Effects of Lactobacillus plantarum on gut barrier function in experimental obstructive jaundice

Table 1 Sequences of oligonucleotide primers and conditions for real-time polymerase chain reaction

Gene target	Genbank number (mRNA)	Oligonucleotide1 (5'- to 3'-)	Annealing temperature (°C)	Product size (bp)
Occludin	NM-031329	F: GCTCAGGGAATATCCACCTATC R: TTCTCCAGCAACCAGCATC	60	344
ZO-1	NM-001106266	F: CCACAGACATCCAACCAGC R: AGCCCAAAGAACAGAAGACC	60	247
Claudin-1	NM-031699	F: GCCTCCAATGCCGTTCT R: TGCCTGCGTCCCTCTTG	60	317
Claudin-4	NM-001012022	F: GTTCCCGCCAGCAACTATG R: CCTTCAGCCCCGTATCCA	60	282
JAM-A	NM-053796	F: CCTCCATCCAAGCCGACA R: CAAAGACCAAATCCCCTGAC	60	211
prkC	NM-001105713	F: GATGGACGGGGTCACGA R: CGCTTGGCAGGGTGTTT	60	165
β-actin	NM-031144	F: CAGGTCATCACTATCGGCAAT R: GAGGTCTTTACGGATGTCAAC	60	144

¹Primers were designed based on sequences of rat-corresponding genes from the GenBank database. JAM-A: Junction adhesion molecule-A; ZO-1: Zonula occludens-1.

Table 2 Levels of endotoxin, total bilirubin, alanine transaminase and aspartate transaminase (mean ± SD)

	Group I	Group II	Group III	Group IV	Group V
Endotoxin (ng/mL)	0.58 ± 0.09	17.12 ± 1.09a	14.25 ± 0.68	3.05 ± 0.78c	1.91 ± 0.54
TBIL (μmol /L)	3.0 ± 1.63	153.83 ± 25.73a	132.0 ± 23.09	23.75 ± 5.42c	9.0 ± 1.87
ALT (U/L)	60.8 ± 5.69	543.83 ± 184.09a	218.38 ± 91.09	118.63 ± 19.72c	97.0 ± 10.37
AST (U/L)	130.9 ± 27.42	980.5 ± 663.25a	512.75 ± 156.76	437.88 ± 42.41c	271.0 ± 28.93

^aP < 0.05 vs Group III;

^cP < 0.05 vs Group V. Group I : Sham-operation; Group II: Bile duct ligation (BDL); Group III: BDL + L. plantarum; Group IV: BDL + internal biliary drainage (IBD); Group V: BDL + IBD + L. plantarum. TBIL: Total bilirubin; ALT: Alanine transaminase; AST: Aspartate transaminase.

Table 3 Levels of D-lactate, diamine oxidase, superoxide dismutase, malondialdehyde, glutathione and glutathione

	Group I	Group II	Group III	Group IV	Group V
D-lactate (mmol/L)	1.723 ± 0.106a	4.236 ± 0.050c	3.599 ± 0.181	3.152 ± 0.123e	2.800 ± 0.129
DAO (U/L)	2.829 ± 0.438a	18.925 ± 1.485c	12.928 ± 1.544	10.198 ± 0.584e	7.109 ± 0.590
SOD (U/mg protein)	65.002 ± 4.397a	26.782 ± 1.979c	35.396 ± 1.328	43.916 ± 1.720e	53.066 ± 3.203
MDA (nmol/mg protein)	0.408 ± 0.054a	1.253 ± 0.154c	0.914 ± 0.108	0.672 ± 0.054e	0.540 ± 0.029
GSH (μmol/L)	21.091 ± 0.452a	7.235 ± 0.479c	8.431 ± 0.537	10.504 ± 0.481e	19.082 ± 0.455
GSSG (μmol/L)	2.974 ± 0.260a	23.753 ± 2.895c	12.795 ± 1.360	4.944 ± 0.207e	3.537 ± 0.343

^aP < 0.05 vs Group II;

^cP < 0.05 vs Group III;

^eP < 0.05 vs Group V. Group I : Sham-operation; Group II: Bile duct ligation (BDL); Group III: BDL + L. plantarum; Group IV: BDL + internal biliary drainage ( IBD); Group V: BDL + IBD + L. plantarum. DAO: Diamine oxidase; SOD: Superoxide dismutase; MDA: Malondialdehyde; GSH: Glutathione; GSSG: Glutathione.

Table 4 Expression (mRNA) ratio (studied genes/β-actin) for tight junction and protein kinase C in terminal ileum tissues of each experimental group (mean ± SD)

Genes	Group I	Group II	Group III	Group IV	Group V
Occludin	2.5458 ± 0.2260	0.4881 ± 0.0426a	0.9792 ± 0.2066	1.4902 ± 0.0720c	1.8976 ± 0.1049
ZO-1	7.2420 ± 0.4025	0.9541 ± 0.1629a	1.4064 ± 0.1632	2.8843 ± 0.1641c	4.0727 ± 0.2059
Claudin-1	1.9751 ± 0.0615	0.0546 ± 0.0336a	0.4741 ± 0.0897	0.9092 ± 0.1295c	1.4793 ± 0.2119
Claudin-4	42.8680 ± 7.5291	0.3546 ± 0.0916a	5.3245 ± 1.1801	8.7719 ± 1.4659c	15.9592 ± 2.8815
JAM-A	3..3259 ± 0.3704	0.4712 ± 0.1107a	0.9456 ± 0.1101	1.6270 ± 0.2153c	2.1006 ± 0.1534
PKC	6.6958 ± 0.9349	1.7959 ± 0.2992a	2.8281 ± 0.3287	3.7178 ± 0.5110c	4.7235 ± 0.4958

^aP < 0.05 vs Group III;

^cP < 0.05 vs Group V. Group I: Sham-operation; Group II: Bile duct ligation (BDL); Group III: BDL + L. plantarum; Group IV: BDL + internal biliary drainage (IBD); Group V: BDL + IBD + L. plantarum. JAM-A: Junction adhesion molecule-A; PKC: Protein kinase C.