717解毒合剂调控巨噬细胞表型转化改善蝮蛇毒致局部组织损伤的作用及机制OA

OBJECTIVE To explore the effect and potential mechanism of 717 Jiedu decoction in ameliorating local tissue injury induced by Gloydius brevicaudus venom via regulating macrophage phenotype transformation.METHODS Rat blank-serum and drug-containing serum were prepared by intragastric administration of normal saline,tail-vein injection of anti-G.brevicaudus venom serum(0.6 mL per rat),and intragastric administration of 717 Jiedu decoction at doses of 0.495,0.99 and 1.98 g/kg(calculated as crude drug).M0-type macrophages derived from human leukemia monocytic THP-1 cells induced and differentiated by PMA were divided into normal control group,model group,positive control group,and low-,medium-and high-concentration 717 Jiedu decoction drug-containing serum groups.Except for the normal control group,all other groups were treated with 10 μg/mL G.brevicaudus venom working solution and 10%blank or drug-containing serum.After the cultivation period,the cellular ultrastructural changes in each group were observed;the proportions of M1-and M2-type macrophages,as well as the mRNA and protein expression of macrophage phenotype transformation and hypoxia-inducible factor-1α(HIF-1α)were detected.RESULTS Compared with the normal control group,cells in the model group presented ultrastructural injuries including mitochondrial swelling,cristae rupture and cytoplasmic vacuolization.The proportion of M1-type macrophages and the mRNA and protein expression levels of related phenotype transformation markers[inducible nitric oxide synthase,tumor necrosis factor-α,interleukin-6(IL-6),IL-1β]and HIF-1α were significantly increased(P＜0.05),whereas the proportion of M2-type macrophages and the mRNA and protein expression levels of related phenotype transformation markers(arginase-1,IL-10,mannose receptor C type 1 and chitinase-3-like protein 1)were significantly decreased(P＜0.05).Compared with the model group,cellular ultrastructural injuries were obviously alleviated,and the above-mentioned indexes were significantly reversed in the high-concentration 717 Jiedu decoction drug-containing serum group(P＜0.05).CONCLUSIONS 717 Jiedu decoction can exert anti-inflammatory and tissue-repairing effects by regulating the balance of M1-,M2-type macrophages phenotype transformation and repairing cellular ultrastructural damage.Its mechanism may be related to inhibiting the excessive activation of the HIF-1α pathway.