Application of PAS-Bu solution in preparation of platelet preservative

Abstract

The invention discloses application of a PAS-Bu solution to preparation of a platelet preservative. The PAS-Bu solution is prepared from sodium butyrate, glucose, sodium chloride, potassium chloride, magnesium chloride, sodium citrate and sodium phosphate. The invention proves that the PAS-Bu solution can effectively relieve activation and apoptosis of platelets, maintain the young state of the platelets, prolong the storage life of the platelets and protect the hemostasis function of the platelets.

Description

Technical Field

[0001] The invention relates to a platelet preservation technology, and in particular to an application of a PAS-Bu solution in preparing a platelet preservative. The PAS-Bu solution is prepared from sodium butyrate, glucose, sodium chloride, potassium chloride, magnesium chloride, sodium citrate and sodium phosphate. Background Art

[0002] Platelets are small pieces of cytoplasm formed by the degranulation of megakaryocytes, and their main function is to participate in coagulation. Although there are many factors that cause the shortage of platelet supply, the short storage period (only 5 days) under conventional storage conditions (22±2℃ continuous shaking) is one of the main factors, which directly leads to an expiration and scrap rate of up to 10% to 20%. It is precisely because of the high expiration and scrap rate, and considering the high price of special equipment and consumables required for platelet collection, it is difficult for blood centers and blood stations to carry out large-scale blood preparation of platelets, resulting in shortages, and the high expiration and scrap rate will further aggravate the shortage. The PAS currently used abroad must be used with plasma in different proportions, allergic reactions cannot be avoided, and the same type of transfusion must be used. Summary of the invention

[0003] In view of the defects or shortcomings of the prior art, the present invention provides an application of a PAS-Bu solution for preparing a platelet preservative; the PAS-Bu solution is prepared from sodium butyrate, glucose, sodium chloride, potassium chloride, magnesium chloride, sodium citrate and sodium phosphate.

[0004] The present invention confirms that the PAS-Bu solution can effectively alleviate platelet activation and apoptosis without adding plasma, while maintaining the resting morphology of platelets, prolonging the shelf life of platelets and protecting the in vivo and in vitro hemostatic function of platelets.

[0005] Optionally, the pH value of the PAS-Bu solution is 6.6-7.4.

[0006] Optionally, the concentration of sodium butyrate in the PAS-Bu solution is greater than or equal to 5 mmol / L.

[0007] Optionally, the concentration of each component in the PAS-Bu solution is: sodium chloride 69.3 mmol / L, potassium chloride 5.0 mmol / L, magnesium chloride 1.5 mmol / L, sodium citrate 10.8 mmol / L, sodium phosphate 26 mmol / L, sodium butyrate 5 mmol / L, glucose 7.5 mmol / L.

[0008] The present invention also provides a corresponding platelet preservative, which is composed of sodium butyrate, glucose, sodium chloride, potassium chloride, magnesium chloride, sodium citrate and sodium phosphate. In some other schemes, the platelet preservative is composed of sodium butyrate, sodium chloride, potassium chloride, magnesium chloride, sodium citrate and sodium phosphate, and glucose is added to the platelet preservative of this scheme before use. The platelet preservative of the present invention is used to preserve platelets, and the preservation time at room temperature is less than or equal to 9 days, such as 5, 7 or 9 days.

[0009] The present invention also provides corresponding platelets, wherein butyric acid or a pharmaceutically acceptable salt thereof is added to the provided platelets. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 The preservation effect of each group on platelets in Example 1 of the present invention; wherein, Figure 1 .A is a flow cytometric analysis of the effect of the butyrate buffer PAS-Bu (Glu-) without glucose on the externalization of phosphatidylserine (PS), an apoptosis indicator of platelets during in vitro storage in Example 1 of the present invention; Figure 1 .B is the effect of PAS-Bu(Glu-) on PS eversion during platelet storage in vitro in Example 1 of the present invention (PS + Platelet percentage statistics); n = 3, all data in the histograms are mean ± SEM; *, p < 0.05; Figure 1 .C is a flow cytometric analysis diagram of PS eversion of platelets during in vitro storage by PAS-Bu in Example 2 of the present invention; Figure 1 .D is the effect of butyrate buffer (PAS-Bu) on the number of platelets during in vitro storage in Example 1 of the present invention; n=6, all data in the histogram are mean±SEM; NS: no statistical difference; *, p<0.05; Figure 1 .E is the effect of PAS-Bu on PS eversion during platelet storage in vitro in Example 1 of the present invention (PS + Platelet percentage statistics); n = 6, all data in the histograms are mean ± SEM; NS: no statistical difference; *, p < 0.05; **, p < 0.01.

