The subject is a key project in Heilongjiang Province (No. 20030101)
Owner: Surgery Department of the Second Affiliated Hospital of Harbin Medical University 150086
Corresponding author: Fu Zhaochen 0451-86605843
Changes and significance of VEGF after local injection of Xiaozhiling in treatment of hemangioma
Fu Zhaochen Wang Taihe Wang Xiaofeng
【Abstract】 Objective: To find an objective index to reflect the clinical effect of hemangioma patients after local injection of Xiaozhiling, and to explore the cause of vascular endothelial growth
The relationship between VEGF and the pathogenesis of hemangioma. Methods: The immunoenzyme-labeling technique was used to detect 10 normal people and 10 hemangioma patients for treatment
The concentration of VEGF in peripheral blood serum of 8 patients before, 1 week after local injection of Xiaozhiling and 1 month later. Results: In the serum of 10 patients with hemangioma
The VEGF concentration was significantly higher than that of the normal group (P<0.01); the serum VEGF concentration was compared with the hemangioma patient group after 1 week of local injection of Xiaozhiling
Significant difference (P>0.01); 8 patients with hemangioma received local injection of Xiaozhiling for 1 month, and the serum VEGF concentration in peripheral blood was not significantly different from the normal group
Don't (P>0.01). Conclusion: VEGF can be used as an objective basis to reflect the clinical therapeutic effect and prognosis of local injection of Xiaozhiling method; hemangioma
The clinical effect of local injection of Xiaozhiling in patients is definite.
[Keywords] hemangioma; vascular endothelial growth factor; Xiaozhiling; enzyme-linked immunosorbent assay
Serum VEGF level of the patientsw ith hemangioma before and after X iaozhiling injection
Fu Zhaochen,WangTaihe,WangXiaofeng.Department ofOral andMaxillofacialSurgery,The Sec-
ondAffiliatedHospital ofHarbinMedicalUniversity, Harbin150086,China
Abstract: Objective: To observe the effects of Xiaozhiling on the serum vascular endothelialgrowth factor(VEGF)
level of the patients with hemangioma.M ethods:10 healthy subjects (control group) and 10 hemangioma patients
(treatmentgroup) were included. The serum VEGF level were evaluated by ELISA in both groups. The treatment
group was injected with Xiaozhiling once.VEGF level of the treatment group were measured before, one week after
and onemonth(only 8 patients) afterXiaozhiling injection.Results:The serum VEGF concentrations (pg/ml) of the
healty subjects, hemangioma patients without treatment, the patients 1 week after injection of Xiaozhiling and those 1
month after injection were 97. 3±15. 1, 799. 3±84. 1, 828. 1±68. 8 and 105. 6±13. 5 respectively. 1 week after
injection tumor volume decreased bymore than 25% in all the 10 patients.Conclusions:In situinjection of Xiaozhi
lingmay be amethod for the treatment ofhemangioma.
【Key words】 Hemangioma;Vascular endothelialgrowth factor;Xiaozhiling;Euzyme-linked immu-
Chinese Library Classification Number: R739·8 Document Identification Code: A Article Number: 1001-3733(2005)-02-0245-03
The specific pathogenic factors of hemangioma are not very clear. At present, most scholars believe that VEGF is the main factor in the formation of hemangioma. VEGF most
Folkman has long proposed that ischemia and hypoxia can induce the body to produce VEGF, and VEGF is currently considered to be the most effective and in-depth study of blood.
Tube growth factor. Its main biological functions are: (1) Selectively enhance the proliferation of vascular endothelial cells and promote blood vessel formation; (2) Increase blood vessels
In particular, the permeability of tiny blood vessels allows plasma macromolecules to extravasate and deposit in the matrix outside the blood vessels, which contributes to the growth of tumor cells and the construction of neovascular networks.
