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Application of ultrasound microbubble-mediated siRNA interference GRK4 in targeted adjustment of natriuresis and blood pressure level

An ultrasonic microbubble and microbubble technology, applied in the field of biomedicine, can solve problems such as changing the level of urinary sodium excretion in the kidney, and achieve the effects of targeted prevention and treatment of hypertension, inhibition of expression, and stability of circulation in the body

Inactive Publication Date: 2016-02-03
THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there has been no report on the use of siRNA technology to interfere with GRK4 in the kidney, thereby changing the level of urinary sodium excretion in the kidney

Method used

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  • Application of ultrasound microbubble-mediated siRNA interference GRK4 in targeted adjustment of natriuresis and blood pressure level
  • Application of ultrasound microbubble-mediated siRNA interference GRK4 in targeted adjustment of natriuresis and blood pressure level
  • Application of ultrasound microbubble-mediated siRNA interference GRK4 in targeted adjustment of natriuresis and blood pressure level

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The experiment adopts lipid film microbubble "lipid fluoride" (the lipid film microbubble "lipid fluoride" is introduced in detail in the ZL02133720.9 patent), and its core gas is perfluoropropane, which is used for microbubble averaging when developing solid organs. The particle size is 2 μm, 98% of which are smaller than 8 μm, and the microbubble concentration is about 4×10 9 ~9×10 9 / ml, the particle size of the microbubbles can be significantly reduced after mechanical shaking at a faster speed.

[0030] Design 3 pairs of siRNA primers, the sequences of which are as follows:

[0031] Primer No. 1: 5'-ccuguauucuuagaccaaadtdt-3', SEQ ID NO.1;

[0032]Primer No. 2: 5'-ggcugucugauauaugaaadtdt-3', SEQ ID NO.2;

[0033] Primer No. 3: 5'-ggagagagcuccugaaguudtdt-3', SEQ ID NO.3.

[0034] The above-mentioned 3 pairs of primers were respectively wrapped with the above-mentioned ultrasonic microbubbles, and the steps of wrapping were as follows:

[0035] Since siRNA is ne...

Embodiment 2

[0044] Twelve SHR rats were randomly divided into the microbubble group and the “microbubble + GRK4siRNA (primer 1)” group, and the above-mentioned microbubbles loaded with GRK4siRNA were injected into the rats through the tail vein of the rats, and the ultrasound was started when the microbubbles were injected. , the ultrasonic conditions were the same as in Example 1, and the ultrasonic intervention was performed once every three days for 6 times for 20 days. In SHR rats treated with ultrasound-mediated microbubble disruption technology, the level of renal natriuresis was significantly changed. First of all, with the prolongation of the action time, compared with the control group, its 24-hour urine volume increased significantly (see appendix Figure 4 ); Secondly, the urinary sodium excretion rate (the urine sodium concentration measured by the electrolyte analyzer multiplied by the 24-hour urine output) also increased significantly with the continuous prolongation of the ...

Embodiment 3

[0046] Twelve SHR rats were randomly divided into the microbubble group and the “microbubble + GRK4siRNA (primer 1)” group, and the above-mentioned microbubbles loaded with GRK4siRNA were injected into the rats through the tail vein of the rats, and the ultrasound was started when the microbubbles were injected. , the ultrasonic conditions were the same as in Example 1, and the ultrasonic intervention was performed once every three days for 6 times for 20 days. In the SHR rats treated with ultrasound-mediated microbubble rupture technology, the mean arterial pressure was significantly lower than that of the control group (see appendix Figure 6 ).

[0047] The above experiments show that the present invention uses ultrasonic microbubble technology to carry siRNA to kidney-specific rupture, which can effectively inhibit the expression of GRK4 in the kidney, increase urinary sodium and urine excretion, and achieve the purpose of targeted prevention and treatment of hypertension....

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Abstract

The invention discloses application of ultrasound microbubble-mediated siRNA interference GRK4 in targeted adjustment of natriuresis and blood pressure level. A siRNA molecular sequence is 5minute-ccuguauucuuagaccaaadtdt-3minute. An ultrasound microbubble technology is adopted to carry siRNA to kidney specificity rupture, kidney GRK4 expression is effectively inhibited, urine sodium and urine volume excretion is increased, and the aims of targeted prevention and treating hypertension are achieved.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to the application of siRNA interference GRK4 mediated by ultrasonic microbubbles in targeted regulation of urinary sodium excretion and blood pressure levels. Background technique [0002] Hypertension and its complications are becoming the main reasons affecting people's health. Kidney is an important organ responsible for urinary sodium excretion, and it has received general attention in hypertension research. Among the many factors regulating blood pressure, G protein-coupled receptor kinase 4 (Gprotein-coupled receptor kinases 4, GRK4) has attracted special attention in recent years. First, GRK4 is located on chromosome 4pl6.3, which is considered to be closely related to the pathogenesis of human essential hypertension; and multiple known SNPs of GRK4 are also directly related to the pathogenesis of hypertension. Secondly, unlike other members of the GRK family, such as GR...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61K48/00A61K31/7088A61P9/12
Inventor 杨剑曾春雨周林李晓龙
Owner THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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