BMPR (bone morphogenetic protein receptor) nano-probe for targeting hepatic fibrosis as well as preparation method and application of BMPR nano-probe

Abstract

The invention discloses a BMPR nano probe for targeting hepatic fibrosis as well as a preparation method and application of the BMPR nano probe, and belongs to the technical field of hepatic fibrosis targeting drugs. The BMPR nanoprobe prepared by the invention has good hepatic stellate cell targeting ability, and the targeting effect is enhanced along with increase of the concentration of the BMPR nanoprobe and prolonging of incubation time; the higher the severity of hepatic fibrosis is, the more obvious the targeting effect is; the relaxation performance is good; the compound has obvious cytotoxicity to hepatic stellate cells, has relatively strong nuclear magnetic resonance imaging capability, and can be used for monitoring hepatic fibrosis and developing hepatic fibrosis treatment medicaments; the compound can effectively inhibit the expression of each protein in an HIF-1alpha / TGF-beta1 / Smad pathway, further inhibits the activation of hepatic stellate cells, effectively improves the hypoxia microenvironment of hepatic fibers, and has an excellent treatment effect on hepatic fibrosis; the provided preparation method is simple and convenient and has strong operability; and the synthesized product is high in stability and good in repeatability.

Description

Technical Field

[0001] The present invention relates to the technical field of liver fibrosis targeting drugs, and particularly to a BMPR nanoprobe targeting liver fibrosis, a preparation method thereof, and an application thereof. Background Art

[0002] Liver fibrosis is a common chronic liver disease, mainly caused by two types of chronic liver injuries: hepatotoxic injury and cholestatic injury. Early liver fibrosis is a reversible process. If not treated in time, it will develop into liver cirrhosis, liver failure, and even liver cancer. At this time, the liver structure and function will suffer irreversible damage, even threatening the life of the patient.

[0003] At present, the diagnostic methods for liver fibrosis include ultrasound elastography (UE) and magnetic resonance elastography (MRE), but both have limitations: the accuracy is deviated, and image processing requires additional software equipment. Traditional treatment methods have weak specificity, poor targeting, large side effects, easy drug resistance, and insignificant curative effects. The existing diagnostic and treatment methods are in an unsatisfied state for liver fibrosis. However, during the research process, it was found that the severity of liver fibrosis is positively correlated with the level of integrin αγβ3 receptor on the surface of hepatic stellate cells (HSCs). Integrin αγβ3 specifically recognizes the arginine-glycine-aspartic acid (RGD) tripeptide sequence.

[0004] After liver injury, it will lead to the activation of HSCs. Among them, the proliferation and transformation of activated HSCs are the core of liver fibrosis. Collagen-I, as the main component of the extracellular matrix (ECM) of liver fibrosis, is deposited in large amounts during the fibrosis process. The excessive deposition of ECM reduces the liver blood supply, creates a hypoxic microenvironment, promotes the activation of hematopoietic stem cells, and upregulates hypoxia-inducible factor-1α (HIF-1α). Subsequently, HSCs release various inflammatory factors such as TGF-β1, further enhancing the activation of HSCs, producing excessive α-SMA and collagen-I. At the same time, TGF-β1 upregulates integrin expression and enhances the adhesion of HSCs. Therefore, relieving liver fibrosis hypoxia can inhibit HIF-1α expression, block the TGF-β1 / Smad signaling pathway, and reduce ECM deposition. By downregulating the expression of HSCs HIF-1α, blocking the HIF-1α / TGF-β1 / Smad pathway, and inhibiting HIF-1α activation to treat liver fibrosis. Summary of the Invention

[0005] The purpose of the present invention is to provide a BMPR nanoprobe targeting liver fibrosis, a preparation method thereof, and an application thereof, so as to solve the problems of low accuracy, weak specificity, poor targeting, large side effects, and insignificant curative effects of current liver fibrosis diagnostic and / or treatment preparations.

[0006] To achieve the above object, the present invention provides a BMPR nanoprobe targeting liver fibrosis, which is prepared from bovine serum albumin, manganese pentacarbonyl bromide, dopamine, and arginine-glycine-aspartic acid tripeptide.

[0007] Preferably, the dopamine is maleimide polyethylene glycol dopamine and dopamine hydrochloride.

