284 216 4 1 41 4 [ ] (BCG+LPS) 6 NIH (1 5 25mg/kg) (15mg/kg) 1 ALT AST NO IL-1 TNF- Real-time PCR IL-6 inos mrna ALT AST (P<.1) NO IL-1 TNF- (P<.1) IL-6 inos mrna (P<.1) [ ] [ ] R378.992 [ ] A [ ] 577-742(216)4-284-5 [DOI] 1.11855/j.issn.577-742.216.4.5 Protective effect of mycelial polysaccharides from Cordyceps sinensis on immunological liver injury in mice DONG Kai-zhong 1, GAO Yong-sheng 2, WANG Xiao-heng 1, MA Yan-qing 1, SU Lu 1 1 Medical College, Northwest University for Nationalities, Lanzhou 733, China 2 Tianzhu Tibetan Autonomous County People s Hospital, Wuwei, Gansu 7332, China This work was supported by the Fundamental Research Funds for the Central Universities (ZYZ21292) [Abstract] Objective To explore the protective effects of mycelial polysaccharides from Cordyceps sinensis (MPCS) on BCG+LPS-induced liver injury in mice. Methods The immunological liver injury mice model was reproduced by giving bacillus Calmette-Guerin (BCG) and lipopolysaccharide (LPS). Sixty NIH mice were randomly assigned into 6 groups (1 each): normal control group, model group, mycelium polysaccharide in high (1mg/kg), medium (5mg/kg) and low (25mg/kg) dose group, and bifendate (15mg/kg) treatment group. The serum transaminase levels of alanine ALT and AST were assayed with ELISA, nitric oxide (NO) in serum was measured by nitrate reductase method, and the liver homogenate was prepared for the determination of the contents of interleukin-1 (IL-1 ) and tumor necrosis factor- (TNF- ). The mrna expression levels of IL-6 and inos in hepatic tissue were assessed using RT-PCR. Results In the mice of immunological liver injury, mycelial polysaccharides from Cordyceps sinensis obviously lowered the serum ALT and AST levels (P<.1), high dose MPCS significantly reduced the serum NO and liver tissue IL-1 and TNF- levels (P<.1). Compared with the model group, high and medium dose MPCS significantly reduced the expression levels of IL-6 and inos mrna in hepatic tissues (P<.1). Conclusion MPCS shows a certain protective effect on immunological liver injury induced by BCG plus LPS in mice. [Key words] mycelium; polysaccharides; liver injury [1] (Cordyceps sinensis) [ ] (ZYZ21292) [ ] [ ] 733 ( ) 7332 ( ) [2] [3] (mycelial polysaccharides from Cordyceps sinensis MPCS) (BCG+LPS)
Med J Chin PLA, Vol. 41, No. 4, April 1, 216 285 1 1.1 SPF NIH 2±2g SCXK( )211-4 (Sigma ) ( ) ALT AST NO ( ) IL-1 TNF- (RD ) ( ) 1.2 (BCG) (LPS) [4-5] BCG (.2ml/ 5 1 7 ) 12d LPS(.2ml/ 7.5 g) 1.3 NIH 6 6 1 (1mg/kg) (5mg/kg) (25mg/ kg) (15mg/kg) 1d 1 /d(.5% ) LPS( ) 16h 12h 1.4 1g [6] 1 12min 1:15 4 4 95% 4 24h 8r/min 1min 2 5 Sevag [7] DEAE-cellulose 1.5 ALT AST ALT AST ALT AST (U/L) 1.6 NO ( ) [8] 1min 55nm (A) NO 1.7 IL-1 TNF- 1% ELISA IL-1 TNF- 1.8 Real-time PCR IL-6 inos mrna Primer Premier 5. GenBank mrna IL-6 5'-CCACTTCACAAGTCGGAG- GCTTA-3' 5'-GCAAGTGCATCATCGTTGTTC- ATAC-3' inos 5'-GGCAGCCTGTGAGACCTT- TG-3' 