CN112933212B

CN112933212B - Application of SND1 in preparation of medicine for preventing and treating vascular injury ischemia

Info

Publication number: CN112933212B
Application number: CN202110372805.9A
Authority: CN
Inventors: 黄恺; 陈敏; 胡霁; 梁明露
Original assignee: Tongji Medical College of Huazhong University of Science and Technology
Current assignee: Tongji Medical College of Huazhong University of Science and Technology
Priority date: 2021-04-07
Filing date: 2021-04-07
Publication date: 2022-05-13
Anticipated expiration: 2041-04-07
Also published as: CN112933212A

Abstract

The invention discloses application of SND1 in preparation of a medicine for preventing and treating vascular injury ischemia. According to the invention, the SND1 can promote endothelial progenitor cells to proliferate, migrate and form tubes, so that angiogenesis is promoted, and vascular diseases caused by ischemia, including lower limb ischemia, can be prevented and/or treated.

Description

Application of SND1 in preparation of medicine for preventing and treating vascular injury ischemia

Technical Field

The invention relates to application of SND1, in particular to application of SND1 in preparation of a medicine for preventing and treating vascular injury ischemia.

Background

Staphylococcal nucleic acid Domain Protein (SND 1, Staphylococcus nuclear And gold Domain binding 1), also known as Staphylococcus nuclear Domain-binding Protein 1, EBNA2 Coactivator P100, And Staphylococcus nuclear Domain-binding Protein 11, comprising 910 amino acids And having a molecular weight of 101997 Da. NCBI reference sequence No. NP-055205.2, amino acid sequence as follows: MASSAQSGGSSGGPAVPTVQRGIIKMVLSGCAIIVRGQPRGGPPPERQINLSNIRAGNLARRAAATQPDAKDTPDEPWAFPAREFLRKKLIGKEVCFTIENKTPQGREYGMIYLGKDTNGENIAESLVAEGLATRREGMRANNPEQNRLSECEEQAKAAKKGMWSEGNGSHTIRDLKYTIENPRHFVDSHHQKPVNAIIEHVRDGSVVRALLLPDYYLVTVMLSGIKCPTFRREADGSETPEPFAAEAKFFTESRLLQRDVQIILESCHNQNILGTILHPNGNITELLLKEGFARCVDWSIAVYTRGAEKLRAAERFAKERRLRIWRDYVAPTANLDQKDKQFVAKVMQVLNADAIVVKLNSGDYKTIHLSSIRPPRLEGENTQDKNKKLRPLYDIPYMFEAREFLRKKLIGKKVNVTVDYIRPASPATETVPAFSERTCATVTIGGINIAEALVSKGLATVIRYRQDDDQRSSHYDELLAAEARAIKNGKGLHSKKEVPIHRVADISGDTQKAKQFLPFLQRAGRSEAVVEYVFSGSRLKLYLPKETCLITFLLAGIECPRGARNLPGLVQEGEPFSEEATLFTKELVLQREVEVEVESMDKAGNFIGWLHIDGANLSVLLVEHALSKVHFTAERSSYYKSLLSAEEAAKQKKEKVWAHYEEQPVEEVMPVLEEKERSASYKPVFVTEITDDLHFYVQDVETGTQLEKLMENMRNDIASHPPVEGSYAPRRGEFCIAKFVDGEWYRARVEKVESPAKIHVFYIDYGNREVLPSTRLGTLSPAFSTRVLPAQATEYAFAFIQVPQDDDARTDAVDSVVRDIQNTQCLLNVEHLSAGCPHVTLQFADSKGDVGLGLVKEGLVMVEVRKEKQFQKVITEYLNAQESAKSARLNLWRYGDFRADDADEFGYSR

The staphylococcal nucleic acid domain protein (SND 1) consists of an N-terminal SN (staphylococcus) domain and a C-terminal TSN (Tudor SN5) domain, wherein the SN domain comprises four repeated functional fragments from SN1 to SN 4. In the existing research, the transcription coactivator interacts with the acidic region of Epstein-Barr virus nuclear antigen 2 (EBNA 2), is a transcription coactivator required by B lymphocyte transformation, and interacts with transcription proteins including signal transduction factors and transcription activator STATs. SND1 is thought to be essential for normal cell growth and is a component of the RNA-induced silencing complex (RISC). SND 1-associated diseases include pancreatic acinar cell adenocarcinomas, with associated pathways including the C-MYB transcription factor network and tumorigenic MAPK signaling pathways, and GO annotation associated with this gene including nucleic acid binding and transcriptional coordinator activity.