Provide nutrition immediately. Xiaozhiling has been mostly used in the anorectal department to treat hemorrhoids. Its main ingredients are gallnut, alum, glycerin, chlorobutanol and stabilizers. The mechanism of action is: hemangioma quickly damages vascular endothelial cells after local injection of Xiaozhiling, causing erythrocytes to agglutinate, protein coagulation and thrombosis, resulting in luminal occlusion and tissue fibrosis. In this experiment, the concentration of VEGF in peripheral blood serum of normal people and patients with hemangioma were measured before treatment, 1 week after local injection of Xiaozhiling and 1 month after the immunoenzyme labeling technology, to explore the relationship between VEGF and the incidence of hemangioma, and determine the local The curative effect of Xiaozhiling injection in the treatment of hemangioma, and provides a basis for the clinical evaluation of various treatment methods for hemangioma in the future.
1 Materials and methods
1. 1 specimen collection
·245·J PractStomatol (J PractStomatol) 2005Mar, 21(2) 10 healthy normal people were randomly selected as the normal group; 10 cases of blood
Tumor patients (all are cavernous hemangioma, including 4 cases of lower lip hemangioma, 3 cases of upper lip hemangioma, 3 cases of buccal hemangioma; 2 of them were operated
There was no follow-up one month later. ) Are patients from the Department of Oral Surgery, the Second Affiliated Hospital of Harbin Medical University. The injection method is 5 ml after routine disinfection
The disposable syringe sucks the Zhiling injection, and inserts the needle on the surface of the tumor body, and the dose is injected until the tumor body is slightly pale. The above 10 patients with hemangioma were all treated by local injection of Xiaozhiling once, followed by a follow-up visit and blood samples before the operation, one week after the operation, and one month later. The specimen collection method is fasting, from
4 ml blood (without anticoagulant) is drawn from the peripheral vein and placed in a centrifuge tube, and stored in a refrigerator at -80℃ for later use.
1. 2 Experimental method
Using the ELISA method, the human VEGF kit is a product of Shenzhen Jingmei Biologics Co., Ltd. The method is the double antibody sandwich method. The operation steps are as follows:
Dilute the washing solution 1:20 times with deionized water for use; standard configuration: use a 1,000 μl sampler to take 1.0 ml standard/specimen dilution
Inject 20 ng standard substance (concentration of 20 000 pg/ml); let stand at room temperature for 15 minutes, and configure the concentration to 1000,
500, 250, 125, 62.5, 31.25, 0 pg/ml standard concentration reagent; processing of the blood sample to be tested: take the blood sample to be tested out of the -80℃ refrigerator, and after it is completely dissolved at room temperature, use a centrifuge to Centrifuge at a speed of 1,000 r/min for 10 minutes and then set aside (number omitted). Take out the specimen-coated slats, and add 100μl standard or serum to be tested into each hole with a 100μl micropipette according to the above number. Incubate for 90 min in 37℃ incubator, use the prepared
Wash the plate 4 times with washing liquid.
Table 1 ``Quantitative results of VEGF in serum of each group (pg/m, l-x±s)
Tab 1 VEGF levels in differentgroups (pg/m,l-x±s)
Group Number of cases VEGF value
Normal group 10 97.3 + 15. 1
Patients with hemangioma before treatment 10 799.3+84.1
The patient received local injection of Xiaozhiling for 1 week 10 828.1+68.8
The patient received local injection of Xiaozhiling for January 8 105.6+13.5
Add 100μl of biotinylated antibody working solution to each well, cover the reaction well with sealing tape (the working solution is prepared 15 min in advance), and incubate at 37°C
The plate was washed 4 times after 60 min incubation in the incubator; 100μl of enzyme conjugate working solution was added to each well, and the plate was washed 4 times in the 37℃ incubator for 30 min. each
Add 100μl of working solution of substrate color reagent to the well, and incubate for 2min at 37℃ in the dark. Add 100μl of stop solution to each well and mix well, at a wavelength of 450 nm
Measure the absorbance value (completed within 5 minutes); automatically print the result, the result is the absorbance value, converted to pg/m, and the statistical method is a two-sided t test.