[0011] Figure 2 The figure is a diagram showing the morphology and number of platelets in Example 2 of the present invention; Figure 2 .A is the effect of PAS-Bu on the morphology of preserved platelets in Example 2 of the present invention; Figure 2.B is the effect of PAS-Bu on the activation level of preserved platelets in Example 2 of the present invention (statistics of percentage of activated platelets); n=5, all data in the histogram are mean±SEM; NS: no statistical difference; *, p<0.05; **, p<0.01; ***, p<0.001; Figure 2 .C is the effect of PAS-Bu on the number of mitochondria in preserved platelets in Example 2 of the present invention (statistics of percentage of mitochondrial number); n=6, all data in the histogram are expressed as mean±SEM; NS: no statistical difference; *, p<0.05; **, p<0.01; ***, p<0.001.

[0012] Figure 3 The effect of platelet hemostasis function in Example 3 of the present invention; Figure 3 .A is the effect of PAS-Bu on platelet clot contraction ability in Example 3 of the present invention (analysis and statistics of clot contraction percentage); n=6, all data in the histogram are mean±SEM; NS: no statistical difference; *, p<0.05; **, p<0.01; Figure 3 .B is a schematic diagram of the effect of PAS-Bu on the platelet clot contraction ability in Example 3 of the present invention; Figure 3 .C is the effect of PAS-Bu on the in vivo hemostatic function after platelet transfusion in Example 3 of the present invention (statistics of mouse tail bleeding time); n=6, NS: no statistical difference; *, p<0.05; ****, p<0.001. DETAILED DESCRIPTION

[0013] Unless otherwise specified, the scientific and technical terms used herein are understood according to the knowledge of ordinary technicians in the relevant fields.

[0014] It is well known that platelets face a series of platelet storage lesions (PSLs) during the storage process, which ultimately result in a decrease in cell number, an increase in apoptosis and activation rates, morphological changes, mitochondrial dysfunction leading to a decrease in mitochondrial number, and impaired in vivo coagulation function after stored platelet transfusion.

[0015] Therefore, the present invention first analyzes the effect of PAS-Bu on the number of human platelets and finds that PAS-Bu can maintain the number of platelets; analyzes the effect of PAS-Bu on the PS eversion of human platelets and finds that PAS-Bu has the effect of reducing apoptosis of platelets during in vitro storage; further observes the effect of PAS-Bu on the morphology of human platelets and finds that PAS-Bu has the effect of maintaining the disc-shaped morphology of platelets and inhibiting the formation of platelet pseudopodia; and then analyzes the effect of PAS-Bu on the number of human platelet mitochondria and finds that PAS-Bu can maintain the number of platelet mitochondria stable and always at a high level; deeply analyzes the effect of PAS-Bu on the contraction ability of human platelet clots and finds that PAS-Bu can protect the function of blood clot contraction during in vitro storage. On this basis, the inventor further uses the SCID mouse platelet transfusion model to input PAS-Bu pretreated human platelets into SCID mice, and observes the bleeding time of the tail by cutting the tail, and finds that PAS-Bu can obviously maintain the hemostatic function of platelets.