Endothelial Cells (ECs) line the lumen of all blood vessels and play a key role in maintaining the barrier function of the vascular system. When the vascular endothelial cells are damaged and ischemic, the inflammation and the function of the endothelial cells are changed, thereby causing the occurrence and the development of various diseases, such as atherosclerosis, pneumonia, vascular calcification, vascular sclerosis and the like. At present, there are several drugs for improving vascular ischemia, the most common of which are: nimodipine, flunarizine, papaverine, etc. The existing research shows that SND1 has nucleic acid binding and transcriptional coordination node activity, and the role of the SND1 in vascular ischemia-related diseases such as limb ischemia is not researched.

Disclosure of Invention

The invention aims to solve the problems in the prior art, and provides application of SND1 in preparing a medicine for preventing and treating vascular injury ischemia.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

application of SND1 in preparing medicine for preventing and treating vascular injury ischemia is provided.

Preferably, the application is the application of SND1 as VEGFR2 and/or ERK signal pathway activator in preparing medicines for preventing and treating vascular injury ischemia.

Preferably, the application is the application of SND1 as an angiogenesis activator in the preparation of medicines for preventing and treating vascular injury ischemia.

Preferably, the vascular injury ischemia is lower limb vascular injury ischemia.

Preferably, the application is the application of SND1 in preparing a medicine for preventing and treating lower limb vascular injury ischemia.

The invention also provides a VEGFR2 signal path activator, which comprises SND1 as an active ingredient.

The invention has the beneficial effects that:

SND1 has activation effect on VEGFR2 signal path and/or ERK signal path, so that SND1 can improve blood vessel ischemia, thereby preventing and/or treating diseases caused by blood vessel ischemia;

SND1 can be widely used as a medicine for treating related diseases in which endothelial cells participate, such as lower limb ischemia, vascular injury and the like.

Drawings

FIG. 1 shows the recovery of mice in the control and treated groups;

FIG. 2 shows SND1 protein expression and relative transcription in lower limbs;

FIG. 3 is a statistical chart of HUVEC proliferation assay in control and treatment groups using EdU assay;

FIG. 4 shows the HUVEC proliferation status of the control group and the treatment group detected by tube formation experiment;

FIG. 5 shows the HUVEC proliferation of the control group and the treatment group detected by the scratch test;

FIG. 6 shows the HUVEC proliferation assay in the control and treatment groups using the Transwell method;

FIG. 7 shows the expression of the VEGF receptor 2 (VEGFR 2) signaling pathway and ERK signaling pathway proteins.

Detailed Description

The technical solution of the present invention will be further described in detail with reference to specific embodiments. It is to be understood that the following examples are only illustrative and explanatory of the present invention and should not be construed as limiting the scope of the present invention. All the technologies realized based on the above-mentioned contents of the present invention are covered in the protection scope of the present invention.

The invention mainly relates to application of SND1 in preparation of a medicament for preventing and treating vascular injury ischemia.

Furthermore, the application is the application of the SND1 as VEGFR2 and/or ERK signal pathway activator in the preparation of the medicine for preventing and treating vascular injury ischemia.

Further, the application is the application of SND1 as an angiogenesis activator in the preparation of medicines for preventing and treating vascular injury ischemia.

Further, the vascular injury ischemia is lower limb vascular injury ischemia.

Further, the application is the application of SND1 in preparing a medicine for preventing and treating lower limb vascular injury ischemia.

The invention also provides a VEGFR2 signal path activator, the active ingredient of which comprises SND1

The invention uses cell experiment to research the promoting effect of the medicine on the proliferation, migration and tube formation of endothelial cells, and VEGF used in the test of the invention is purchased from Kinseri, model Z03073. The proliferation, migration and tube formation of endothelial cells were examined by the following methods.

Unless otherwise indicated, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods.

Wherein and/or when the contents of the scheme for determining protection are indicated, either one or both of the technical features may be selected.

Detecting the proliferation condition of the HUVEC cells by using an EdU cell proliferation experiment, detecting the migration condition of the HUVEC cells by using a cell scratch and transwell experiment, and performing the overexpression of the SND1 protein by using adenovirus transfection. The detection method is described in Huang D, Wang Y, Wang L, Zhang F, Deng S, Wang R, Zhang Y, Huang K. Poly (ADP-rib) polymerase 1 is Induced dependent for transformation growth factor-beta Induced Smad3 activation in vacuum chamber tissue cell, PLoS one 2011, (6) (10) e27123. the tube forming condition of HUVEC cells is detected by applying a tube forming experiment. The detection method is described in Maria Teresta Gentile, Olga Pastorino, Maurizio Bifulco, Luca Colluci-D' Amato. HUVEC Tube-formation Assay to Evaluate the Impact of Natural Products on angiogenesis. J Vis Exp. 2019; (148).

