CN111298015A

CN111298015A - Application of Shenmai injection in promoting normalization of blood vessel

Info

Publication number: CN111298015A
Application number: CN202010141918.3A
Authority: CN
Inventors: 周芳; 王广基; 姜超; 王木兰
Original assignee: China Pharmaceutical University
Current assignee: CHIATAI QINGCHUNBAO PHARMACEUTICAL Co.,Ltd.; China Pharmaceutical University
Priority date: 2020-03-04
Filing date: 2020-03-04
Publication date: 2020-06-19

Abstract

The invention belongs to a newly discovered application, relates to the field of new application of medicaments, and particularly relates to a new application of a Shenmai injection, which can improve the tumor vascular structure, reduce the number of vascular branches, increase the vascular maturity, normalize tumor blood vessels, increase the delivery efficiency of 5-FU into tumors and enhance the anti-tumor effect of 5-FU.

Description

Application of Shenmai injection in promoting normalization of blood vessel

Technical Field

The invention relates to the field of medicines, in particular to application of a Shenmai injection in promoting normalization of blood vessels.

Background

The Shenmai injection is a Chinese medicine injection, is a pulse-activating decoction originated from the 'Bingmai' prescription, is prepared by purifying ginseng and ophiopogon root, and mainly comprises ginsenoside, ophiopogonin, ophiopogonone, trace ginseng polysaccharide and ophiopogonpolysaccharide. Has the main effects of tonifying qi, relieving depletion, nourishing yin, promoting fluid production and increasing blood pressure. The traditional Chinese medicine composition is widely used for treating patients with coronary heart disease, myocardial infarction, cardiomyopathy, pulmonary heart disease, heart failure, angina, granulocytopenia, breast cancer, diabetes and the like in clinic. Also can be used for the rescue treatment of critical patients.

Tumors (tumors) are new organisms (neograwth) formed by local tissue cell proliferation of the body under the action of various tumorigenic factors, because such new organisms are mostly in the form of space-occupying block-shaped processes, also called neoplasms (neoplasms). According to the cellular characteristics of the new organism and the degree of harm to the organism, tumors are divided into two major categories, namely benign tumors and malignant tumors. Malignant tumors can be classified into carcinomas and sarcomas, and carcinomas refer to malignant tumors derived from epithelial tissues. Sarcoma refers to malignant tumor of mesenchymal tissue including fibrous connective tissue, fat, muscle, vessel, bone and cartilage tissue, such as carcinoma formed by large intestine mucosa epithelium, which is called large intestine mucosa epithelial cancer, and is called large intestine cancer for short. The skin epithelium is called skin epithelial cancer, which is called skin cancer for short.

Folkman first proposed the "tumor growth is dependent on angiogenesis" hypothesis in 1971, and thus, studies on the regulatory mechanism of tumor angiogenesis and anti-angiogenesis were widely focused. Tumor cells secrete a variety of pro-angiogenic factors such as VEGF, etc., forming a new vascular system, thereby transporting nutrients and oxygen throughout the tumor to meet the needs of growth, proliferation and metastasis. However, the resulting vascular system is very different from normal blood vessels, either morphologically or functionally. Structural abnormalities cause uneven blood flow distribution and high intercellular substance pressure inside the tumor, thereby reducing the delivery efficiency of the antitumor drug into the tumor and affecting the uniform distribution of the drug in the tumor. In 2001, the Jain professor proposed a new idea: the anti-angiogenesis medicine is reasonably applied to enable the original abnormal twisted tumor blood vessels to tend to be normal, thereby more effectively delivering oxygen and the medicine to tumor cells and improving the treatment effect of chemotherapy and radiotherapy. The existing research also shows that part of angiogenesis inhibitors can cause the normalization of tumor blood vessels and tumor micro-rings and improve the treatment effect of radiotherapy and chemotherapy. Normalization of blood vessels is not an immediate effect after administration, nor is it a permanent phenomenon that does not disappear after occurrence, it is a transient, reversible process that occurs at some stage of the anti-angiogenic therapy, which is referred to by researchers as the "time window for normalization of blood vessels" within which normalization of tumor blood vessels occurs structurally and functionally.

The relationship between Shenmai injection and blood vessel, the relationship between Shenmai injection and tumor, and the relationship between Shenmai injection, blood vessel and tumor are not reported at present.

Disclosure of Invention

Based on the above problems, the invention provides an application of Shenmai injection in promoting normalization of blood vessels.

