Preparation Of Huanglian San And Its Pro-tective Effect On Colistin-induced Kidney Injury In Mice

Colistin is a polypeptide antibiotic that has strong antibacterial activity against Gram-negative bacteria and is not prone to bacterial resistance.However,the clinical application of colistin is limited due to potential toxic side effects.In recent years,research on the reduction of toxicity of colistin has become a hotspot.Studies have found that traditional Chinese medicine/natural medicine is used as a protective agent in combination with colistin to alleviate its side effects.Rhizoma Coptis is a traditional Chinese medicinal material in my country.It is a plant of the genus Coptidis in the family Ranunculaceae.It has the effects of purging fire and detoxification,and has a synergistic effect when used in combination with antibiotics.The purpose of this study is to develop Huanglian San and investigate its effect on the nephrotoxicity of colistin,in order to provide a new idea for the combination of colistin in veterinary medicine,attenuation of toxicity and enhancement of efficacy.Firstly,an HPLC method for the determination of berberine in Rhizoma Coptis was established,and the content of 10 batches of Rhizoma Coptis from different origins and sources was determined.Secondly,the process of alcohol extraction of berberine in Rhizoma Coptis was optimized through single factor test and response surface test,and the optimal extraction process was finally determined as follows:ultrasonic time,ultrasonic power,ethanol concentration,extraction times,extraction temperature,and solid-liquid ratio respectively The settings were:28 min,70%,62%,3 times,50°C,1:12.Under this process,the transfer rate of berberine in Rhizoma Coptis was 63.21±1.06%.Subsequently,single factor test and orthogonal test were used to optimize the purification process of Rhizoma Coptis extract by acid precipitation method,and the optimal purification process was determined as follows:Ca（OH）₂neutralization condition p H=8.00,HCl acid precipitation condition p H=2.00,Na Cl salting-out concentration The purity of berberine was 65.94±1.15%after optimizing the process.Then,using anhydrous glucose as supplementary material,and Rhizoma Coptis extract:anhydrous glucose=1:3,mixed with equal volume increase method to prepare Huang Lian San,and carried out quality inspection and preliminary quality standard research on Huang Lian San.The test results show that the powder is a brown-yellow powder with uniform color and no mottled spots.It all passes through the No.5 sieve,the water content is1.87%,and the berberine content is 14.53±0.18%.Then conduct a preliminary stability study on Huang Lian San.It is suggested that the powder should be sealed and stored in a cool and dark place.Finally,a mouse kidney injury model was established with mucormycin and Huang Lian San was given orally to investigate the alleviating effect of Huang Lian San on the nephrotoxicity of mucormycin.Compared with the model group,the serum BUN and CRE in the Huang Lian San group were significantly reduced（P<0.05 or P<0.01）,the kidney tissues of mice were less damaged,the necrosis of renal tubular epithelial cells was reduced,the antioxidant capacity of the body was improved and the level of lipid peroxidation was reduced,indicating that the bulk could alleviate the nephrotoxicity caused by mucormycin.In conclusion,this study used Huang Lian as the original herb and optimised its extraction and purification process to produce a high purity extract of Huang Lian,supplemented with anhydrous glucose to produce Huang Lian San,which has a palliative effect on mucormycin-induced kidney injury.