[0008] A method for preparing the BMPR nanoprobe targeting liver fibrosis as described above, the steps are as follows:

[0009] S1. Dissolve bovine serum albumin in deionized water, after mixing, add manganese pentacarbonyl bromide and stir overnight, centrifuge to discard the precipitate to obtain BSA@Mn(CO) 5 Br solution;

[0010] S2. Add arginine-glycine-aspartic acid tripeptide and maleimide polyethylene glycol dopamine to deionized water, stir in the dark on an ice bath to obtain a mixed solution;

[0011] S3. Dissolve the mixed solution obtained in step S2 and dopamine hydrochloride in Tris HCl, after mixing, add it to the BSA@Mn(CO) prepared in step S1 5 Br solution, stir in the dark for 12 h, centrifuge to discard the precipitate, and perform dialysis and ultrafiltration.

[0012] Preferably, in step S1, the mass-volume ratio of bovine serum albumin: manganese pentacarbonyl bromide: deionized water is 40 mg: 1 mg: 4 mL; the centrifugation conditions are 3000 rpm and centrifugation for 5 min.

[0013] Preferably, in step S2, the mass-volume ratio of arginine-glycine-aspartic acid tripeptide: maleimide polyethylene glycol dopamine: deionized water is 1 mg: 4 mg: 0.5 mL; the stirring conditions are 400 rpm and 30 min.

[0014] Preferably, the mass-volume ratio of maleimide polyethylene glycol dopamine: dopamine hydrochloride: Tris HCl is 3 mg: 12 mg: 2 mL; the pH of Tris HCl is 8.

[0015] Preferably, in step S3, the centrifugation conditions are 3000 rpm and centrifugation for 5 min; the dialysis uses deionized water as the dialysis solution, and the dialysis bag specifications are 44 mm and 14 kDa.

[0016] An application of the BMPR nanoprobe targeting liver fibrosis as described above in the preparation of an integrated diagnosis and treatment preparation.

[0017] An application of the BMPR nanoprobe targeting liver fibrosis as described above in the preparation of a liver fibrosis diagnostic preparation.

[0018] Use of a BMPR nanoprobe targeting liver fibrosis as described above in the preparation of a therapeutic agent for liver fibrosis.

[0019] Therefore, a BMPR nanoprobe targeting liver fibrosis provided by the present invention, its preparation method and application have the following specific technical effects:

[0020] (1) The BMPR nanoprobe prepared by the present invention has good targeting ability to hepatic stellate cells, and the targeting effect is enhanced with the increase of the concentration of the BMPR nanoprobe and the prolongation of the incubation time; moreover, the higher the severity of liver fibrosis, the more obvious the targeting effect; it has good relaxation performance; it has obvious cytotoxicity to hepatic stellate cells and strong nuclear magnetic resonance imaging ability, and can be used for monitoring liver fibrosis and the development of therapeutic agents for liver fibrosis;

[0021] (2) The BMPR nanoprobe prepared by the present invention can effectively inhibit the expression of each protein in the HIF-1α / TGF-β1 / Smad pathway, thereby inhibiting the activation of hepatic stellate cells, effectively improving the hypoxic microenvironment of liver fibrosis, and having excellent therapeutic effect on liver fibrosis;

[0022] (3) The preparation method provided by the present invention is simple and highly operable; the synthesized product has high stability and good repeatability. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following will briefly introduce the drawings required for the description of the embodiments of the present invention. Obviously, the following drawings are only some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0024] Figure 1 It is the transmission electron microscope picture of the BMP nanomaterial in Test Example 1 of the present invention;

[0025] Figure 2 It is the r1 relaxation map of BMPR in Test Example 2 of the present invention;

[0026] Figure 3 It is the confocal microscope photograph in Test Example 3 of the present invention;

[0027] Figure 4 It is the detection and analysis result in Test Example 4 of the present invention, wherein part a is the result of protein immunoblotting detection; part b is the quantification map of CollgenⅠ protein expression; part c is the quantification map of HIF-1α protein expression; part d is the quantification map of Smad4 protein expression; part e is the quantification map of α-SMA protein expression; part f is the quantification map of TGF-β protein expression;

[0028] Figure 5 It is the killing effect diagram of BMPR nanomaterials with different concentrations on hepatic stellate cells (LX-2) in Test Example 5 of the present invention;

[0029] Figure 6 It is the 9.4T magnetic resonance imaging effect after tail vein injection of BMPR nanomaterials into normal mice and liver fibrosis mice in Test Example 6 of the present invention: where part a is the magnetic resonance image of the mice; part b is the signal-to-noise ratio extreme value quantization diagram of normal mice; part c is the signal-to-noise ratio extreme value quantization diagram of liver fibrosis mice. Detailed implementation manners

[0030] The technical solution of the present invention will be further described below through the accompanying drawings and embodiments.