Further, the application of the Shenmai injection in promoting the normalization of blood vessels in colon cancer.

Furthermore, the Shenmai injection is applied to improving the tumor vascular structure, reducing the number of vascular branches, increasing the maturity of blood vessels, normalizing tumor blood vessels, increasing the delivery efficiency of 5-FU into tumors and enhancing the anti-tumor effect of 5-FU.

Furthermore, the Shenmai injection and the 5-FU are combined to inhibit the growth of tumors, increase the tumor inhibition rate of the 5-FU and reduce the weight of tumor tissues; aggravate pathological damage induced by 5-FU, reduce Ki67 expression in tumor tissue, inhibit tumor cell proliferation, and increase 5-FU intratumoral drug concentration.

Furthermore, the Shenmai injection can be used for down-regulating tumor tissue angiopoietin and up-regulating the gene expression level of the angiokine.

Furthermore, the key factors for opening the normalization window period of the Shenmai injection are VEGF, FGF and Angiostatin; the Shenmai injection can up-regulate the enzymatic activities of tPA and uPA in HUVEC cells by inhibiting the expression level of PAI-1 in HUVEC cells of tumor tissues, promote the hydrolysis process of plasminogen and increase the content of Angiostatin in the tumor tissues; inhibiting histone acetylation level of tumor tissue, and down-regulating expression of downstream genes VEGF and FGF to reduce expression level of VEGF and FGF.

Furthermore, the Shenmai injection is combined with 5-FU to increase the anti-tumor effect of the 5-FU and inhibit the growth of tumor.

The invention also provides the application of the Shenmai injection in combination with the promotion of normalization of blood vessels.

Furthermore, the Shenmai injection is combined with 5-FU to inhibit the growth of tumor tissues, inhibit the proliferation of tumor cells and increase the in vivo anti-tumor effect of the 5-FU.

Furthermore, the Shenmai injection combined with 5-FU is applied to reducing the number of blood vessel branches and improving the tumor vascular structure.

Furthermore, the Shenmai injection is combined with 5-FU to increase the positive coverage rate of α -SMA around blood vessels and increase the maturity of the blood vessels.

Further, the method comprises the following steps: the Shenmai injection is used in combination with 5-FU in regulating the balance of blood vessel promoting/inhibiting factors in tumor and promoting normalization of tumor blood vessel.

Furthermore, the Shenmai injection is combined with 5-FU to down-regulate the protein secretion level of VEGF and FGF, up-regulate the protein amount of Angiostatin and increase the delivery efficiency of 5-FU into tumor.

Has the advantages that: the Shenmai injection can improve tumor vascular structure, reduce blood vessel branch number, increase blood vessel maturity, normalize tumor blood vessel, increase 5-FU delivery efficiency into tumor, and enhance anti-tumor effect of 5-FU.

Drawings

FIG. 1 is a graph of the effect of SMI on tumor vasculature at various times after treatment;

FIG. 2 is a graph showing the effect of Shenmai injection on blood flow of tumor tissues at different time nodes;

FIG. 3 is a graph showing the change in tumor vascular maturation after day 10 and day 17 of treatment with Shenmai;

FIG. 4 is a graph of the effect of SMI on the concentration of 5-FU at various times after treatment of a tumor patient;

FIG. 5 is a graph of the synergy of SMI and 5-FU on a LoVo colon cancer xenograft model;

FIG. 6 is a graph of the effect of SMI on tumor vasculature;

FIG. 7 is a graph showing the effect of SMI on tumor angiogenesis factor expression;

FIG. 8 is a graph showing the effect of SMI on the expression of VEGF (A), FGF (B) and angiostatin (C) proteins in tumor tissue.

Detailed Description

Example (b):

the invention provides an application of Shenmai injection in promoting normalization of blood vessels. Further provides the application of the Shenmai injection in promoting the normalization of blood vessels in colon cancer. Further provides the application of the Shenmai injection in improving the tumor vascular structure, reducing the number of vascular branches, increasing the maturity of blood vessels, normalizing tumor blood vessels, increasing the delivery efficiency of 5-FU into tumors and enhancing the anti-tumor effect of 5-FU.

Further, the combination of the Shenmai injection and the 5-FU is proposed to inhibit the growth of the tumor, increase the tumor inhibition rate of the 5-FU and reduce the weight of the tumor tissue; aggravate pathological damage induced by 5-FU, reduce Ki67 expression in tumor tissue, inhibit tumor cell proliferation, and increase 5-FU intratumoral drug concentration.