[0016] The abbreviations used in this article have the following meanings:

[0017] PAS: PAS-III M buffer, the pH value of the buffer is 7.2-7.4, the buffer contains sodium chloride, potassium chloride, magnesium chloride, sodium citrate, sodium phosphate and sodium acetate; the concentration of each component can be configured according to actual needs. In the prior art, platelets are preserved using PAS-III M buffer and plasma mixture. The concentrations of each component in the PAS-III M buffer used in the following examples are 69.3 mmol / L sodium chloride, 5.0 mmol / L potassium chloride, 1.5 mmol / L magnesium chloride, 10.8 mmol / L sodium citrate, 26 mmol / L sodium phosphate, and 30 mmol / L sodium acetate; the pH value is 7.2-7.4.

[0018] PAS-Bu: It is composed of sodium butyrate, glucose, sodium chloride, potassium chloride, magnesium chloride, sodium citrate and sodium phosphate, that is, a solution in which sodium butyrate and glucose are dissolved in a PAS buffer solution without sodium acetate. In order to illustrate the preservation effect of the present invention on platelets relative to the existing PAS-III M buffer, the concentrations of sodium butyrate and glucose in the PAS-Bu used in the following examples are determined based on the concentration of sodium acetate in the PAS-III M buffer solution in the control group, the amount of plasma used in combination and the relevant buffering effect, and the concentrations of sodium butyrate and glucose in the experimental group are determined to be 5mmol / L and 7.5mmol / L, respectively. The PAS-Bu buffer used in the following examples contains 69.3mmol / L of sodium chloride, 5.0mmol / L of potassium chloride, 1.5mmol / L of magnesium chloride, 10.8mmol / L of sodium citrate, 26mmol / L of sodium phosphate, 5mmol / L of sodium butyrate, and 7.5mmol / L of glucose; the pH value is 7.2-7.4.

[0019] PAS-Bu (Glu-) buffer: It is composed of sodium butyrate, sodium chloride, potassium chloride, magnesium chloride, sodium citrate and sodium phosphate, that is, a solution in which sodium butyrate is dissolved in a PAS buffer without sodium acetate. In order to illustrate the preservation effect of the present invention on platelets relative to the existing PAS-III M buffer, the concentration of sodium butyrate in the PAS-Bu (Glu-) used in the following examples is determined based on the concentration of sodium acetate in the PAS-III M buffer in the control group and the relevant buffering effect. The PAS-Bu (Glu-) buffer used in the following examples contains 69.3 mmol / L sodium chloride, 5.0 mmol / L potassium chloride, 1.5 mmol / L magnesium chloride, 10.8 mmol / L sodium citrate, 26 mmol / L sodium phosphate, and 10 mmol / L sodium butyrate; the pH value is 7.2-7.4.

[0020] The present invention is further described below through examples. The platelets used in the examples were collected from healthy unpaid blood donations; sodium butyrate was purchased from Sigma, Annexin V-FITC Apoptosis Staining / Detection was purchased from BD; Mito-Tracker Green was purchased from MCE; the mice used were 20-25g SCID mice purchased from Shanghai South Model Organisms Technology Co., Ltd.; thrombin was purchased from Shanghai Yuanye Biotechnology Co., Ltd.; Anti-miceCD42b Antibody was purchased from Emfret.

[0021] Example 1: The inventors found that PAS-Bu can maintain the number of human platelets and alleviate apoptosis during in vitro storage

[0022] (1) Research plan

[0023] Healthy adults who had not taken any medication within 10 days were recruited as volunteers, and then apheresis platelets were prepared using a blood cell separator (Amicus 4R4580, Fenwal).

[0024] 5 mL of platelets were taken into two 15 mL centrifuge tubes, centrifuged at 2000 rpm for 15 min, and the precipitate (platelets) was retained. The upper plasma was collected and the plasma was centrifuged for a second time at 4000 rpm for 10 min. 5 mL of centrifuged plasma (control group / control group 1, PLASMA), 80% volume ratio of PAS-III M buffer + 20% volume ratio of plasma (control group 2, PAS-IIIM), PAS-Bu (Glu-) buffer (experimental group 1, PAS-Bu (Glu-)), PAS-Bu buffer (experimental group 2, PAS-Bu) were used to resuspend the above platelets, and the platelet number in each group was adjusted to 8 × 10 8 / mL;

[0025] Afterwards, each group was placed in a different platelet storage bag, and each group of platelets was placed in a platelet shaking storage box for in vitro storage (continuous shaking at 22±2°C).

