By vantal Preliminary study on the method of establishing rat model of cardiorenal syndrome by nephrectomy combined with ISOin other words suture kit It is possible to develop in a good direction, and there are still many places worth looking forward to in the future. https://pinnaclemedics.com/
With the development of medical technology, the survival rate of patients with heart failure and kidney disease has been continuously improved, and the close relationship between heart and kidney diseases has been gradually discovered. Clinical treatment of cardiorenal syndrome is limited, and diuresis and ultrafiltration are the main treatment measures. In order to explore the mechanism of traditional Chinese medicine in treating cardiorenal co-injury, the experimental model of cardiorenal syndrome is indispensable.
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At present, the common modeling methods can be divided into surgical method and drug injection method. “5/6 nephrectomy to prepare rat model of cardiorenal syndrome” is one of the classic methods to prepare rat model of cardiorenal syndrome. Heart failure caused by renal failure is also one of the pathogenesis of cardiorenal syndrome, but it often takes a long time to prepare rat model of cardiorenal syndrome by a single method. After consulting the relevant literature, the research group finally chose isoproterenol (ISO) as the research direction. ISO is a non-selective β -receptor agonist, which can significantly accelerate the heart rate and improve the myocardial contractility. If it is used in large doses for a long time, it can directly induce myocardial injury.
Therefore, the research group chose to use ISO in low dose and short time after nephrectomy, in order to shorten the natural course of heart failure caused by renal failure and not directly induce myocardial injury. This method can simulate the disease process of cardiorenal syndrome to a high degree.
1 materials and methods
1.1 Main experimental materials
1.1.1 Animal condition Twenty SD male rats of clean grade, weighing (150 10) g, were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd..
1.1.2 Consumables and instruments ISO(SIGMA), pentobarbital sodium, penicillin sodium for injection, syringes (1 ml, 5 ml, 20mL), water for injection, iodophor, gelatin sponge, sterile gauze, capillary blood collection tube, coagulation-promoting centrifugal tube, pipette (100~1000μL), 10% neutral formaldehyde solution, etc. Sunrise automatic microplate reader, HH-4 constant temperature water bath pot, RT-3000 automatic plate washer, Eppendorf5427R desktop high-speed freezing centrifuge, etc.
1.1.3 detection reagents: rat urea nitrogen (BUN) enzyme-linked immunosorbent assay (ELISA) detection kit, rat brain natriuretic peptide (BNP)ELISA detection kit, and rat serum creatinine (Cr)ELISA detection kit.
1.2 experimental methods
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1.2.1 Grouping and modeling methods Twenty male SD rats were randomly divided into two groups, 9 rats in the normal group and 11 rats in the operation group. Rats in the normal group were not treated, and all rats in the operation group underwent 5/6 nephrectomy according to the “two-operation method”. After one week of operation, isoproterenol (2.5 mg/kg and 1.5mg/kg respectively) was injected subcutaneously for two days (24 hours apart) to establish the cardiorenal syndrome model.
1) One-stage operation (2/3 nephrectomy): After weighing and recording the weight of rats, 2% pentobarbital sodium 35mg/kg was injected intraperitoneally for anesthesia. After successful anesthesia, the rats were fixed in prone position on the rat operating table, and slowly pushed to the kidney from the left abdomen to determine the location of the operating area, and the skin of the operating area was prepared and disinfected with iodophor. A long incision of about 2cm was made at a distance of 1.5cm from the left ridge angle, and the kidney was pushed from the ventral side to the dorsal side again, so that the left kidney field of vision was fully exposed.
Bluntly separate the fat layer around the left kidney with tweezers, peel off the renal capsule, adrenal gland and renal pedicle, fully free the renal pedicle and clamp it with vascular forceps, then cut off part of the renal tissue from the upper and lower poles of the left kidney with curved scissors, and cut off 2/3 of the left kidney in total. Then use gelatin sponge to compress the incisions on both sides for 15~20s, loosen the vascular forceps and use sterile gauze to stop bleeding. When there is no fresh bleeding, return the remaining kidney and surrounding tissues to the body, and the gelatin sponge can be absorbed by itself without taking out. Inject 80,000 units of penicillin sodium into abdominal cavity before suture. Suture the muscle layer and skin layer by layer from the inside out, clean the incision and operation area with 0.9% sodium chloride injection and iodophor successively. After the operation, the rats were raised in a single cage and their vital signs were observed, and then put into the feeding cage for mixed culture after they fully recovered.