Further provides the application of the Shenmai injection in down-regulating the tumor tissue angiopoietin and up-regulating the gene expression level of the angiopoietin.

Further, the key factors for opening the normalization window period of the Shenmai injection are VEGF, FGF and Angiostatin; the Shenmai injection can reduce the expression level of VEGF and FGF by inhibiting tumor and down-regulating the expression of downstream genes VEGF and FGF.

Further provides the application of combining the Shenmai injection and the 5-FU to increase the anti-tumor effect of the 5-FU and inhibit the growth of tumors.

Further provides the application of the Shenmai injection combined with 5-FU in inhibiting the growth of tumor tissues, inhibiting the proliferation of tumor cells and increasing the in vivo anti-tumor effect of the 5-FU.

Further provides the application of the Shenmai injection combined with 5-FU in reducing the number of blood vessel branches and improving the tumor vascular structure.

Further provides the application of the Shenmai injection in combination with 5-FU in increasing the positive coverage rate of α -SMA around blood vessels and increasing the maturity of the blood vessels.

Further provides the application of the Shenmai injection combined with 5-FU in regulating the balance of the promotion/inhibition of blood vessel factors in tumors and promoting the normalization of tumor blood vessels.

Further provides the application of the Shenmai injection combined with 5-FU in down-regulating the protein secretion level of VEGF and FGF, up-regulating the protein amount of Angiostatin and increasing the delivery efficiency of 5-FU into tumor.

In order to prove the new application of the Shenmai injection in promoting the normalization of blood vessels, the experimental data are provided as follows:

1. experiment one

Description of the experiment: randomly grouping SPF male LoVo human colon cancer Balb/c mice, injecting different drugs, and observing.

SPF grade male LoVo human colon carcinoma Balb/c mice (9 weeks, 18-22g) purchased from Shanghai Slek laboratory animals, Inc. (Shanghai, China). The experimental animals are adaptively raised in SPF-level special animal rooms of Chinese pharmaceutical university for 3 days. During the adaptive feeding period, water is freely fed and drunk, and a 12-hour day-night circulation is ensured. All experimental animals were in accordance with the requirements of the declaration of helsinki and were approved by the animal ethics committee of the university of chinese pharmacy.

The mice were randomly divided into the following 5 groups:

control group: the control group was injected with normal saline (0.1mL/10g) intraperitoneally every day, and was injected with normal saline (0.05mL/10g) intraperitoneally every three days;

SMI group: the SMI group is singly used for carrying out intraperitoneal injection on 10mL/kg (0.1mL/10g) of Shenmai injection every day, and carrying out intraperitoneal injection on physiological saline (0.05mL/10g) every three days;

5-FU group: the 5-FU group is singly injected with normal saline (0.1mL/10g) intraperitoneally every day, and 5-FU15mg/kg (3mg/mL, 0.05mL/10g) is injected intraperitoneally every three days;

group Bv: the bevacizumab-only group was intraperitoneally injected with physiological saline (0.1mL/10g) every day, and bevacizumab 5mg/kg (0.5mg/mL, 0.05mL/10g) every three days;

5-FU + SMI group: the combination group of SMI and 5-FU is used for intraperitoneally injecting Shenmai injection 10mL/kg (0.1mL/10g) every day, and intraperitoneally injecting 5-FU15mg/kg (3mg/mL, 0.05mL/10g) every three days;

1.1 SMI can inhibit the number of tumor blood vessels and promote the normalization of tumor blood vessel structure

The vessel morphology was observed by in vivo two-photon microscopy imaging.

After 15 days of drug treatment, 5 mice were selected for each group and FITC-Dextran (70kDa,10mg/mL) was injected into the tail vein. Circulating for 10min, carrying out anesthesia (10mg/mL, 0.05mL/10g) on pentobarbital sodium injected into abdominal cavity of the tumor-bearing mouse, shearing skin of the tumor part of the tumor-bearing mouse to expose the tumor tissue, fixing, and observing the blood vessel shape on a two-photon microscope stage. The wavelength of the exciting light is chosen to be 860nm, the three-dimensional distribution of the tumor microvasculature is observed by utilizing Z-axis scanning, the scanning step diameter is 2 mu m, and the scanning depth is 300 mu m. The effect of figure 1SMI on tumor vasculature at different times after treatment was plotted.