The specific process is as follows:

EdU cell proliferation assay: primary umbilical vein endothelial cells (HUVECs) of human origin were seeded in 96-well plates and cells were treated with siRNA knockdown controls, siRNA knockdown SND1, adenovirus controls and adenovirus supra SND1 for 24 h, respectively. Incorporation analysis of EdU was performed according to the manufacturer's instructions (EdU staining kit (C10310, ribo, guangzhou)), and results were taken with Olympus cellSens Entry.

Cell scratch test: inoculating HUVEC intoIn 6-well plates, culture was performed to 80% density. After the cells were treated with siRNA knockdown control, siRNA knockdown SND1, adenovirus control, and adenovirus watch SND1 for 24 h, respectively, the cell monolayer was scratched with a 200. mu.l pipette tip, approximately every 0.5-1 cm line, and after scratching, the cells were washed 3 times with PBS to remove scratched cells, and then placed in a 37 ℃ 5% CO chamber₂The cells were observed in an incubator, starved for 1 h, stimulated with VEGF (25 ng/ml), and the wound closure rate was measured using the Image J program at 0, 6, 12, and 24 h using an Olympus cellSens entry.

Cell migration was measured by the Transwell method: uniformly smearing 200 μ l rat tail collagen ((C7661, Sigma, 0.5 mg/mL) under the chamber, air drying, placing the chamber into a culture plate, adding 200 μ l of pre-warmed serum-free culture medium into the upper chamber, standing at room temperature for 15-30 min to make the matrix glue hydrated and suck the rest culture solution, inoculating HUVEC into a 6-well plate, culturing to 80% density, treating the cells with siRNA knockdown control, siRNA knockdown SND1, adenovirus control and adenovirus over SND1 for 24 h, sowing the cells into the upper air chamber, culturing in M199 containing 10% fetal calf serum, 500 mL M199 and 10% fetal calf serum and VEGF (25 ng/mL) (the control group is equal volume of DSMO) into the lower air chamber, standing for 12 h, fixing the cells with 4% formaldehyde for 20 min, and (3) dyeing the crystal violet with the mass concentration of 0.1% for 20 min. Migrated cells were photographed using the Olympus cellSens channel.

Adenovirus packaging and transfection: 293 cells were seeded in 1 60 mm culture dish, 6. mu.g of plasmid was transfected into cells by PEI at a cell density of 50-70%, after 6-8 h of culture, the medium was removed, 2-3 ml of DMEM complete medium (FBS at a volume density of 10%) was added, and the cells were blown off by pipette 7 to 10 days after transfection. The solution was collected, centrifuged to discard the supernatant, cells were resuspended in 2.0 ml PBS, frozen in liquid nitrogen, lysed in a 37 ℃ water bath, and shaken vigorously. This step was performed 3 times in total to obtain virus-containing supernatant. 293 cells were plated at 50-70% confluence onto 60 mm plates and virus-containing supernatant was added at 30-50% by volume. Obvious cell lysis or cytopathic effect (CPE) phenomenon can be seen 2-3 days after infection, and virus supernatant is collected again. 3 rounds of amplification were performed to obtain sufficient virus titer. Human primary HUVEC cells were cultured to a cell density of 70-80%, the collected virus was infected into HUVEC cells, and after 24-48 h, cell fluorescence was observed under a microscope.

Adenovirus treatment: the tail vein is injected into the mice once every two weeks, 5X 10 times each time⁸pfu, total 200. mu.l, empty virus as blank control; 30-100 μ l of adenovirus is added to each well of the six-well plate.

Endothelial cell tube formation experiment: 50 μ l of thawed ECM gel (356234, BD) was added to 96-well plates and incubated at 37 ℃ for 30 min, after which HUVECs were seeded into 96-well plates, incubated at 37 ℃ for 4h, the piped cells were photographed using an Olympus cellSens channel, and the results were counted using Image J software.