5'-GCATTGGAAGTGAAGCGTTTC-3' -actin 5'-CCACAGCTGAGAGGGAAATC-3' 5'-TCTCCAGGGAGGAAGAGGAT-3' Total RNA RNA 37 1h 95 3min Real-time PCR 93 3min 93 3s 55 45s 72 45s 38 -actin IL-6 inos mrna 1.9 SPSS 13. x±s Student-t χ 2 P<.5 2 2.1 ALT AST (P<.1) ALT AST (P<.1 P<.5 1) 2.2 NO NO Transaminase concentration (U/L) 25 2 15 1 5 Median dose MP group ALT AST 1 BCG+LPS Fig.1 Effects of mycelial polysaccharides from Cordyceps sinensis on serum ALT and AST levels of mice with immunological liver injury induced by BCG plus LPS P<.1 compared with control group; P<.1, P<.5
286 216 4 1 41 4 (P<.1) NO (P<.1 P<.5 2) NO content ( mol/l) 25 2 15 1 5 Median dose MP group 2 BCG+LPS NO Fig.2 Effects of mycelial polysaccharides from Cordyceps sinensis on serum NO levels of mice with immunological liver injury induced by BCG plus LPS P<.1 compared with control group; P<.1, P<.5 2.3 IL-1 TNF- IL-1 TNF- (P<.1) IL-1 TNF- (P<.1 P<.5 3) 18 16 14 Median dose MP group 12 1 8 6 4 2 IL-1 TNF- 3 BCG+LPS IL-1 TNF- Fig.3 Effects of mycelial polysaccharides from Cordyceps sinensis on IL-1 and TNF- levels in liver homogenate of mice with immunological liver injury induced by BCG plus LPS P<.1 compared with control group; P<.1, P<.5 Content (pg/ml) 2.4 IL-6 inos mrna PCR IL-6 inos mrna (P<.1) IL-6 inos mrna (P<.1 P<.5 4) IL-6 inos mrna mrna expression 1..8.6.4.2 MediandoseMPgroup IL-6 4 IL-6 inos mrna Fig.4 Effects of mycelial polysaccharides from Cordyceps sinensis on the expression of IL-6 and inos mrna in liver tissues of mice with immunological liver injury P<.1 compared with control group; P<.1, P<.5 3 inos [9-11] BCG LPS [12] [13] [14] [15] BCG LPS BCG T LPS
Med J Chin PLA, Vol. 41, No. 4, April 1, 216 287 TNF- IL-1 NO [16] inos LPS Kupffer inos L- NO NO NO NO DNA [17] Kuffer TNF- TNF- NO IL-1 IL-2 IL-6 TNF- [18] IL-1 TNF- IFN- IL-6 IL-8 [19] IL-1 NO inos mrna [2] ALT AST NO 1mg/kg ALT AST NO NO TNF- IL-1 1mg/kg TNF- IL-1 BCG/LPS TNF- IL-1 NO [1] Wen Q, Dong RQ. Clinical pharmacology of the immune system[m]. Beijing: Chemical Industry Press, 21. 7-72. [,. [M]. :, 21. 7-72.] [2] DongKZ,FuSW,ShengL,et al. EffectofCordyceps sinensis mycelium on serum vasoactive intestinal peptide and substance P in mice with intestinal dysbacteriosis[ J]. Med J Chin PLA, 214, 39(11): 873-876. [,,,. VIP SP [J]., 214, 39(11): 873-876.] [3] Zhan XT, Lai GP, Zhu DX. Study progress in pharmacological effects of Cordyceps sinensis and its ferment mycelia[ J]. Med Recap, 212, 18(1): 1566-1569. [,,. [J]., 212, 18(1): 1566-1569.] [4] Kobayashi S, Nishihira J, Watanabe S, et al. Prevention of lethal acute hepatic failure by antimacrophage migration inhibitory factor antibody in mice treated with bacille Cahnette-Guerin and lipopolysaccharide[ J]. Hepatology, 1999, 29(6): 752-759. [5] Yoneyama H, Harada A, Imai T, et al. Pivotal role of TARC, a CC chemokine, in bacteria-induced fulminant hepatic failure in mice[ J]. Clin Invest, 