From FIG. 1, it can be seen that the number of tumor vessels in the control group increases with the number of growing days, and the number of branches gradually increases, and the walking pattern becomes more disordered. Bevacizumab (Bv) can obviously inhibit the number of tumor blood vessels, and has a relatively obvious inhibiting effect on the 5 th day of administration, the arrangement of the blood vessels is more regular than that of a control group, the blood vessel branches are further reduced by the 10 th day, only individual relatively thick main blood vessels are left, and the blood vessels are further inhibited by 15 days after administration, and the conventional relatively regular blood vessel network begins to be loosely decomposed; on day 17 most of the vessels were inhibited, the vessels disintegrated in large numbers and the normal network disappeared. Thus, both time points on day 5 and day 10 should be within the window of Bv promoting tumor vascular normalization, and 15 days of administration of the drug have begun to reduce vascular normalization, but the window of Bv at the doses and animal models studied needs to be explored further. The number of tumor blood vessels in the SMI group alone does not change significantly with the administration time, but the blood vessel density at each time point is lower than that of the control group on the whole, and the structure is more regular and has fewer branches than that of the control group. The 5-FU group tumor vascular network is always in a disordered arrangement state, and the blood vessel density increases along with time and tends to increase; after combination of SMI, the number of blood vessels was significantly reduced at the time points of 10 days and 15 days, and the structure was significantly improved, the branching was significantly reduced, and the normalization effect was weakened by the late stage of administration. The window period for normalization of SMI pro-vascularisation is presumed to be around 10-15 days.

1.2 SMI can promote the tumor vascular structure to become regular and increase the blood perfusion

Fig. 2 is a graph of the effect of Shenmai injection on tumor tissue blood flow at different time nodes, where (A) tumor blood flow was measured with a laser

speckle contrast imager

10 and 17 days after treatment, respectively, and the tumor area was marked with a dashed line (B) blood flow was calculated by the imager's analysis software, ＊ p <0.05, and n is 6.

As can be seen, the tumor vasculature became regular during the window of tumor vessel normalization and the amount of blood perfusion increased. Based on the window period presumed by the normalization of the SMI-promoted blood vessels, the blood flow of the tumor microvessels of the tumor-bearing mice was measured by laser speckle imaging on the 10 th day (in the window period) and on the 17 th day (out of the window period), respectively. As can be seen from FIG. 2, at the time point of day 10, there was a tendency for the tumor area blood flow to increase with SMI alone as compared with the control group, and the blood flow was significantly increased with SMI in combination with 5-FU and 5-FU alone; and it can be seen from the figure that the groups given SMIs, either alone or in combination, have a more uniform blood flow distribution. On day 17 after administration, the effect of SMI on regulating blood flow disappeared and there was no significant effect on blood flow distribution.

1.3, SMI can promote the maturity of blood vessels

Fig. 3 is a graph of changes in tumor vascular maturation after day 10 and day 17 of shenmai treatment tumor vessels were immunostained with CD31 (FITC-conjugated, green) and pericyte α -SMA (Alexa Flour 680-conjugated secondary antibody, red) and the ratio of α -SMA positive area to CD31 positive area x 400, n 6 was calculated.

As shown in FIG. 3, the tumor vascular maturation was further examined at day 10 and day 17, respectively, on day 10 after the administration, the ratios of α -SMA positivity to CD31 positivity were 25.59. + -. 6.46%, 26.42. + -. 6.96%, 39.07. + -. 5.98%, 39.33. + -. 4.84%, 41.06. + -. 4.87% and 38.93. + -. 6.86% in the control group, 5-FU group, SMI group, Bv group, 5-FU + SMI group and 5-FU + Bv group, respectively, and the positive coverage around the blood vessels of α -SMA and the vascular maturation were significantly increased and the vascular maturation was increased, at day 17 after the administration, the ratios of α -CD 31 in the control group, 5-FU group, SMI group, Bv group, 5-FU + SMI group and 5-FU + Bv group were 20.59%, 21.42%, 24.90% and 22.3.3%, respectively, and the effects on the tumor were decreased by day 10, 22.59. + -. 6.59%, 26, 3, 24.42, 24, and 22.32, respectively.

1.4 combination of SMI and 5-FU drug to increase the intratumoral drug concentration of 5-FU

Figure 4 is a graph of the effect of SMI on 5-FU concentration at different times after treatment of tumor patients, with 5-FU concentrations being measured at 5,10,15, and 17 days post-treatment, p <0.01, p <0.001, and n-6, respectively.