Mouse lower limb ischemia model experiment: injecting pentobarbital (10 μ l/g mouse weight) with mass volume ratio of 0.5% into abdominal cavity, anesthetizing, depilating left lower limb, and fixing on operation plate; cutting skin from the groin of the left lower limb to fully expose femoral artery and femoral vein, and slightly separating the femoral artery and the femoral vein by using microscopic forceps under a microscope; crossing 2 strands of 6-0 filaments from the lower part of femoral artery, ligating blood vessels at the lower edge of groin and the proximal end of bifurcation of femoral artery, and cutting off the middle blood vessel of two ligation knots by using a micro-scissors; suturing the incision, disinfecting with alcohol cotton ball, and placing on 37 deg.C hot pad for keeping the temperature until the mouse revives; blood flow of the two lower limbs is detected by using blood flow Doppler on the 1 st, 3 rd, 7 th and 14 th days before and after the operation respectively, and the blood flow proportion of the ischemic lower limb relative to the non-ischemic lower limb is calculated.

The following embodiments are combined to specifically describe the application of SND1 in preparing the medicine for preventing and treating vascular injury ischemia.

Example 1

All animal experiments were performed according to standard procedures and approved by the animal ethics committee of the college of peer medical college of science and technology, huazhong. The mice were subjected to ischemia surgery on the left and lower limbs. And (5) detecting the ischemia perfusion recovery condition of the lower limb of the mouse by using a Doppler blood flow meter.

In FIG. 1, A is 8-week-old male C57BL/6 mice, to which control (Ad-GFP) and SND1 hyperepiadenovirus (Ad-SND 1) were administered, and both lower limbs were intramuscularly injected once a day, and left lower limb ischemia was performed on the mice. And (3) detecting the ischemia perfusion recovery condition of the lower limb of the mouse by using a Doppler blood flow instrument before the operation, on the first day after the operation, on the third day after the operation, on the seventh day after the operation and on the fourteenth day after the operation respectively. In FIG. 1, B is the relative value of the perfusion recovery of the left lower limb of the mouse in A.

As can be seen from a and B in fig. 1, compared with the control treated group, the blood perfusion recovery of mice in the table SND1 group was significantly increased on the seventh, tenth and fourteenth days after the operation, and the table SND1 promoted the perfusion recovery of ischemic blood vessels in the mice.

C in FIG. 1 is 8-week-old male C57BL/6 background SND1 knock-out control (SND 1)^flox/floxGroup) mice and endothelial-specific knockout SND1 (SND 1 ECKO) mice. The mice were subjected to ischemia surgery on the left and lower limbs. And (3) detecting the ischemia perfusion recovery condition of the lower limb of the mouse by using a Doppler blood flow instrument before the operation, on the first day after the operation, on the third day after the operation, on the seventh day after the operation and on the fourteenth day after the operation respectively. In FIG. 1, D is the relative value of the perfusion recovery of the left lower limb of the mouse C.

As can be seen from C and D in fig. 1, compared with the control treated group, on the seventh, tenth and fourteenth days after the operation, the blood flow perfusion recovery of the SND1 knockout genome mouse is significantly reduced, and the endothelial-specific knockout SND1 inhibits the perfusion recovery of the ischemic blood vessel of the mouse.

Example 2

The left lower limb of an 8-week-old male C57BL/6 mouse is subjected to lower limb ischemia, the mouse is killed on the first day, the third day, the seventh day and the fourteenth day after operation respectively, the tissue of the operation part of the mouse is taken to extract protein, and the protein expression level is detected by adopting protein immunoblotting.

FIG. 2, Panel A is a protein electrophoretogram showing a first behavior of SND1 protein expression and a second behavior of Tubulin protein expression; lanes 1-4 are lower limb histones after the first, third, seventh and fourteenth days of the sham operated group (Non-HLI), respectively, and lanes 5-8 are lower limb histones after the first, third, seventh and fourteenth days of the lower limb ischemia operated group (HLI), respectively.

As can be seen from a in fig. 2, the protein level of SND1 in the lower limb ischemia surgery group was significantly higher than that in the sham surgery group, and the protein expression level of SND1 was significantly increased on the third and seventh days with the increase in post-surgery time, and slightly decreased to the fourteenth day.

In fig. 2B, after different operation time, the lower limb ischemia operation group and the false operation groupsnd1mRNA level Using reference genestubulinNormalized ratio.

As can be seen from B in fig. 2, the mRNA level of SND1 in the lower limb ischemia surgery group was significantly higher on the first, third and seventh days after surgery than in the sham surgery group.

