Application of gp96 protein to preparation of medicine for treating idiopathic thrombocytopenic purpura

A platelet and idiopathic technology, applied in the field of biomedicine, can solve the problems of high mortality and low morbidity, and achieve the effect of reducing spleen coefficient and increasing the number

Inactive Publication Date: 2017-08-08
FOSHAN HEAT SHOCK BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows scientists to use this molecule to make medicine more effective against certain diseases caused by abnormal clotting or blood coagulation processes associated with these disorders.

Problems solved by technology

This patented technical problem addressed in this patents relates to finding ways to prevent autism spectrum disease caused by defibrillusion purging due to certain drugs like THP when patients develop it without detectable signs during their first visit.

Method used

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  • Application of gp96 protein to preparation of medicine for treating idiopathic thrombocytopenic purpura
  • Application of gp96 protein to preparation of medicine for treating idiopathic thrombocytopenic purpura
  • Application of gp96 protein to preparation of medicine for treating idiopathic thrombocytopenic purpura

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Obtaining of heat shock protein gp96

[0044](1) Purification of gp96 protein from placental tissue

[0045] 1. Tissue homogenate

[0046] Homogenization buffer formula: add PMSF (phenylmethylsulfonyl fluoride, molecular formula C 7 h 7 FO 2 S) to a final concentration of 1 mM (prepared in a beaker on ice: PMSF decomposes in an aqueous solution within a few hours, and the solution cannot be left overnight).

[0047] Take the beaker and place it on ice, quickly cut the placental tissue into fragments with a diameter of about 1-2 mm with scissors in the beaker, and then add homogenization buffer 8 times the weight of the tissue. Stir the tissue pieces into a glass homogenizer, homogenize until the bottom tissue pieces disappear, and then homogenize up and down for more than 15 times. The homogenate was poured into a centrifuge tube, centrifuged at 50,000 g for 60 min, and the supernatant was added to 1 / 10 volume of 10×PBS (pH7.5, 200 mM NaCl) for ConA-Sepha...

Embodiment 2

[0102] Example 2 Determination of heat shock protein gp96 in activated regulatory T cells

[0103] 1. Immunization of mice in groups

[0104] 100 mice weighing 18-22 g were randomly divided into placenta gp96 treatment group 1, placenta gp96 treatment group 2, placenta gp96 treatment group 3, yeast gp96 treatment group 1, gp96 treatment group 2, yeast gp96 treatment group 3, and insect gp96 treatment group Treatment group 1, insect gp96 treatment group 2, insect gp96 treatment group 3 and control group (10 in each group) were processed as follows respectively:

[0105] Placenta gp96 treatment group 1: On the first day of the experiment, the gp96 solution extracted from human placenta tissue in Example 1 was injected intraperitoneally; on the eighth day of the experiment, the gp96 solution extracted from human placenta tissue in Example 1 was injected intraperitoneally again; The gp96 solution extracted from human placenta tissue in Example 1 was injected intraperitoneally aga...

Embodiment 3

[0130] Example 3 Application of gp96 in the treatment of immune thrombocytopenic purpura

[0131] 1. Analysis of platelet count

[0132] 1. Preparation of guinea pig antiplatelet antiserum (APS)

[0133] Take healthy BALB / C mice, take blood from the eye socket, rest for 1-2h, centrifuge at 700rpm / min for 10min, take the upper serum and centrifuge at 1500rpm / min for 15min, take the upper serum and centrifuge at 900rpm / min for 10min, take the upper serum and centrifuge at 3000rpm / min for 10min, Discard the serum, get the precipitated platelets at the bottom, add 1mL of 1% (mass volume fraction) ammonium oxalate, stand still for 5min, centrifuge at 3000rpm / min for 10min, discard the upper liquid, wash the platelets at the bottom three times with normal saline, count, and dilute with normal saline to a concentration of 1 x 10 9 -2×10 9 pc / L, respectively mixed with Freund's complete adjuvant and Freund's incomplete adjuvant at a ratio of 1:1 to form a Freund's complete adjuvant...

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Abstract

The invention provides application of gp96 protein to preparation of a medicine for treating idiopathic thrombocytopenic purpura. The gp96 protein is heat shock protein gp96. According to the application provided by the invention, the heat shock protein gp96 is used for preparing the medicine for treating the idiopathic thrombocytopenic purpura for the first time; an experiment proves that the heat shock protein gp96 has functions of inducing regulatory T cells production, reducing spleen coefficient and improving the quantity of blood platelets. Therefore, the heat shock protein gp96 has important application value for treating autoimmune thrombocytopenic purpura and/or alleviating symptoms of the autoimmune thrombocytopenic purpura.

Description

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Claims

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Application Information

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Owner FOSHAN HEAT SHOCK BIOTECH CO LTD
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