The drug concentration in the tumors was measured for 5,10,15, and 17 days, respectively. Results as shown in 4, SMI had no significant effect on the 5-FU intratumoral drug concentration on

days

5 and 17, suggesting that these two time points may not be within the time window for pro-vascular normalization of SMI; on day 10 and day 15, SMI can obviously increase the intratumoral drug concentration of 5-FU, the lifting amplitude is most obvious on day 10, the 5-FU drug concentration of the SMI combination group on day 10 and day 15 is respectively 1.90 times and 1.45 times that of the 5-FU single group, and the two time points are within the time window of promoting the blood vessel normalization of SMI. The results of the intratumoral drug concentration measurements matched the previously presumed results of the normalization window period.

2. Experiment two

Mice were randomized into 6 groups and given for 15 days with 12 hours of fasting prior to the last dose.

SM group: the SMI group is singly used for carrying out intraperitoneal injection on 10mL/kg (0.1mL/10g) of Shenmai injection every day, and carrying out intraperitoneal injection on physiological saline (0.05mL/10g) every three days;

5-FU + Bv set: combination of bevacizumab with 5-FU groups daily i.p. saline (0.1mL/10g), every three days i.p. bevacizumab 5mg/kg (0.5mg/mL, 0.05mL/10g) and 5-FU15mg/kg (3mg/mL, 0.05mL/10 g).

2.1, the combination of SMI and 5-FU can obviously increase the anti-tumor effect of 5-FU and inhibit the growth of tumor.

FIG. 5 is a graph of the synergy of SMI with 5-FU on a LoVo colon cancer xenograft model, mice injected intraperitoneally with 5-FU (15mg/kg, every 3 days) with or without SMI (10mL/g, daily) for 15 days. (A) Photographs of tumors from each group were excised on day 15. (B) Tumor growth curves for each group during treatment. P <0.05, P < 0.01. n is 5.

As shown in FIG. 5, compared with the normal saline group, the tumor inhibiting effect is not obvious when SMI is used alone, the tumor inhibiting effect is certain when 5-FU is used alone, and the tumor volume of the SMI and 5-FU combined group is obviously reduced. The long diameter (a) and short diameter (b) of the tumor tissue were measured every two days during the administration using the formula V ═ 1/2 × a × b²Calculating the size of the tumor volume, calculating the tumor volume, and drawing a tumor volume growth curve as shown in the figure. On the last day of administration, the sizes of tumor tissues in the saline group, SMI alone group, 5-FU alone group and SMI and 5-FU combined group were calculated to be 592.48 + -38.32 mm3, 567.91 + -54.96 mm3, 478.26 + -44.01 mm3 and 424.31 + -29.16 mm3, respectively. The volume tumor inhibition rates of different administration groups are calculated by using the data, and the volume tumor inhibition rates of the normal saline group, the single SMI group, the single 5-FU group and the combination group of the SMI and the 5-FU are respectively 0%, 4.15%, 19.28% and 28.38%.

Therefore, on a LoVo colon cancer tumor-bearing mouse model, SMI alone has no obvious tumor inhibition effect, but the combined administration of SMI can obviously increase the anti-tumor effect of 5-FU and inhibit the growth of tumors.

2.2 combination of SMI and 5-FU can improve tumor vascular structure

Fig. 6 is a graph showing the effect of SMI on tumor vasculature, observed by in vivo two-photon microscopy. In order to visually observe the influence of SMI on the micro-vascular structure of tumor tissues of tumor-bearing mice, the change condition of tumor vessels of each administration group is observed by using an in-vivo two-photon microscopic imaging technology. The images represent a 3D reconstruction (600 × 600 × 300mm) acquired by MPLSM. Scale bar 100 μm, n 6.

From fig. 6, it can be observed that the tumor microvessels in the control group have high density, different diameters, irregular shapes, disordered blood vessels, disordered walking, a plurality of obvious fine branches, and thick blood vessels in a swelling and blocking state; similarly, the tumor vessels of the 5-FU single-use group also present abnormal states, different forms, uneven distribution and obvious branching and truncation; the density of the microvasculature of the positive drug bevacizumab group is obviously reduced, the blood vessel branches are reduced, and the disfigurement degree is obviously improved; although the density of the tumor microvessels of the SMI group is lower than that of the bevaco group, the density of the tumor microvessels is obviously lower than that of the control group and that of the 5-FU group, the number of blood vessel branches is reduced, the distribution tends to be uniform, and the shape is regular; compared with the combined group of 5-FU and SMI and the combined group of 5-FU and Bv and the single group of 5-FU, the blood vessel density is obviously reduced, the branches are obviously reduced, the shape trend is regular, and the improvement effect of bevacizumab on the blood vessel is stronger than that of SMI. The experimental results show that SMI can improve the tumor vascular structure.