Example 3

HUVEC cells were treated with siRNA knockdown control, siRNA knockdown SND1, adenovirus control, and adenovirus overexpression SND1, respectively, and then tested for HUVEC proliferation using EdU assay.

In FIG. 3, A is siRNA knockdown control group (control group) and siRNA knockdown SND1 treated group (siSND 1) for detecting proliferation of Human Umbilical Vein Endothelial Cells (HUVEC) by EdU assay. Wherein the first column is DAPI stained nuclei, the second column is EdU stained nuclei, and the third column is a fusion map (Merge) of the first two columns; while the first line was the siRNA knockdown control group (control group) and the second line was the siRNA knockdown SND1 treated group (siSND 1).

In FIG. 3, B is a statistical chart of the HUVEC proliferation assay using EdU assay in siRNA knockdown control group (control group) and siRNA knockdown SND1 treated group (siSND 1).

As can be seen from a and B in fig. 3, knock-down of SND1 inhibited HUVEC cell proliferation.

In FIG. 3, C is the HUVEC proliferation assay using EdU assay in adenovirus control group (GFP group) and adenovirus overexpression SND1 treatment group (Ad-SND 1). Wherein the first column is DAPI stained nuclei, the second column is EdU stained nuclei, and the third column is a fusion map (Merge) of the first two columns; while the first behavior was adenovirus control (GFP group) and the second behavior was adenovirus-treated with Table SND1 (Ad-SND 1).

FIG. 3, panel D, is a statistical chart of HUVEC proliferation assay using EdU assay in adenovirus control group (GFP group) and adenovirus overexpression SND1 treatment group (Ad-SND 1).

From C and D in FIG. 3, SND1 was shown to promote HUVEC cell proliferation.

Example 4

FIG. 4 is a graph of HUVEC cell tubulogenesis detected after HUVEC administration of siRNA knockdown control, siRNA knockdown SND1, adenovirus control, and adenovirus overexpression SND1 treatment.

In FIG. 4, A is the siRNA knockdown control group (NC group) and the siRNA knockdown SND1 treated group (si-SND 1) for detecting HUVEC proliferation by tube-forming experiment.

In FIG. 4, B is a statistical chart of the HUVEC proliferation assay using the tube-forming experiment in the siRNA knockdown control group (NC group) and the siRNA knockdown SND1 treated group (si-SND 1).

As can be seen from A and B in FIG. 4, knocking down SND1 inhibited HUVEC cell tubulogenesis.

In FIG. 4, C is the HUVEC proliferation assay in adenovirus control group (GFP group) and adenovirus overexpression SND1 treatment group (Ad-SND 1) using tube-forming experiments.

FIG. 4, D, is a statistical chart of HUVEC proliferation assay using tube-forming experiments in adenovirus control group (GFP group) and adenovirus overexpression SND1 treatment group (Ad-SND 1).

From C and D in FIG. 4, SND1 shows that HUVEC cell tubulation is promoted.

Example 5

HUVEC cells were treated with SiRNA knockdown control, siRNA knockdown SND1, adenovirus control, and adenovirus overexpression SND1, respectively, and then tested for HUVEC cell proliferation and migration ability by a cell scratch test.

In FIG. 5, A is the siRNA knockdown control group (NC group) and the siRNA knockdown SND1 treated group (si-SND 1) for detecting HUVEC proliferation migration by using a scratch test. Wherein the first column is the scratch condition of 0 h, and the second column is the scratch condition after 24 h; while the first row was the siRNA knockdown control group and the second row was the siRNA knockdown SND1 treated group.

In FIG. 5, B is a statistical chart of HUVEC proliferation migration detection by using a scratch test in an siRNA knockdown control group (NC group) and an siRNA knockdown SND1 treatment group (si-SND 1).

As can be seen from A and B in FIG. 5, knocking down SND1 inhibited HUVEC cell migration and proliferation.

FIG. 5, panel C, shows the HUVEC proliferation migration in adenovirus control group (NC group) and adenovirus overexpression SND1 treatment group (Ad-SND 1) by scratch test. Wherein the first column is the scratch condition of 0 h, and the second column is the scratch condition after 24 h; while the first behavior was adenovirus control group and the second behavior was adenovirus cross-table SND1 treated group.

FIG. 5, D is a statistical chart of HUVEC proliferation migration detection by scratch test in adenovirus control group (NC group) and adenovirus overexpression SND1 treatment group (Ad-SND 1).

As can be seen from C and D in fig. 5, knocking down SND1 inhibited HUVEC cell migration and proliferation.