[0031] In order to make the purpose, technical solution and advantages of the present application clearer, more thorough and complete, the technical solution of the present invention will be clearly and completely described below through the accompanying drawings and embodiments. The following detailed descriptions are all descriptions of embodiments, aiming to provide further detailed descriptions of the present invention. Unless otherwise specified, all technical terms adopted by the present invention have the same meanings as those generally understood by those of ordinary skill in the art to which this application belongs.

[0032] The purpose of the present invention is to provide a construction method of a novel magnetic resonance nanoprobe targeting liver fibrosis. The present invention uses bovine serum albumin (BSA), manganese pentacarbonyl bromide (Mn(CO) 5 Br), dopamine hydrochloride (PDA), Mal-PEG-Do, and RGD as raw materials, and obtains (BSA@Mn(CO) 5 Br@PDA-RGD, BMPR) nanomaterials through a simple stirring reaction. The nanomaterial synthesis method provided by the present invention is simple, highly operable, has good nuclear magnetic imaging ability, and can monitor the severity of liver fibrosis through the MRI imaging effect and relieve the intracellular hypoxic environment to realize the integration of liver fibrosis diagnosis and treatment.

[0033] In order to enable those skilled in the art to more clearly understand the technical solution of the present application, the technical solution of the present application will be described in detail below in combination with specific embodiments.

[0034] The test materials used in the embodiments of the present invention are all conventional test materials in the art and can be obtained through commercial channels. The bovine serum albumin used in the present invention was purchased from Sinopharm Reagent Co., Ltd., CAS: 9048-46-8; dopamine hydrochloride was purchased from Macklin Co., Ltd., CAS: 62-31-7; manganese pentacarbonyl bromide was purchased from Bide Co., Ltd., CAS: 14516-54-2; human hepatic stellate cells (LX-2) were purchased from the National Experimental Cell Resource Sharing Platform; the mice used were purchased from the Experimental Animal Platform of the Medical Science and Technology Innovation Center of Shandong First Medical University, following the general rules of experimental animal welfare.

[0035] Example 1

[0036] Prepare a BMPR nanoprobe, and the specific steps are as follows:

[0037] (1) Prepare BSA@Mn(CO) 5 Br nanomaterials.

[0038] Dissolve 40 mg of bovine serum albumin (BSA) in 4 mL of deionized water and mix well by ultrasound, then add 1 mg of manganese pentacarbonyl bromide (Mn(CO) 5 Br), stir at 400 rpm overnight. After stirring, centrifuge at 3000 rpm for 5 min, discard the precipitate to obtain 4 mL of BSA@Mn(CO) 5 Br. Use the BCA quantification method to determine the BSA concentration in the supernatant. The result is 7.5 mg / mL, and the total amount of BSA in the supernatant is calculated to be 30 mg.

[0039] (2) Prepare BSA@Mn(CO) 5 Br@PDA-RGD nanomaterials (BMPR).

[0040] Add 1.5 mg of arginine-glycine-aspartic acid tripeptide (RGD) and 6 mg of maleimide polyethylene glycol dopamine (Mal-PEG-Do) (mass ratio 1:4) to 750 μL of deionized water, stir at 400 rpm in the dark on ice bath for 30 min, then dissolve the mixed solution of RGD and Mal-PEG-Do and 24 mg of dopamine hydrochloride (PDA) in 4 mL of Tris HCl solution with pH 8, mix well and add it to the BSA@Mn(CO) 5 Br aqueous solution (the mass ratio of PDA to BSA is 1:1), stir at room temperature in the dark for 12 h. After stirring, centrifuge at 3000 rpm for 5 min and discard the precipitate. Use deionized water as the dialysis solution, dialyze with a dialysis bag with a specification of 44 mm and 14 kDa for 24 h, and finally ultrafilter with a 15 mL, 10000 MWCO ultrafiltration tube to obtain 400 μL of solution BSA@Mn(CO) 5 Br@PDA-RGD nanomaterials (BMPR nanomaterials), Mn 2+ The concentration is 313 μg / mL.

