Research Article Antitumor Ability of Berberine Accompanied by Modulation of Gut Microbiome in Sarcoma-180 Tumor-bearing Mice

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2 OPEN ACCESS International Journal of Pharmacology ISSN DOI: /ijp Research Article Antitumor Ability of Berberine Accompanied by Modulation of Gut Microbiome in Sarcoma-18 Tumor-bearing Mice 1 Yanhong Yang, 2 Zili Lei, 2 Li Huang, 2 Fei Yang, 3 Na Zhang, 4 Jian Yuan, 3 Kundong Li, 3 Juan Chen and 3 Jufeng Zhang 1 The First Affiliated Hospital (School of Clinical Medicine), Guangdong Pharmaceutical University, Nong-Lin-Xia Road #19, Yue-Xiu District, 518 Guangzhou, People's Republic of China 2 Guangdong Research Center of Metabolic Diseases of Integrated Western and Chinese Medicine, Guangdong Pharmaceutical University, Guangzhou Higher Education Mega Centre, 516 Guangzhou, People's Republic of China 3 School of Life Science and Biopharmacology, Guangdong Pharmaceutical University, Guangzhou Higher Education Mega Centre, 516 Guangzhou, People's Republic of China 4 Department of Pathology and Guangdong Key Laboratory for Bioactive Drugs Research, Guangdong Pharmaceutical University, Guangzhou Higher Education Mega Centre, 516 Guangzhou, People's Republic of China 1st and 2nd author contributed equally to this paper

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5 12 (a) G/G1 G2M S-phase G/G1: 2.66% S: 68.26% 12 (b) G/G1: 19.16% S: 72.3% 9 9 Number 6 Number Channels (PL2APBS) Channels (PL2APBS) (c) G/G1: 17.99% S: 75.34% 12 (d) G/G1: 13.75% S: 76.6% Number 6 Number Channels (PL2APBS) Channels (PL2APBS) 1 (e) Cell production (%) Control 5 µmol LG 1 1 µmol LG 1 2 µmol LG 1 * * G1 Phase S G G G

6 1 7 (a) 1 7 (b) (c) 1 7 (d) Proportion of apoptic cells (%) (e) ** Control 5 µmol LG 1 ** ** 1 µmol LG 1 2 µmol LG 1 G G G

7 2 (a) 1.5 (b) Tumor indux * Spleen index 1..5 Control BBR. Control BBR 1.5 (c) Thymus index Control BBR (a) (b) (c) (d) Cell numbers of Ki-67 positive (e) BBR * Control

8 Relative abundance Taxonomy stack distribution of phylum Bacteriodetes Firmicutes Proteobacteria Tenericutes Cyanobacteria Deferribacteres Others Unclassified 25 BBR-1 BBR-2 BBR-3 BBR-4 BBR-5 BBR-6 Tumor-1 Tumor-2 Tumor-3 Tumor-4 Tumor-5 Tumor-6 Sample names Normal-1 Normal-2 Normal-3 Normal-4 Normal-5 Normal-6 Relative abundance Taxonomy stack distribution of family Bacteriodetes_S24-7_group Prevotellaceae Lachnospiraceae Ruminoco ccaceae Bacteroidaceae Rikenellaceae Porphyromonadacea Helicobacteraceae Alcaligenaceae Lactobacilaceae Others Unclassified 25 BBR-1 BBR-2 BBR-3 BBR-4 BBR-5 BBR-6 Tumor-1 Tumor-2 Tumor-3 Tumor-4 Tumor-5 Tumor-6 Normal-1 Normal-2 Normal-3 Normal-4 Normal-5 Normal-6 Sample names

9 PCO1 (18.16%) Normal-3 Normal-2 Normal-6 Normal-4 Normal-1 Normal-5 Tumor-4 BBR-4 BBR-6 BBR-3 BBR-1 BBR-5 BBR-2 Tumor-5 BBR Normal Tumor Tumor-1 Tumor-3 Tumor-2 Tumor-6 PCO2 (11.5%) Bacteriodaceae Acidaminococcaceae Rikenellaceae Anaeroplasmataceae Rhodospirillaceae Ruminococcaceae Spirochaetaceae Porphyromonadaceae Clostridiales_vadinBB6_group Streptococcaceae Bacteroidales_S24-7_group Alcaligenaceae Lactobacillaceae Coriobacteriaceae Erysipelotrichaceae Bifidobacteriaceae Helicobacteraceae Deferribacteraceae Lachnosporaceae Desulfovibrionaceae Peptococcaceae Verrucomicrobiaceae Clostridiaceae_1 Prevotellaceae Peptostreptococcaceae Normal-1 Normal-2 Normal-3 Normal-4 Normal-5 Normal-6 Tumor-1 Tumor-2 Tumor-3 Tumor-4 Tumor-5 Tumor-6 BBR-1 BBR-2 BBR-3 RRB-4 BBR-5 BBR-6 Sample names

10 ABC transporters Two-component system Purine metabolism Aminoacyl-tRNA biosynthesis Pyrimidine metabolism Starch and sucrose metabolism Amino sugar and nucleotide sugar metabolism Ribosome Peptidoglycan biosynthesis Porphyrin and chlorophyll metabolism Arginine and proline metabolism Homologous recombination Fructose and mannose metabolism Cell cycle-caulobacter Mismatch repair Nucleotide excision repair Oxidative phosphorylation RNA degradation Alanine, aspartate and glutamate metabolism Methane metabolism BBR-1 BBR-2 BBR-3 RRB-4 BBR-5 BBR-6 Tumor-1 Tumor-2 Tumor-3 Tumor-4 Tumor-5 Sample names Tumor-6 Normal-1 Normal-2 Normal-3 Normal-4 Normal-5 Normal-6

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