1998, 12(11): 1933-1941. [6] Liu JH, Li FK, Jia DR, et al. Research on extraction, purification and physicochemical properties of mycelia polysaccharides of Hirsutella sinensis[ J]. Food Ferment Indust, 214, 4: 222-226. [,,,. [J]., 214, 4: 222-226.] [7] Yan JK, Li L, Wang ZM, et al. Structural elucidation of an exopolysaccharide from mycelial fermentation of a Tolypocladium sp.fungus isolated from wild Cordyceps sinensis[ J]. Carbohydrate Polymers, 21, 79: 125-13. [8] Wang YZ, Hu SS. A novel nitric oxide biosensor based on electropolymerization poly (toluidine blue) film electrode and its application to nitric oxide released in liver homogenate [ J]. Biosens Bioelectron, 26, 22: 1-17. [9] Zou Y, Yang Y, Li J, et al. Prevention of hepatic injury by a traditional Chinese formulation, BJ-JN, in mice treated with Bacille-Calmette-Guerin and lipopolysaccharide[ J]. J Ethnopharmacol, 26, 17: 442-448. [1] Ye XL, Liu Y, Chen RJ, et al. Analysis of evolution and virological characteristics of rti233v mutations in the hepatitis B virus reverse transcriptase domain[ J]. Med J Chin PLA, 215, 4: 188-193. [,,,. rti233v [J]., 215, 4: 188-193.] [11] Yao WM, Liu Y, Si LL, et al. Inhibitory effect of herbal concoction Suduxing on hepatitis B virus replication in vitro[j]. Med J Chin PLA, 214, 39: 121-124. [,,,. [J]., 214, 39: 121-124.] [12] Zhang AP, Shen YX, Wei W, et al. Protectiveeffectoftotal glucosides of Mudan cortexon immunological liver injury in mice[ J]. Chin Pharm Bull, 1997, 13(5): 435-437. [,,,. [J]., 1997, 13: 435-437.] [13] Wang SY, Shiao M. Pharmacological functions of Chinese medicinal fungus Cordyceps sinensis and related species[ J]. J Food Drug Anal, 2, 8(8): 248-257. [14] Chen JG, Lai WQ, Mei S, et al. Toxicological assessment of Cordyceps sinensis berkelysacc[ J]. Chin J Health Lab Tech, 29, 19: 417-419. [,,,. [J]., 29, 19: 417-419.] [15] Deng ZH, Lin XY, Li WM, et al. Protective effects of mycelium of cultured Cordyceps sinensis on test is in diabeticrats[ J]. Med J Chin PLA, 25, 3(4): 317-319. [,,,. [J]., 25, 3(4): 317-319.] [16] Guler R, Olleros ML, Vesin D, et al. Inhibition of inducible nitric oxide synthase protects against liver injury induced by mycobacterial infection and endotoxins[ J]. J Hepatol, 24, 41(5): 773-781. [17] Hu CM, Jiang H, Liu HF, et al. Effects of LJTF on immunological
288 216 4 1 41 4 liver injury in mice[ J]. Anhui Med Pharm J, 28, 12(4): 295-297. [,,,. [J]., 28, 12(4): 295-297.] [18] Nadler EP, Dickinson EC, Beer-Stolz D, et al. Scavenging nitric oxide reduces hepatocellular injury after endotoxin challenge[ J]. Am J Physiol Gastrointest Liver Physiol, 21, 281: G173-G181. [19] Dinarello CA. Interleukin-1 and interleukin-1 antagonism[ J]. Blood, 1991, 77(8): 1627-1652. [2] Nussler AK, Billiar TR. Inflammation, immunoregulation, and indncible nitric oxide synthase[ J]. J Leukoc Biol, 1993, 54: 171-178. ( 215-12-3 216-2-2) ( )