2.3 Combined use of Ginseng and Mai-Mai 5-FU can promote/inhibit the expression level of angiogenesis factor gene in tumor

Figure 7 is a graph of the effect of SMI on tumor angiogenesis factor expression, measured for pro-and anti-angiogenic factor gene expression, P <0.05, P <0.01, P <0.001, n-6.

In the process of tumorigenesis, angiogenesis promoting factors are over-expressed, so that the content of angiogenesis promoting/inhibiting factors is in an unbalanced state, and further tumor development and metastasis are promoted, therefore, whether SMI can regulate the unbalanced state between angiogenesis promoting/inhibiting factors, thereby inhibiting tumor angiogenesis and promoting tumor vessel normalization is examined, and the expression levels of factors closely related to angiogenesis in tumor tissues are measured by a PCR method, wherein 10 of the angiogenesis promoting factors are angiogenesis promoting factors, as shown in FIG. 7, SMI remarkably down-regulates the gene expression levels of most of the angiogenesis promoting factors including HIF-1 α -2 α 2, FGF and ANG, and simultaneously remarkably up-regulates the gene expression levels of part of the angiogenesis inhibiting factors including TIMP1, TIMP2 and Angiostain, compared with the 5-FU single use group.

2.4, the 5-FU combined with Shenmai can reduce the protein secretion level of VEGF and FGF, and obviously up-regulate the protein amount of Angiostatin.

Among the angiopoietins and angiostatic factors, three factors, VEGF, FGF and Angiostatin, have high expression levels in tumor tissues and are significantly regulated by SMI. The protein levels of these three factors were further measured by ELISA and showed that SMI significantly down-regulated the protein secretion levels of VEGF and FGF and significantly up-regulated the amount of Angiostatin protein (as shown in fig. 8). The results were consistent with those of the gene assay.

Claims

1. Application of SHENMAI injection in promoting blood vessel normalization is provided.

2. Use according to claim 1, characterized in that: application of SHENMAI injection in promoting normalization of blood vessel in colon cancer is provided.

3. Use according to claim 1, characterized in that: the Shenmai injection is used for improving tumor vascular structure, reducing blood vessel branch number, inhibiting tumor blood vessel number, increasing blood vessel maturity, normalizing tumor blood vessel, increasing delivery efficiency of 5-FU into tumor, and enhancing anti-tumor effect of 5-FU.

4. Use according to claim 3, characterized in that: the Shenmai injection and the 5-FU are combined to inhibit the growth of tumors, increase the tumor inhibition rate of the 5-FU and reduce the weight of tumor tissues; aggravate pathological damage induced by 5-FU, reduce Ki67 expression in tumor tissue, inhibit tumor cell proliferation, and increase 5-FU intratumoral drug concentration.

5. Use according to claim 1, characterized in that: the Shenmai injection can be used for down-regulating tumor tissue angiopoietin and up-regulating the gene expression level of angiokine.

6. Use according to claim 1, characterized in that: the key factors for opening the normalization window period of the Shenmai injection are VEGF, FGF and Angiostatin; the Shenmai injection can reduce the expression level of VEGF and FGF by inhibiting tumor and down-regulating the expression of downstream genes VEGF and FGF.

7. Use according to claim 1, characterized in that: the Shenmai injection is combined with 5-FU to increase the anti-tumor effect of the 5-FU and inhibit the growth of tumor.

8. Use according to claim 1, characterized in that: the Shenmai injection is combined with 5-FU to inhibit the growth of tumor tissues, inhibit the proliferation of tumor cells and increase the in vivo anti-tumor effect of the 5-FU.

9. Use according to claim 1, characterized in that: the Shenmai injection combined with 5-FU is used for reducing the number of blood vessel branches and improving the tumor vascular structure.

10. The use of claim 1, wherein the Shenmai injection is used in combination with 5-FU to increase α -SMA positive coverage around blood vessels and increase vascular maturation.

11. Use according to claim 1, characterized in that: the Shenmai injection is used in combination with 5-FU in regulating the balance of blood vessel promoting/inhibiting factors in tumor and promoting normalization of tumor blood vessel.

12. Use according to claim 1, characterized in that: the Shenmai injection is combined with 5-FU to down-regulate the protein secretion level of VEGF and FGF, up-regulate the protein amount of Angiostatin and increase the delivery efficiency of 5-FU into tumor.