Evaluation of Long-Time Decoction-DetoxicatedHei-Shun-Pian(ProcessedAconitum carmichaeliDebeaux Lateral Root With Peel) for Its Acute Toxicity and Therapeutic Effect on Mono-Iodoacetate Induced Osteoarthritis

Background As a degenerative joint disease with severe cartilage destruction and pain, osteoarthritis (OA) has no satisfactory therapy to date. In traditional Chinese medicine (TCM),Aconitum carmichaeliDebeaux derivedHei-shun-pian(Hsp) has been developed for joint pain treatment. However, it causes adverse events in OA patients. Long-time decoction has been traditionally applied to reduce the aconite toxicity ofHspand other aconite herbs, but its detoxifying effect is uncertain. Methods Hspwas extracted with dilute decoction times (30, 60, and 120 min) and evaluated by toxicological, chemical, pharmacological assays. Acute toxicity assay and chemical analysis were employed to determine the toxicity and chemoprofile ofHspextracts, respectively. Since the detoxifiedHsp(dHsp) was defined, its therapeutic effect was evaluated by using an OA rat model induced by monosodium iodoacetate. dHspat 14 g/kg was orally administered for 28 days, and the pain assessments (mechanical withdrawal threshold and thermal withdrawal latency) and histopathological analyses (HE and safranin-O staining) were performed. Real-time PCR (qPCR) was applied to determine the molecular actions of dHspon cartilage tissue and on chondrocytes. MTT assay was conducted to evaluate the effect of dHspon the cell viability of chondrocytes. The cellular and molecular assays were also conducted to analyze the functions of chemical components in dHsp. Results The chemoprofile result showed that the contents of toxic alkaloids (aconitine, mesaconitine, and hypaconitine) were decreased but that of non-toxic alkaloids (benzoylaconitine, benzoylmesaconitine, and benzoylhypaconitine) were increased with increasing decoction time. Acute toxicity assay showed that onlyHspextract with 120 min decoction was non-toxic within the therapeutic dose range. Thus, it was defined as dHspfor further experiment. In OA experiment, dHspsignificantly attenuated joint pain and prevented articular degeneration from MIA attack. qPCR data showed that dHsprestored the abnormal expressions ofCol10,Mmp2,Sox5,Adamts4/5/9, and up-regulatedCol2expression in rat cartilage.In vitro, dHsp-containing serum significantly proliferated rat chondrocytes and regulated the gene expressions ofCol2,Mmp1,Adamts9, andAggrecanin a similar way as thein vivodata. Moreover, aconitine, mesaconitine, and hypaconitine exerted cytotoxic effects on chondrocytes, while benzoylaconitine and benzoylhypaconitine except benzoylmesaconitine exhibited similar