阅读说明：本技术 线粒体移植在治疗原发性扩心病中的应用 (Application of mitochondria transplantation in treating primary dilated cardiomyopathy ) 是由 葛均波 孙爱军 孙晓垒 于 2020-04-30 设计创作，主要内容包括：本发明属于原发性扩心病治疗领域,具体涉及线粒体移植在治疗原发性扩心病中的应用。本发明获得人源诱导多功能干细胞,体外分化诱导成跳动的心肌细胞,分离获得人源心肌细胞线粒体,同时分离小鼠心肌线粒体,分别获得人源和鼠源线粒体备用。本发明通过腹腔注射阿霉素构建小鼠扩心病模型,并在小鼠左心室心肌表层多点移植人源和鼠源线粒体,6天后超声心动图检测小鼠心脏功能,组织病理学检测小鼠心肌结构变化。研究结果显示,阿霉素诱导的小鼠扩心病模型构建过程中,移植人源或鼠源心肌细胞线粒体能够显著增强心肌收缩和舒张功能,抑制心脏双室心腔扩张。(The invention belongs to the field of primary dilated cardiomyopathy treatment, and particularly relates to application of mitochondrial transplantation in treatment of primary dilated cardiomyopathy. The invention obtains human-derived induced multifunctional stem cells, differentiates and induces the stem cells into beating myocardial cells in vitro, separates and obtains human-derived myocardial cell mitochondria, and simultaneously separates mouse myocardial mitochondria to respectively obtain human-derived mitochondria and mouse-derived mitochondria for later use. The invention constructs a mouse heart disease model by injecting adriamycin into abdominal cavity, transplants human and mouse mitochondria on the surface layer of the cardiac muscle of the left ventricle of the mouse at multiple points, detects the cardiac function of the mouse by echocardiogram after 6 days, and detects the structural change of the cardiac muscle of the mouse by histopathology. Research results show that in the process of constructing a mouse heart enlargement disease model induced by adriamycin, human or mouse myocardial cell mitochondria are transplanted to remarkably enhance the myocardial contraction and relaxation functions and inhibit the expansion of the heart double-chamber heart cavity.)

1. Application of mitochondria preparation in preparing medicine for treating heart failure.

2. The use according to claim 1, wherein the heart failure is caused by myocardial dysfunction.

3. Use according to claim 2, wherein the myocardial dysfunction is caused by doxorubicin injections.

4. Use according to claim 3, wherein the myocardial dysfunction is myocardial contraction and relaxation dysfunction.

5. Application of mitochondria preparation in preparing medicine for treating primary dilated cardiomyopathy.

6. The use according to any one of claims 1 to 4 or claim 5, wherein the mitochondria are autologous mitochondria.

7. The use of claim 6, wherein the mitochondrial preparation comprises physiological saline.

8. The use of claim 7, wherein the mitochondrial preparation is present at a concentration of 10 per 100 μ l⁵And (4) mitochondria.

Technical Field

The invention belongs to the field of primary dilated cardiomyopathy treatment, and particularly relates to application of mitochondrial transplantation in treatment of primary dilated cardiomyopathy.

Background

The Dilated Cardiomyopathy (DCM) is a primary myocardial disease and mainly shows congestive heart failure caused by ventricular cavity enlargement and contraction function reduction, the death rate of the dilated cardiomyopathy accounts for 20% within one year and 56% within four years according to statistics, the heart failure is delayed and the life of a patient is prolonged due to the complex pathogenic factors of the dilated cardiomyopathy, most of the cases have no clear treatment measures, the medicines such as ACE inhibitor, diuretic, β receptor blocker, digitalis, anticoagulant and the like are clinically used only by relieving symptoms, the incidence rate of the heart failure is still 40% within two years, heart transplantation is needed, mitochondrial dysfunction caused by genes or non-genes is the main cause of the contraction function abnormality of the left ventricle and is an important factor for the occurrence and development of the dilated cardiomyopathy, and the current treatment strategy of the dilated myocardium is immature due to the unclear pathogenic factors of the dilated cardiomyopathy.

The mitochondria transplantation is an intervention treatment mode, and particularly, normal mitochondria are obtained from other parts of a patient and then directly injected into abnormal tissue parts of the mitochondria, so that damaged mitochondria can be replaced, damaged cells can be cured, and the organ function can be recovered to be normal. The existing evidence indicates that the mitochondrial transplantation has completely exposed the horn in the treatment of various mitochondrial defects and diseases related to dysfunction, and the treatment means opens a new approach for neuroprotection. In addition, various pathological processes such as alzheimer's disease, parkinson's disease, muscular dystrophy, age-related neuropathy, traumatic neuropathy, and concussion are also associated with mitochondrial dysfunction. The application of mitochondrial transplantation in clinic will become more and more popular, and the unique treatment scheme conforms to the precise medical concept of 'treating self diseases with self substances'. However, there is no record of the treatment of primary dilated cardiomyopathy by mitochondrial transplantation.

Disclosure of Invention

The invention separates and obtains human-derived myocardial cell mitochondria, and simultaneously separates mouse myocardial mitochondria to respectively obtain human-derived mitochondria and mouse-derived mitochondria for later use. A mouse heart disease model is constructed by injecting adriamycin into the abdominal cavity, human-derived and mouse-derived mitochondria are transplanted on the surface layer of the cardiac muscle of the left ventricle of the mouse at multiple points, the heart function of the mouse is detected by an echocardiogram after 6 days, and the structural change of the cardiac muscle of the mouse is detected by histopathology.

