ejection preparation.

6. purposes according to claim 5, is characterized in that: described oral formulations is tablet or capsule.

7. purposes according to claim 5, is characterized in that: described ejection preparation is injection or freeze-dried powder.