Study On The Quality Standard Of Tongfengning Capsule And Its Anti Gout Pharmacodynamics

This context researched Tongfengning Capsule from the qualitative and quantitative aspects by Thin-Layer Chromatography（TLC）,High Performance Liquid Chromatography（HPLC）and Ultraviolet Visible spectrophotometry（UV）to establish the quality standards of TFN.The safety of TFN was detected by acute toxicity test;The effects of TFN on reducing uric acid level,analgesia and anti-inflammatory,and repairing damaged kidney were studied by the model of hyperuricemia in mice,gouty arthritis model and gouty nephropathy model in rats.Providing the theoretical basis for the further application of TFN through the above experiments.1.Study on TFN quality standard LC 3000-C18 column,the moving phase was acetonitrile(0.08 moL·L^-1)and potassium dihydrogen phosphate（10:90）;the column temperature was 35℃;the detection wavelength was 326 nm.NaNO 2-A1（NO 3）3-NaOH colorimetric method was used to determine the content of total flavonoids in TFN.Checking the Sinapis semen,Lysimachiae herba,Plantations semen refer to the Chinese Pharmacopoeia/2015 Edition.Results displayed that the results of TCL qualitative identification were clear,and there was no interference in the negative control;the Sinapine thiocyanate had a good linear relationship in the range of 0.8⁴ g.The average recovery was 101.4%,RSD=1.98.2.Investigation on safety of TFN The median lethal dose（LD50）and maximal tolerance dose（MTD）of TFN were determined by intrastate administration in mice.The results showed that median lethal dose of TFN in mice was not measured,MTD=48000 mg·kg^-1.The experimental results showed that TFN was safe and reliable for oral administration in the commonly used clinical dose.3.Effect of TFN on hyperuricemia The hyperuricemia model of mice was induced by yeast extract combined with oxygen acid.The serum levels of UA,Cr and BUN,and the activities of XOD and ADA in the liver homogenization of mice were used as the detection indexes.The pathological changes of kidney were observed by HE staining.The results illustrated that TFN high dose group can significantly reduce hyperuricemia mouse UA,BUN and Cr,XOD and ADA activity（P<0.05）,improve the kidney glomerular atrophy in some degree,renal interstitial fibrosis,tubular dilation and other pathological changes.The results showed that TFN had a better therapeutic effect on hyperuricemia.4.Effect of TFN on acute gouty arthritis（1）An experimental model of acute gouty arthritis was established by intraperitoneal injection of glacial acetic acid in mice,and the latent period of writhing and the number of writhing in mice were observed.The results displayed that TFN can prolong the writhing latency（P<0.05,P<0.01）,and reduced writhing（P<0.05,P<0.01）.The high dose of TFN was the most significant（P<0.05,P<0.01）.It can be seen that TFN had a good analgesic effect in the treatment of acute gouty arthritis.（2）A rat model of acute gouty arthritis was established by injecting sodium urate（MSU）into the ankle joint of rats by puncture.After modeling,the degree of ankle swelling,the levels of white blood cells,lymphocytes,neutrophils,serum and ankle tissue homogenization NO levels,serum UA levels,serum levels of IL-1βand TNF-αin the soft tissue and synovial fluid around the ankle joint were detected.The results displayed that 0⁸ h TFN was significantly decreased in the high-dose rats ankle swelling degree（P<0.01,P<0.05）,especially at fourth hour（P<0.01）;high dose of TFN for white blood cells,Lymphocytes and neutrophils of rats in blood decreased（P<0.01,P<0.05）compared with the model group.The content of UA in serum was significantly lower than that in model group（P<0.01,P<0.05）,and the levels of NO in serum was increased（P<0.05）.In high dose of TFN group,the content of NO in soft tissue and synovial fluid and the levels of inflammatory factors IL-1βand TNF-αwere significantly decreased（P<0.01,P<0.05）.There was no significant difference in the above parameters between the TFN low and medium dose groups（P>0.05）.Thus TFN may achieve therapeutic purpose by inhibiting inflammatory factors.5.Effect of TFN on gouty nephropathy The models of gouty nephropathy in the mice were set up by using adenine combined with ethambutol.At the end of the experiment,the levels of UA,Cr,MCP-1,TNF-α,XOD and BUN in the serum of mice,as well as the activity of COX-2 in the kidney homogenization of mice were detected,and the pathological changes of kidney,spleen and thymus were observed by HE staining.The results showed that high dose of group serum UA,Cr,BUN,MCP-1,TNF-alpha,COX-2 and XOD were significantly decreased（P<0.01,P<0.05）.Observed by pathological pictures,the pathological changes of the mouse had a certain degree of improvement.