Triptolide

Spectra grade methanol and acetonitrile were purchased from Tedia (Fairfeild, USA). HPLC-grade formic acid was purchased from Kermel Chemical Reagent Co., Ltd. (Tianjin, China). Purified water was purchased from Wahaha (Zhejiang, Hangzhou, China). The reference standards triptolide, triptonide, triptophenolide, tripterifordin, wilforgine, wilforine, celastrol, demethylzeylasteral, wilforlide A, and pristimerin were purchased from Sichuan Weikeqi Biological Technology Co., Ltd. (Sichuan, China), and their purity was greater than 98%. Commercial Tripterygium glycosides tablets (TGT) were purchased from 13 factories (A-J), three batches of commercial Tripterygium wilfordii tablets (TWT) were from one factory (K), six different batches of commercial Tripterygium hypoglaucum hutch tablets (THHT) were purchased from two factories (L and M).

Hyp (purity>99%) and TP (purity>99%) were acquired via Sichuan Weikeqi Biological Technology Co., Ltd. TP was dissolved in DMSO and then diluted 20 times with normal saline for intraperitoneal injection.

Formic acid was HPLC grade obtained from Fisher (Fair Lawn, NJ, USA). Triptolide (Batch No. wkq18062706, purity ≥ 98%), triptonide (Batch No. wkq18030812, purity ≥ 98%), triptophenolide (Batch No. wkq18031301), wilforgine (Batch No. wkq18051009, purity ≥ 98%), wilforine (Batch No. wkq18080604, purity ≥ 98%), demethylzeylasteral (Batch No. wkq18031201, purity ≥ 98%), and celastrol (Batch No. wkq18042604, purity ≥ 98%) were purchased from Sichuan Weikeqi Biological Technology, Co., Ltd. (Sichuan, P.R. China).

The compounds isolated from Chinese traditional herbs for the anti-HIV-1 drug screen were purchased from Sichuan Weikeqi Biological Technology Co., Ltd., China. The purity of the compound was ≥ 98%, as was determined by high-performance liquid chromatography analysis. The compound samples were solved in dimethyl sulfoxide (DMSO) at a concentration of 50 mM.

DAPI 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium, MTT 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide and DMEM/F12 were obtained from Sigma (St. Louis, MO, USA). Aucubin (purity > 98%) and Triptolide (purity > 98%) were obtained from Weikeqi-Biotech Co., Ltd (Chengdu, China). The purity of Aucubin was detected using Waters alliance 2695-2998 HPLC.

Purified TP and EGCG were purchased from Sichuan Weikeqi Biological Technology Co., Ltd (Sichuan, China), their purity was verified with the HPLC method (purity

>98%) (Fig. 1). TP and EGCG were stored at 4C, prepared fresh prior to use, TP was

dissolved in a small amount of absolute ethyl alcohol, and EGCG was dissolved in a small

amount of DMSO

QE (purity > 98%) and TP (purity > 98%) were obtained from Sichuan Weikeqi Biological Technology Co., Ltd. (Sichuan, China). TLR4 protein antagonist Resatorvid (TAK-242) was purchased from Shanghai ZZBIO Co., Ltd. (Shanghai, China). Antibodies against TLR4, NF-κB, Foxp3 and β-actin were purchased from Wanleibio Co., Ltd.

Triptolide (HPLC purity 99.49%), triptophenolide (HPLC purity 99.29%), celastrol (HPLC purity 98.33%), wilforgine (HPLC purity 99.53%), wilforine (HPLC purity 99.32%) and wilfortrine (HPLC purity 99.09%) were supplied by Weikeqi Biological Technology Co., Ltd (Sichuan, China). Wilfordine (HPLC purity 98%) was purchased from ChemFaces Biochemical Co., Ltd (Wuhan, China) and irbesartan (HPLC purity 99.8%) was used as internal standard (IS) and was obtained from National Institute for Food and Drug Control (Beijing, China).

Triptolide was purchased from Sichuan Weikeqi-Biotech Company (Chengdu, China); Cycloheximide (CHX) was purchased from Dalian Meilun Biotechnology (Dalian, China); Probenecid was purchased from Shanghai Aladdin Bio-Chem Technology Co., Ltd. (Shanghai, China); A23187 was purchased from BioVision, Inc. (Milpitas, CA, USA), BAPTA-AM was purchased from Selleck Chemicals