[0026] (2) Platelet percentage test

[0027] On the 3rd, 5th and 7th days of storage, appropriate amount of platelets were collected from control group 1 and experimental group 1, and PS was labeled according to the Annexin V-FITCApoptosis Staining / Detection manual, and then the percentage of Annexin V positive platelets was detected by flow cytometry (BD FACSCanto). Figure 1 .As shown in A.

[0028] After testing, the percentage of Annexin V positive platelets was statistically analyzed as follows Figure 1 As shown in .B, the percentages of Annexin V positive platelets in the control group 1 on the 3rd, 5th and 7th days were 13.5%, 27.3% and 35.4%, respectively, which were lower than 22.5%, 39.6% and 58.0% in the experimental group 1 on the 3rd, 5th and 7th days, respectively; this result indicates that compared with the control group 1, although the proportion of platelet apoptosis is large in PAS-Bu(Glu-), the survival of a small number of platelets can be maintained.

[0029] 100 μL of platelets were collected from control group 1, control group 2 and experimental group 2 on the 0th, 3rd, 5th, 7th and 9th days of storage, and the platelet count was detected using a blood routine detector and the values ​​were recorded. The platelet counts on the 3rd, 5th, 7th and 9th days of each group were compared with the platelet counts on the 0th day of each group to calculate the percentages of the platelet counts on the 3rd, 5th, 7th and 9th days.

[0030] After testing, the percentage of platelets was statistically analyzed as follows Figure 1 As shown in .D, the percentage of platelet counts on days 3, 5, 7, and 9 in control group 1 were 95.5%, 92.8%, 91.3%, and 85%, respectively, which were not statistically different from those on days 3, 5, 7, and 9 in control group 2 (96.2%, 93%, 90.7%, and 87.2%);

[0031] The proportion of platelet count in experimental group 2 on the 3rd, 5th and 7th days was 95.3%, 92.3% and 91.3%, respectively, which was not statistically different from that in control group 1 and control group 2; but when the storage time was extended to the 9th day, the proportion of platelet count in experimental group 2 was 88.8%, which was significantly different from that in control group 1 (84.8%) and control group 2 (86.8%), and was significantly higher than that in control group 1 and control group 2.

[0032] In addition, the platelet percentages of experimental group 1 on days 3, 5 and 7 (22.5%, 39.6%, 58.0%) were significantly lower than those of experimental group 2 on days 3, 5 and 7 (95.3%, 92.3%, 91.3%); experimental group 2 was significantly higher than experimental group 1.

[0033] The above results indicate that PAS-Bu can still maintain the number of platelets at a high level as the storage time increases.

[0034] (3) PS-positive platelet detection

[0035] On the 3rd, 5th, 7th and 9th days of storage, appropriate amount of platelets were collected from control group 1, control group 2 and experimental group 2, and PS was labeled according to the Annexin V-FITC Apoptosis Staining / Detection manual, and then the percentage of Annexin V positive platelets was detected by flow cytometry (BD FACS Canto). Figure 1 .C.

[0036] After testing, the percentage of Annexin V positive platelets was statistically analyzed as follows Figure 1As shown in Figure E, the percentage of Annexin V positive platelets in control group 1 on days 3, 5, 7 and 9 were 7.4%, 10.9%, 15.6% and 29.4%, respectively, compared with 7.8%, 15.9%, 18.3% and 25.7% in control group 2 on days 3, 5, 7 and 9, there was no statistical difference in the apoptosis ratio on day 3, and the percentage of apoptosis in control group 2 on days 5 and 7 was significantly higher than that in control group 1, but on day 9, the apoptosis level in control group 2 tended to decrease compared with that in control group 1; this result indicates that compared with control group 1, control group 2 can reduce the percentage of Annexin V positive platelets on day 9;