[0041] Example 2

[0042] Prepare BSA@Mn(CO) 5 Br@PDA nanomaterials (BMP), and the specific steps are as follows:

[0043] (1) Prepare BSA@Mn(CO) 5Br nanomaterials.

[0044] Dissolve 40 mg of bovine serum albumin (BSA) in 4 mL of deionized water and mix well by ultrasonic treatment. Then add 1 mg of manganese pentacarbonyl bromide (Mn(CO) 5 Br), and stir at 400 rpm overnight. After stirring, centrifuge at 3000 rpm for 5 min, discard the precipitate, and obtain 4 mL of BSA@Mn(CO) 5 Br. Use the BCA quantification method to determine the BSA concentration in the supernatant. The result is 7.5 mg / mL, and the total amount of BSA in the supernatant is calculated to be 30 mg.

[0045] (2) Preparation of BMP nanomaterials.

[0046] Weigh 24 mg of PDA and 6 mg of Mal-PEG-Do and add them to 4 mL of Tris HCl solution with a pH of 8. Then add the 4 mL of BSA@Mn(CO) 5 Br solution obtained in step (1) (the mass ratio of PDA to BSA is 1:1), and stir in the dark for 12 h. After stirring, centrifuge at 3000 rpm for 5 min and discard the precipitate. Using deionized water as the dialysis solution, dialyze with a dialysis bag of 44 mm and 14 kDa specifications for 24 h. Finally, ultrafilter with a 15 mL, 10000 MWCO ultrafiltration tube to obtain 400 μL of solution, BMP nanomaterials, with a Mn 2+ concentration of 313 μg / mL.

[0047] Experimental Example 1

[0048] Use a transmission electron microscope to observe the BMP nanomaterials prepared in Example 2. The specific steps are as follows:

[0049] Take 20 μL of the supernatant containing BMP prepared in Example 2, add 1 mL deionized water, and use a pipette to add 40 μL dropwise onto a copper grid, dry it to prepare a transmission electron microscope detection sample, and then observe it with a transmission electron microscope.

[0050] The results are as Figure 1 shown. The size of the prepared BMP nanomaterials is about 30 nm.

[0051] Experimental Example 2

[0052] Detect the relaxation performance of the BMPR nanomaterials prepared in Example 1, as follows:

[0053] Absorb 10 μL of the BMPR prepared in Example 1, add 190 μL of deionized water to prepare a dispersion with a concentration of 0.3 mM, and then serially dilute it to BMPR nanomaterial dispersions with concentrations of 0.15 mM, 0.075 mM, 0.0375 mM, and 0.01875 mM. Measure the relaxation performance using a GY-PNMR-10 pulsed nuclear magnetic resonance imaging experimental instrument.

[0054] The results are as Figure 2 shown, indicating that the prepared BMPR nanomaterial has good relaxation performance and can be used for nuclear magnetic imaging.

[0055] Test Example 3

[0056] Investigate the uptake of BMPR prepared in Example 1 and BMP prepared in Example 2 by cells. The specific steps are as follows:

[0057] Seed human hepatic stellate cells (LX-2) in a confocal culture dish and treat the cells in five groups: (1) Control group, without adding any material; (2) BMPR 5 μg / mL (4 h) group, add 16 μL of the BMPR prepared in Example 1 to 984 μL of DMEM medium to make the concentration of Mn 5 μg / mL, mix well, and incubate in an incubator at 37 °C and 5% CO 2 for 4 h; (3) BMPR-2 group, add 32 μL of the BMPR prepared in Example 1 to 968 μL of DMEM medium to make the concentration of Mn 10 μg / mL, mix well, and incubate in an incubator at 37 °C and 5% CO 2 for 2 h; (4) BMPR-3 group, add 32 μL of the BMPR prepared in Example 1 to 968 μL of DMEM medium to make the concentration of Mn 10 μg / mL, mix well, and incubate in an incubator at 37 °C and 5% CO 2 for 4 h; (5) BMP group, add 32 μL of the BMP prepared in Example 2 to 968 μL of DMEM medium to make the concentration of Mn 10 μg / mL, mix well, and incubate in an incubator at 37 °C and 5% CO 2 for 4 h. After incubation, stain the cell nuclei with Hoechst dilution solution prepared according to the instructions for 20 min, and finally take pictures under a confocal microscope.

