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
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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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