[0037] The percentage of Annexin V positive platelets in experimental group 2 on days 3, 5, 7 and 9 were 6.5%, 9.5%, 11.7% and 21.9%, respectively, which was no different from that in control group 1 and control group 2 on day 3. However, on day 5, the percentage of Annexin V positive platelets in experimental group (9.5%) was lower than that in control group 2 (15.9%). As the storage time was extended to day 7 and beyond, the percentage of platelet apoptosis in experimental group 2 was significantly lower than that in control group 1 (15.6%, 29.4%) and control group 2 (18.3%, 25.7%), and there was a significant difference. The results showed that the use of PAS-Bu to store platelets would reduce the percentage of Annexin V positive platelets.

[0038] Annexin V is a fluorescent probe of PS, and PS externalization is a classic indicator of cell apoptosis. Therefore, the above results of this example confirm that PAS-Bu has the effect of alleviating apoptosis of human platelets during in vitro storage.

[0039] Example 2: The inventors found that PAS-Bu can maintain the disc-shaped morphology of platelets, inhibit their activation and maintain the number of platelet mitochondria

[0040] (1) Observation of platelet morphology

[0041] ① Take 50 μL (5×10 8 / mL), and added to 2% glutaraldehyde for fixation; ② Use different concentration gradients of ethanol to dehydrate the platelets of each group, and each concentration of ethanol is passed twice, each time for 15 minutes; ③ Wait for the sample to dry in order to completely remove the moisture in the sample to avoid the formation of water vapor in a vacuum environment; ④ After the sample is gold-sprayed, observe it under a scanning electron microscope.

[0042] The platelet morphology was observed to be Figure 2As shown in .A, the platelet morphology of experimental group 2 on days 3 and 5 was similar to that of control group 1 and control group 2. The platelets in this period were in a resting state and could maintain their own disc-shaped morphology. On day 7, compared with control group 1, control group 2 and experimental group 2, the platelet pseudopodia of control group 1 was the longest, followed by control group 2, and the least in experimental group 2. Moreover, the shape of platelets stored on day 9 in experimental group 2 was similar to that of platelets on day 5 in control group 1 and that of platelets on day 7 in control group 2. Once the platelets are stored in vitro for a longer time, their morphology will change, the cytoskeleton proteins will reorganize, the platelets will deform from the resting disc shape and extend pseudopodia to the surrounding areas, and the experimental group has significantly fewer cells forming pseudopodia than control groups 1 and 2. This result proves that adding a certain amount of PAS-Bu will maintain normal platelet morphology and inhibit the formation of platelet pseudopodia.

[0043] (2) Platelet activation assay

[0044] ① The platelets from control group 1, control group 2 and experimental group 2 were taken for scanning electron microscopy on the 3rd, 5th, 7th and 9th days of storage respectively; ② The number of activated platelets with pseudopodia was recorded, and the percentage of activated platelets on the 3rd, 5th, 7th and 9th days was calculated by comparing the number of activated platelets in each group on the 3rd, 5th, 7th and 9th days with the number of platelets in each group on the same day.

[0045] After calculation, the proportion of platelet activation is Figure 2 As shown in Figure B, the proportion of platelet activation in experimental group 2 on days 3, 5, 7 and 9 was 13%, 12.7%, 14.2% and 21%, respectively, which was significantly lower than 24.4%, 32.7%, 39% and 44% in control group 1; although there was no significant difference between the proportion of platelet activation in experimental group 2 (22.8% and 25.4% on days 3 and 5) and control group 2 (3 and 5), the proportion of platelet activation in experimental group 2 had a downward trend; on days 7 and 9, the proportion of platelet activation in experimental group 2 was significantly lower than 30.8% and 38% in control group 2. The results confirmed that PAS-Bu has the function of maintaining the disc-shaped morphology of human platelets and delaying the activation of human platelets during in vitro storage.

