Studies On The Changes Of Alkaloid Content And Subacute Toxicity Before And After The Processing Of Aconitum Vilmorinianum KOM.

Aconitum,Aconitum carmichaelii,Aconitum kusnezoffii Reichb,etc.and other aconite plants which are typical representatives of medicinal toxic Chinese medicines are commonly used clinically as toxic Chinese medicines in China,with the earliest recorded toxic herbs in Chinese history.The main toxic components of these plants are aconitine alkaloids and their main target organs for toxic effects are the heart and nervous system.A large number of studies have shown that alkaloids contained in Aconitum are both toxic and medicinal components which were widely used in clinical treatment with many pharmacological action such as strengthen heart,analgesic,anti-inflammatory,anti-tumor,lower blood pressure,reduce vascular permeability and so on.Because of the severe toxicity of aconitine alkaloids and the effective therapeutic dose extremely closed to the toxic dose or lethal dose,aconitine alkaloids could cause poisoning or even death with improper processing or slightly inadvertent.Aconitum vilmorinianum Kom.is the dry root of the Aconitum vilmorinianum Komarov with the mild medicinal properties,spicy taste and toxicity.Besides,it had the effects of treating rheumatism,promoting blood circulation and relieving pain,mainly used for the treatment of wind-cold dampness,stroke,bruises and so on.The Aconitum vilmorinianum Kom.,an important part of the prescriptions for wound medicines such as Yunnan Red Medicine,Swelling and Anti-Aerosol,has been included in the 2005 edition of Yunnan Pharmaceutical Standards.The main active constituents and toxic components of Aconitum are aconitine alkaloids.Different from the most of the Aconitum plants recorded in the Pharmacopoeia,the main alkaloids contained in the Aconitum vilmorinianum Kom.were yunnaconitine and 8-deacetyl yunnaconitine for non-fermented products and processed products respectively.However,there was no the medicinal herbs quality standard of toxicity evaluation in the Pharmacopoeia.At present,the research on Aconitum vilmorinianum Kom.at home and abroad mainly focuses on the study of its chemical constituents,and its toxicity research only stays in the stage of acute toxicity research.Subacute toxicity studies and long-term toxicity studies have not been reported.The main purposes of this study are as follow :?1?to investigate the changes of the content of yunaconitine and 8-deacetyl yunaconitine before and after the preparation of Aconitum vilmorinianum Kom.?2?to explore the toxic target organs of Aconitum vilmorinianum Kom.and relationship between the content and toxicity of Aconitum vilmorinianum Kom.?3?By comparing the chemical composition changes before and after the processing,the damage degree of toxic target organs and the change of blood pressure,the attenuating effect of Aconitum vilmorinianum Kom.was discussed.The main research methods are as follows:1 To determine the content of yunaconitine and 8-deacetyl yunaconitine before and after the preparation of Aconitum vilmorinianum Kom.by ultra performance liquid chromatography;2 The Bliss method was used to determine the median lethal dose(LD₅₀)of the rats of the radix chinensis and the maximum dose of the rats of the Aconitum vilmorinianum Kom.3 Continuous intragastric administration for 14 days,observe the changes of II lead electrocardiogram in anesthetized rats;detect the changes in the level of alanine aminotransferase?ALT?,aspartate aminotransferase?AST?,blood urea nitrogen?BUN?,creatinine?CR?,creatine kinase?CK?,lactate dehydrogenase?LDH?in serum;observe thechanges in organ index of the heart,liver and kidney and pathological histomorphological changes;observe the changes in blood pressure systolic blood pressure,diastolic blood pressure and mean pulse pressure.The main results are as follows:1 The content of yunaconitine in Aconitum vilmorinianum Kom.was 263.96 ?g/g,and 8-deacetyl yunaconitine was not detected.The content of yunaconitine was not detected and 8-deacetyl yunaconitine was 568.47?g/g in the processed products.2 The LD₅₀ of Aconitum vilmorinianum Kom.was 4.2 g crude drug/kg,the toxicity of the processed products was too low to determine its LD₅₀,and the maximum dose of the processed products was 24.0 g crude drug/kg;3 After continuous intragastric administrated for 14 days,continuous observated for 60 min,anesthetized rats in the Aconitum vilmorinianum Kom.group had symptoms of arrhythmia such as ventricular premature beats,ventricular dichotomy and ventricular fibrillation,and bradycardia occurred in the early stage of the processed product group.After 60 min,it returned to normal.Compared with the blank group,the amplitudes of P wave and R wave of the three doses of Aconitum vilmorinianum Kom.were significantly increased?P<0.05?,and the amplitudes of T wave were significantly decreased?P<0.05?.There were no significant differences in Q wave amplitude,S wave amplitude,ST amplitude and ST time?P>0.05?.There was no significant difference in heart rate,QRS time,PR interval and QTc interval between the low dose group?P>0.05?.The heart rate of the middle dose and high dose group of Aconitum vilmorinianum Kom was significantly lower?P<0.05?,the QRS time of the high dose group was significantly longer?P<0.05?,and the QRS time of the middle dose group was significantly longer at 30 min and 45 min?P<0.05?.The QTc interval was significantly shorten in the high dose group?P<0.05?,and the QTc interval in the middle dose group was only significantly shorten at 30 min and 45 min?P<0.05?.There were no significant differences of the electrocardiogram of the rats in the same group at different time points in the indicators?P>0.05?.Compared with the blank group,there were no significant differences in the serum biochemical parameters of the processing group.The levels of ALT and AST were significantly increased?P<0.05?in the low-dose group and the levels of ALT,AST,CR,CK.LDH were significantly increased?P<0.05?in the middle dose group.The levels of ALT,AST,CR,BUN,CK and LDH in the high dose group were all significantly increased?P<0.05?.Compared with the control group,heart,kidney,liver changed in rats as follows:?1?the gap between the myocardial fibers of the raw group was significantly enlarged,and the infiltration of inflammatory cells was obvious.The myocardial fibers in the processed product group were arranged neatly,and individual inflammatory cells infiltrated in the interstitial.?2?The kidneys in the low-dose group of Aconitum vilmorinianum Kom.had normal morphological structure.The structure of the glomeruli and tubules in the processed product group was clear,the renal tubular epithelial cells were not degenerated and necrotic,the glomerular capsule was non-adhesive and narrow,and the interstitial had a small amount of inflammatory cell infiltration.In the medium-dose group and the high-dose group,the renal proximal tubular epithelial cells were edematous,the boundary was unclear,there was bleeding inside,and the interstitial cells infiltrated significantly.?3?In the processed product group,the cells on the hepatic cord were arranged neatly,a small amount of inflammatory cells infiltrated,and the cells in the low-dose hepatic cord of the raw group were disordered.The hepatocytes in the middle dose group were swollen,the nucleus was deeply stained,and the cells on the hepatic cord were disordered.The liver of the high dose group group showed fatty lesions,central venous congestion,hepatic sinus congestion,obvious inflammatory cell infiltration,disordered cell arrangement on the hepatic cord,the nucleus is deeply stained and the dinucleus is visible.Conclusion : Aconitum vilmorinianum Kom.has damage to heart,liver and kidney,especially to the heart and liver.The degree of organ damage is positively correlated with drug dosage.After processing,the content of toxic components and the damage to the toxic target organs were reduced simultaneously.Processing had attenuated effect on Aconitum vilmorinianum Kom.