[0058] The results are as Figure 3 shown, indicating that the higher the concentration of the BMPR nanomaterial and the longer the incubation time, the more the material is taken up by the cells. And compared with BMP, under the same incubation time and the same treatment concentration conditions, the targeted material BMPR is taken up more by the cells.

[0059] Test Example 4

[0060] Examine the effects of the BMPR prepared in Example 1 and the BMP prepared in Example 2 on the expression of each protein in the TGF-β1 / Smad pathway. Using the Western Blot experiment, the TGF-β1 / Smad pathway proteins examined include CollgenⅠ (Col-I), HIF-1α, Smad4, TGF-β1, α-SMA, with GAPDH as the internal reference protein. The specific steps are as follows:

[0061] Seed human hepatic stellate cells (LX-2) in a 60 mm culture dish and treat the cells in four groups:

[0062] (1) Control group, without adding any materials;

[0063] (2) 5 μg / mL group, add 16 μL of the BMPR prepared in Example 1 to 984 μL of DMEM medium, and the concentration of Mn is 5 μg / mL;

[0064] (3) 10 μg / mL group, add 32 μL of the BMPR prepared in Example 1 to 968 μL of DMEM medium, and the concentration of Mn is 10 μg / mL;

[0065] After incubating in an incubator at 37°C and 5% CO 2 for 24 h, collect the cells in each group, extract the total cellular proteins using a kit, and perform protein quantification according to the BCA reagent instructions. After electrophoresis, transfer the separated proteins to a PVDF membrane. After transferring the membrane, block it with skim milk for 1 h. Using GAPDH as the internal reference, add the primary antibodies against CollgenⅠ, HIF-1α, Smad4, TGF-β1, and α-SMA diluted according to the instructions. Incubate at 4°C for 14 h. After washing with PBST, add the secondary antibody and gently shake on a shaker for 1 h, wash with PBST, develop, and take pictures to analyze the results of the WB experiment.

[0066] The results are as Figure 4 shown, indicating that the targeted BMPR nanomaterial has a stronger ability to inhibit the expression of each protein in the TGF-β1 / Smad pathway than the non-targeted BMP nanomaterial and can inhibit the activation of hepatic stellate cells.

[0067] Test Example 5

[0068] Examine the cytotoxicity of the BMPR prepared in Example 1. The specific steps are as follows:

[0069] Prepare a 96-well plate, add 1×10 4 of LX-2 cells to each well, culture in a 37°C constant temperature incubator containing 5% CO 2 for 24 h, and then add the BMPR nanomaterial prepared in Example 1 to make Mn 2+The concentrations were 0, 1.25, 2.5, 5, and 10 μg / mL respectively. After mixing evenly, continue to incubate for 24 h, then aspirate the culture medium. Add the culture medium containing MTT to each well and culture for 3.5 h. Aspirate the MTT culture medium, add 150 μL of DMSO to each well, shake on a shaker for 10 min to completely dissolve the purple solid, and use an enzyme-linked immunosorbent assay (ELISA) reader to read the absorbance value (OD value) at 490 nm. Based on the measured OD value, judge the number of viable cells.

[0070] The cell survival rate graph is as Figure 5 shown. As the concentration of the BMPR nanomaterial increases, the survival rate of LX-2 cells decreases significantly, indicating that the BMPR preparation has obvious cytotoxicity to hepatic stellate cells.

[0071] Experimental Example 6

[0072] Investigate the magnetic resonance imaging ability of the BMPR prepared in Example 1. The specific steps are as follows:

[0073] Take 6-week-old healthy and liver fibrosis C57 male mice, and inject the BMPR prepared in Example 1 into the mice via the tail vein at a drug concentration of 0.02 mM / kg of Mn 2+ Conduct scans using a 9.4T small animal nuclear magnetic resonance instrument before injection, 0.5 h after injection, 1 h after injection, 1.5 h after injection, and 2 h after injection, and then perform quantitative analysis on the images.