[0046] (3) Platelet mitochondrial number detection

[0047] ① The platelet counts of control group 1, control group 2 and experimental group 2 were adjusted to 1×10 on the 5th, 7th and 9th days using PBS. 9 / mL for later use; ② Take 5μL of the platelet suspension obtained in step ①, add mitochondrial Mito-Tracker Green live cell dye with a concentration of 200ng / mL to make up the volume to 100μL, mix thoroughly, and incubate at room temperature in the dark for 30min; ③ Add 400μL PBS to terminate the reaction, and detect on a flow cytometer (fluorescence setting is FITC).

[0048] After testing, the proportion of platelet mitochondria is Figure 2 As shown in Figure C, the proportion of mitochondrial numbers in experimental group 2 on days 5, 7, and 9 was 42.1%, 41.9%, and 39.1%, respectively, compared with 36.9%, 25.5%, and 21% on days 5, 7, and 9 in control group 1. The number of mitochondria in experimental group 2 was significantly higher than that in control group 1, and there was a statistical difference from day 7; the number of mitochondria in experimental group 2 on days 5, 7, and 9 was compared with 46.4%, 40.4%, and 38.7% on days 5, 7, and 9 in control group 2. Although the number of mitochondria in experimental group 2 was lower than that in control group 2 on day 5, when the time was extended to day 7 or even day 9, the number of mitochondria in experimental group 2 was significantly higher. This result proves that the addition of a certain amount of PAS-Bu will maintain the number of platelet mitochondria at a high level and prolong the in vitro storage time of platelets to day 9.

[0049] Example 3: The inventors found that PAS-Bu can protect the platelet clot contraction function during in vitro storage and promote the platelet hemostatic function in vivo.

[0050] (1) Platelet clot contraction function test

[0051] ① The platelet counts of control group 1, control group 2 and experimental group 2 were adjusted to 2.5×10 on days 3, 5, 7 and 9, respectively. 8 / mL, centrifuge at 2000rpm for 5min and discard the supernatant; ② resuspend the platelets of control group 1, control group 2 and experimental group 2 with fresh plasma of the same blood type and adjust the volume of each group to 500μL; ③ add thrombin (0.5U / mL) and CaCl2 (0.025mol / L) to each group in turn, incubate in a constant temperature water bath at 37℃ and start timing, observe (20min and 40min) the contraction of platelet clots and measure the size of blood clots at different times with Image J software.

[0052] Schematic diagram of platelet clot contraction Figure 3 .A, the clot shrinkage statistics are as follows Figure 3.B. After testing, the platelet clot ratio of experimental group 2 on the third day at 20 minutes was 67%, which was not significantly different from 79% of control group 1 and 75% of control group 2. At 40 minutes, the platelet clot ratio of experimental group 2 was 30%, which was statistically different from 50% of control group 1, but not different from 37% of control group 2. The platelet clot ratio of experimental group 2 on the fifth day at 20 minutes was 76%, which was statistically different from 94.5% of control group 1 and 90% of control group 2. Although the clots in the experimental group had a significant shrinking trend, there was no significant difference. At 40 minutes, the platelet clot ratio of experimental group 2 was 26%, which was statistically different from 52% of control group 1, but not different from 32% of control group 2. However, when the platelet storage time in vitro was extended to the seventh day, the platelet clot ratio of experimental group 2 The clot shrinkage size at 20min and 40min was 53.6% and 20.6%, both smaller than 67.3% and 29.3% of control group 1, and there was a significant statistical difference. The clot at 20min in experimental group 2 was significantly smaller than that at 20min in control group 2 (69%), but there was no significant difference at 40min. When platelets were stored in vitro for 9 days, the clot shrinkage size at 20min and 40min in the experimental group was 41% and 17.3%, both smaller than 57.8% and 19% of control group 1, and there was a significant statistical difference. There was no significant difference in the clot size at 20min (57.9%) and 40min (18.9%) between experimental group 2 and control group 2. The platelet clot shrinkage ratio is inversely proportional to its in vitro aggregation function. Therefore, the implementation results prove that PAS-Bu can protect the platelet clot shrinkage function during in vitro storage.