Example 6

HUVEC were treated with siRNA knockdown control, siRNA knockdown SND1, adenovirus control and adenovirus respectively 24 h after surface SND1, and then treated with growth factor VEGF (25 ng/ml) for 12 h, and cell proliferation was detected by the Transwell method.

FIG. 6A shows cell proliferation assay by Transwell method after HUVEC treatment with siRNA knockdown control (NC) and SND1 (si-SND 1) and VEGF (25 ng/ml) for 12 h. The first column is a VEGF treatment group, and the second column is a VEGF treatment group; while the first behavior siRNA knockdown control group, the second behavior siRNA knockdown SND1 treated group;

FIG. 6B is a statistical chart showing cell proliferation assay by the Transwell method after HUVEC were treated with siRNA knockdown control group (NC group) and siRNA knockdown SND1 treated group (si-SND 1), respectively, and then treated with growth factor VEGF (25 ng/ml) for 12 hours;

as can be seen from a and B in fig. 6, knock-down of SND1 significantly inhibited VEGF-induced HUVEC cell proliferation.

FIG. 6C shows the cell proliferation assay by the Transwell method after HUVEC were administered to the adenovirus control group (NC group) and adenovirus treatment group (Ad-SND 1) with Table SND1, respectively, and after VEGF (25 ng/ml) was administered as a growth factor for 12 hours. The first column is a VEGF treatment group, and the second column is a VEGF treatment group; while the first behavior was adenovirus control group and the second behavior was adenovirus cross-table SND1 treated group.

FIG. 6D is a statistical chart showing cell proliferation by the Transwell method after HUVEC was administered to the adenovirus control group (NC group) and the adenovirus treatment group (Ad-SND 1) with Table SND1, respectively, and after 12 hours of VEGF (25 ng/ml) treatment.

As can be seen from C and D in fig. 6, SND1 is shown to further significantly promote VEGF-induced HUVEC cell proliferation.

Example 7

HUVECs are respectively given siRNA knockdown control, siRNA knockdown SND1, adenovirus control and adenovirus, and after 24 h of treatment with SND1, VEGF treatment is carried out, and protein immunoblotting is utilized to detect protein expression conditions of VEGFR signal path and ERK signal path.

In FIG. 7, A shows the expression of VEGF receptor 2 (VEGFR 2) signaling pathway and ERK signaling pathway protein detected by Western blot assay after HUVEC was treated with VEGF (25 ng/ml) for 0, 5, 15 and 30 minutes after siRNA knockdown control (Scramble siRNA) and SND1 (si-SND 1) respectively.

As can be seen from a in fig. 7, VEGF promoted phosphorylation of VEGFR2 and increased phosphorylation of ERK, whereas VEGF promotion was inhibited after knock-down of SND 1.

In FIG. 7, B is the expression of VEGF receptor 2 (VEGFR 2) signaling pathway and ERK signaling pathway protein detected by Western blot assay after HUVEC was administered to adenovirus control group (NC group) and adenovirus overexpression group SND1 treatment group (Ad-SND 1), respectively, and VEGF (25 ng/ml) was used for 0, 5, 15, and 30 minutes.

As can be seen from B in fig. 7, VEGF promoted phosphorylation of VEGFR2 and increased phosphorylation of ERK, whereas VEGF promotion was further activated after passing through table SND 1.

The experimental results show that SND1 can directly activate VEGFR2 signal pathway and ERK signal pathway induced by VEGF, promote HUVEC cell proliferation, migration and tube formation, and improve perfusion recovery of lower limb ischemia.

The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiment. Any modification, equivalent replacement, or improvement made without departing from the spirit and principle of the present invention shall fall within the protection scope of the present invention.

Claims

Application of SND1 in preparing medicines for preventing and treating vascular injury ischemia is disclosed.
2. Use according to claim 1, characterized in that: the application is the application of SND1 as VEGFR2 and/or ERK signal channel activator in the preparation of the medicine for preventing and treating vascular injury ischemia.
3. Use according to claim 1 or 2, characterized in that: the application is the application of SND1 as an angiogenesis activator in the preparation of the medicines for preventing and treating vascular injury ischemia.
4. Use according to claim 1, characterized in that: the blood vessel injury ischemia is lower limb blood vessel injury ischemia.
5. Use according to claim 4, characterized in that: the application is the application of SND1 in preparing the medicine for preventing and treating the vascular injury ischemia of the lower limbs.