[0074] The results are as Figure 6 shown. After injecting the nano-preparation, the signal-to-noise ratio of the image is significantly higher than that before injection, and the imaging effect of liver fibrosis mice is significantly better than that of normal mice, indicating that the BMPR nanomaterial has strong magnetic resonance imaging ability and can achieve the effect of monitoring liver fibrosis.

[0075] Therefore, the BMPR nanoprobe prepared by the present invention has good hepatic stellate cell targeting ability, and the targeting effect is enhanced with the increase of the BMPR nanoprobe concentration and the extension of the incubation time; and the higher the severity of liver fibrosis, the more obvious the targeting effect; it has good relaxation performance; it has obvious cytotoxicity to hepatic stellate cells and strong magnetic resonance imaging ability, and can be used for the development of monitoring liver fibrosis and liver fibrosis treatment agents; it can effectively inhibit the expression of each protein in the HIF-1α / TGF-β1 / Smad pathway, thereby inhibiting the activation of hepatic stellate cells, effectively improving the hypoxic microenvironment of liver fibrosis, and has excellent therapeutic effects on liver fibrosis; the provided preparation method is simple and has strong operability; the synthesized product has high stability and good repeatability.

[0076] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit them. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions of the present invention or make equivalent replacements, and these modifications or equivalent replacements cannot make the modified technical solutions deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A BMPR nanoprobe targeting liver fibrosis, characterized in that: It is prepared from bovine serum albumin, pentacarbonyl manganese bromide, dopamine, and arginine-glycine-aspartic acid tripeptide.

2. A BMPR nanoprobe targeting liver fibrosis according to claim 1, characterized in that: The dopamine is maleimide polyethylene glycol dopamine and dopamine hydrochloride.

3. A method for preparing a BMPR nanoprobe targeting liver fibrosis as claimed in claim 2, characterized in that: Here are the steps: S1. Dissolve bovine serum albumin in deionized water, add pentacarbonyl manganese bromide and stir overnight, centrifuge and discard the precipitate to obtain BSA@Mn(CO)5Br solution; S2, adding arginine-glycine-aspartic acid tripeptide and maleimide polyethylene glycol dopamine into deionized water, stirring in an ice bath away from light to obtain a mixed solution; S3. Dissolve the mixed solution obtained in step S2 and dopamine hydrochloride in Tris HCl, mix well and add to the BSA@Mn(CO)5Br solution prepared in step S1, stir for 12 hours in a dark environment, centrifuge to discard the precipitate, dialyze and ultrafilter.

4. The method for preparing a BMPR nanoprobe targeting liver fibrosis according to claim 3, characterized in that: In the step S1, the mass volume ratio of bovine serum albumin: manganese bromide pentacarbonyl: deionized water is 40 mg: 1 mg: 4 mL; and the centrifugation condition is 3000 rpm and centrifugation for 5 min.

5. The method for preparing a BMPR nanoprobe targeting liver fibrosis according to claim 3, characterized in that: In the step S2, the mass volume ratio of arginine-glycine-aspartic acid tripeptide: maleimide polyethylene glycol dopamine: deionized water is 1 mg: 4 mg: 0.5 mL; and the stirring conditions are 400 rpm and 30 min.

6. The method for preparing a BMPR nanoprobe targeting liver fibrosis according to claim 3, characterized in that: The mass volume ratio of maleimide polyethylene glycol dopamine: dopamine hydrochloride: Tris HCl is 3 mg: 12 mg: 2 mL; the pH of Tris HCl is 8.

7. The method for preparing a BMPR nanoprobe targeting liver fibrosis according to claim 3, characterized in that: The centrifugation condition in step S3 is 3000 rpm and centrifugation for 5 min; deionized water is used as the dialysate for dialysis, and the dialysis bag specifications are 44 mm and 14 kDa.

8. Use of the BMPR nanoprobe targeting liver fibrosis as claimed in claim 1 in the preparation of an integrated diagnostic and therapeutic preparation.

9. Use of the BMPR nanoprobe targeting liver fibrosis as claimed in claim 1 in the preparation of a liver fibrosis diagnostic preparation.

10. Use of the BMPR nanoprobe targeting liver fibrosis as claimed in claim 1 in preparing a therapeutic preparation for liver fibrosis.

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