In a first aspect, the invention provides the use of a mitochondrial preparation for the manufacture of a medicament for the treatment of dilated cardiomyopathy.

The effect of adopting above-mentioned technical scheme is: in the process of constructing a mouse heart enlargement disease model induced by adriamycin, the transplanted human or mouse cardiomyocyte mitochondria can obviously enhance the myocardial contraction and relaxation functions and inhibit the expansion of the heart cavities of the two chambers of the heart.

The method can be further improved on the basis of the scheme, and the specific improvement is as follows:

further, the mitochondria are autologous mitochondria. Transplantation with autologous mitochondria avoids rejection from allogeneic mitochondria.

Further, the mitochondrial preparation contains physiological saline.

Further, the heart failure is caused by myocardial dysfunction in dilated cardiomyopathy. The mitochondrial preparation provided by the invention can effectively improve myocardial dysfunction caused by dilated cardiomyopathy. At the same time provide

Further, the cardiomyopathy myocardial dysfunction is caused by adriamycin injection.

Further, the myocardial dysfunction refers to myocardial contraction and relaxation dysfunction. The mitochondrial preparation provided by the invention can obviously enhance the myocardial contraction and relaxation functions.

In a second aspect, the invention also provides a method of delivering a mitochondrial agent to a target of a tissue, in particular, administering a therapeutically effective dose of a mitochondrial agent to the target of the tissue.

The effect of adopting above-mentioned technical scheme is: energy-related contractile dysfunction caused by mitochondrial dysfunction can be effectively alleviated by exogenous supply of mitochondria.

The method can be further improved on the basis of the scheme, and the specific improvement is as follows:

further, the tissue is a heart; preferably, the tissue is the surface layer of the myocardium.

Further, the mitochondria are autologous mitochondria, and the mitochondrial preparation contains physiological saline.

Further, the mitochondrial preparation has a concentration of 10 per 100 microliters⁵And (4) mitochondria.

Compared with the prior art, the invention has the technical effects that:

(1) the invention focuses on the common point of the contraction dysfunction related to the energy supply of the myocardial cells of the dilated cardiomyopathy, realizes the mitochondrial transplantation, can effectively improve the heart dysfunction related to the dilated cardiomyopathy, ensures the energy supply of the myocardial function, obviously enhances the contraction and relaxation functions of the myocardial, inhibits the expansion of the heart double chambers and the heart cavities, inhibits the occurrence of heart failure and improves the life quality of the patient with the dilated cardiomyopathy.

(2) The invention separates and obtains the cardiac muscle cell mitochondria differentiated from the human-derived induced multifunctional stem cells, and the mitochondria contains 10 per 100 microliter⁵Mitochondrial suspension of mitochondria dissolved in normal saline is injected on the surface of the myocardial layer of the left ventricle, meanwhile, adriamycin is injected in the abdominal cavity according to 15mg/kg to construct a mouse heart enlargement disease model, and the transplantation verification of the mouse heart enlargement disease model is carried out, thereby effectively popularizing the clinical application of mitochondrial transplantation.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments will be briefly described below.

FIG. 1 technical roadmap of the present application

FIG. 2 evaluation of the therapeutic Effect of mouse cardiomyocyte-derived mitochondrial transplantation on dilated cardiomyopathy

The transplantation of mitochondria A obviously increases the left ventricular ejection fraction of the heart of a mouse with the heart dilated disease induced by adriamycin;

b, mitochondrial transplantation obviously increases the heart short axis shortening rate of the adriamycin-induced dilated cardiomyopathy mice;

c and D ventricular septum thickness during systole and diastole of mice in different treatment groups;

e and F mitochondria transplantation obviously reduces the heart contraction and diastole left ventricular internal diameter of mice with the heart dilatation disease induced by adriamycin;

g and H different treatment groups mice contracted and relaxed with left ventricular wall thickness.

CON control, DOX doxorubicin-treated, DOX + MITO doxorubicin-treated merged mitochondrial transplantation, S systolic, d diastolic, IVS ventricular septal thickness, L VID left ventricular internal diameter, L VPW left ventricular wall thickness p <0.05, p <0.01, p < 0.0001.

FIG. 3 transplantation of mitochondria of human cardiomyocytes for effectively improving cardiac function of mice with dilated cardiomyopathy

The transplantation of mitochondria A obviously increases the heart left ventricular ejection fraction of a mouse with Doudou dilated cardiomyopathy;

b, mitochondrial transplantation obviously increases the heart short axis shortening rate of the adriamycin-induced dilated cardiomyopathy mice;

c and D ventricular septum thickness during systole and diastole of mice in different treatment groups;

e and F different treatment groups mice systolic and diastolic left ventricular internal diameter;

g and H different treatment groups mice in the systolic and diastolic phase left chamber wall thickness CON: control group, DOX: doxorubicin treatment group, DOX + MITO: doxorubicin treatment combined with mitochondrial transplantation S: systolic phase, d: diastolic phase, IVS: chamber separation thickness, L VID: left chamber internal diameter, L VPW: left chamber wall thickness p <0.05, p <0.01, p < 0.0001.

Detailed Description

The invention is further described below with reference to the accompanying drawings. The invention will be better understood from the following examples. However, it is easily understood by those skilled in the art that the description of the embodiment is only for illustrating and explaining the present invention and is not for limiting the present invention described in detail in the claims.