[0053] (2) In vivo hemostatic function testing of platelets

[0054] ① Knockout of platelets in SCID mice

[0055] Prepare SCID mice, and inject Anti-mice CD42b Antibody monoclonal antibody through the tail vein according to the instructions to clear the autologous platelets in the mice in advance.

[0056] ② Platelet transfusion

[0057] The preparation and treatment of platelets in this example were the same as in Example 1, except that the platelet suspension of control group 2 was reduced. Then, platelet suspension of control group 1 was taken on the 5th day of storage, and platelet suspension of experimental group 2 was taken on the 5th, 7th and 9th days of storage, with 1×10 9 Platelets were infused into the corresponding mice via the tail vein.

[0058] ③ Detection of mouse tail bleeding time

[0059] a. Stop timing 10 minutes after human platelets are transfused into SCID mice; b. Place the mouse horizontally on the workbench and fix it, cut its tail 2 mm, restart timing from zero, and place the mouse tail naturally vertically in a 1 mL EP tube; c. Wipe off a drop of blood with the wall of the EP tube every 30 seconds until bleeding stops, and finally record the bleeding time.

[0060] Statistical analysis of mouse tail bleeding time Figure 3 .C, the average hemostasis time of experimental group 2 on the 5th day was 148 seconds, which was significantly shorter than 404 seconds of control group 1 on the 5th day. The average bleeding time of experimental group 2 on the 7th day was 149 seconds, which was significantly shorter than 555 seconds of control group 1 on the 7th day. There was no difference in the tail bleeding time of mice in experimental group 2 on the 5th day compared with that on the 7th day, and both were shorter than that of control group 1 on the 5th day. The bleeding time of experimental group 2 on the 9th day was 464 seconds. Although the tail bleeding time of mice in experimental group 2 on the 9th day was longer than that of control group 1 on the 5th day, it was significantly shorter than that of control group 1 on the 7th day. The tail bleeding time of mice is inversely proportional to platelet function. Therefore, the results of this embodiment show that PAS-Bu can promote the hemostatic function of platelets. At the same time, it further confirms that PAS-Bu pretreatment can extend the shelf life of platelets to the 9th day.

Claims

1. Application of PAS-Bu solution for preparing platelet preservative; the PAS-Bu solution is prepared from sodium butyrate, glucose, sodium chloride, potassium chloride, magnesium chloride, sodium citrate and sodium phosphate.

2. The use according to claim 1, characterized in that: The pH value of the PAS-Bu solution is: 6.6-7.4。 3. The use according to claim 1, characterized in that: The concentration of sodium butyrate in the PAS-Bu solution is greater than or equal to 5 mmol / L.

4. The use according to claim 1, characterized in that: The concentrations of the components in the PAS-Bu solution are: sodium chloride 69.3 mmol / L, potassium chloride 5.0 mmol / L, magnesium chloride 1.5 mmol / L, sodium citrate 10.8 mmol / L, sodium phosphate 26 mmol / L, sodium butyrate 5 mmol / L, and glucose 7.5 mmol / L.

5. A platelet preservative, characterized in that: The platelet preservative is prepared from sodium butyrate, glucose, sodium chloride, potassium chloride, magnesium chloride, sodium citrate and sodium phosphate.

6. The platelet preservative according to claim 5, characterized in that The pH value of the platelet preservative is 6.6-7.

4.

7. A platelet preservative, characterized in that: The platelet preservative is prepared from sodium butyrate, sodium chloride, potassium chloride, magnesium chloride, sodium citrate and sodium phosphate.

8. A platelet storage method, characterized in that: Platelets are stored using the platelet preservative as claimed in claim 5.

9. The platelet storage method according to claim 8, characterized in that: Keep at room temperature for less than or equal to 9 days.

10. A platelet, characterized in that: The platelet preservative according to claim 5 is added to the platelets.

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