Chloranthus multistachys Pei is a perennial herb and has been used as folk medicine to treat bone fracture,hemostasis swelling pain and pruritus cutanea for centuries in China.In this paper,the chemical constituents of C. multistachys and their anti-inflammatory activities were investigated.Twenty-two compounds were isolated by a variety of chromatographic techniques and identified as (7S,8R)-dihydrodehydrodiconiferyl alcohol (1),7-ketositosterol (2),6β-hydroxystigmast-4-en-3-one (3),6α-hydroxystigmast-4-en-3-one (4),4β,8β-dihydroxy-5α(H)-eudesm-7(11)-en-8,12-olide (5),13-epitorulosol (6),15-nor-14-oxolabda-8(17),12E-dien-19-oic acid (7),4α-hydroxy-8,12-epoxyeudesma-7,11-diene-1,6-dione (8),spicachlorantin A (9),fortunilide K (10),(1E,4Z)-8-hydroxy-6-oxogermacra-1(10),4,7(11)-trieno-12,8-1actone (11),henrilabdane B (12),2′-hydroxy-4,3′,4′,6′-tetramethoxychalcone (13),curcolonol (14),zederone epoxide (15),shizukaol B (16),cycloshizukaol A (17),henrilabdane A (18),12-(3-methyl-furan)-labd-8(17)-en-19-oic acid (19),ent-8(9)-pimarene-20-hydroxy-16-nor-15-oic acid (20),N-trans-feruloyltyramine (21) and grossamide (22).Compounds 2,4,21 and 22 were isolated from the genus Chloranthus for the first time,compounds 1-10 and 18-22 were isolated from C. multistachys for the first time.Additionally,compounds 5-17 were screened for anti-inflammatory activity by LPS-induced RAW264.7 cells.The results showed that compounds 5,10,11 and 13 exhibited moderate anti-inflammatory activity with the NO inhibition rates of 41.22%±8.28%,40.32%±15.14%,58.79%±8.16% and 43.85%±10.04%,respectively.