Efficacy evaluation criteria: ① Significantly effective means that the tumor body disappears completely, the skin and mucous membranes are normal, no dysfunction, and the postural movement test is negative; ② Effective means that the tumor body is reduced>
25%, but not completely disappeared, the deformity improved, the posture movement test can be negative or positive; ③Ineffective is the tumor shrinkage <25%, or the signs before and after treatment are unknown
Significant improvement. The local shrinkage of the hemangioma in 10 patients with hemangioma was more than 25% when they were revisited a week later, without skin and mucosal damage, and the effective rate was 100%.
The concentration of VEGF in the serum of 10 patients with hemangioma was significantly higher than that in the normal group (P<0.01), and 10 patients with hemangioma were injected locally with Xiaozhiling
There was no significant difference in serum VEGF concentration before treatment after 1 week (P>0.01), which was significantly different from the normal group (P<0.01), 8 cases of blood vessels
There was no significant difference in serum VEGF concentration in peripheral blood of patients with tumor after local injection of Xiaozhiling for 1 month (P>0.01).
2 Discuss Discuss
Hemangioma is a kind of benign tumor and vascular malformation of the skin or subcutaneous tissue with the highest incidence in infants and young children . Clinically
There are many treatment methods for hemangioma, including surgical resection, laser therapy, interferon therapy, hormone therapy, photochemotherapy, and local injection of sclerosing agent therapy. Among them, the local injection therapy of Xiaozhiling is widely used in clinical practice. However, there is a lack of an objective index to reflect the therapeutic effect and prognosis of various treatment methods, which brings difficulties to the clinical treatment of hemangioma. The purpose of this experiment is to find an objective indicator to reflect the treatment effect. The exact pathogenesis of hemangioma is unclear, and most scholars believe that VEGF is a direct factor in the formation of hemangioma, and its continuous high expression is the main factor that determines angiogenesis [2~5]. VEGF is a dimer formed by two identical subunits with a relative molecular mass of 17 000-22 000 connected by disulfide bonds. VEGF is divided into 4 subtypes according to the alternative splicing sites of exons in RNA transcription. : VEGF121, VEGF165, VEGF189, VEGF206[6, 7]. VEGF works as follows: the receptor first autophosphorylates, then activates phosphatidylcholine-specific phospholipase C (PLC-r), PLC-r hydrolyzes phosphatidylinositol diphosphate (PIP2), and produces diacylglycerol (DAG). ) And inositol triphosphate (IP3), among which DAG can activate protein kinase C in the cytoplasm and fix it on the membrane, then induce the growth of endothelial cells and increase vascular permeability.
In this experiment, the VEGF concentration of the hemangioma patient group was significantly higher than that of the normal group (P<0.01), indicating that VEGF in the peripheral blood serum of the hemangioma patient
The concentration of vascular tumor in patients with hemangioma was not significantly different from that before treatment after local injection of Xiaozhiling for 1 week, but was significantly different from that of the normal group.
The difference (P<0.01). This is because VEGF is not only secreted by vascular endothelial cells, but hematopoietic stem cell lines such as platelets, megakaryocytes, and mononuclear cells
Cells and lymphocytes also secrete VEGF. One week after local injection of hemangioma, the body compensatory synthesis and secretion due to stimulation of local oxidants
VEGF, so the concentration in peripheral blood increases. After 1 month, the body's inflammatory response has disappeared, and VEGF is metabolized in the body, so the peripheral blood serum
The concentration of VEGF is close to the normal level. We can conclude from this: the prescription of local injection of Xiaozhiling for hemangioma
·246·J PractStomatol 2005Mar, 21(2) The clinical effect is definite; measuring the concentration of EGF in peripheral blood serum by ELISA method can accurately reflect the concentration of vascular endothelial growth factor in peripheral blood [9~10]; peripheral blood The concentration of VEGF in serum can be used as an effective indicator to judge the clinical treatment effect and prognosis, providing a reference indicator for the comparison of the effects of various treatment methods for patients with hemangioma in the future, and for further scientific research and clinical research. Provide evidence.
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(Received: 2004-6-17 Revised back: 2004-11-15)