Lactobacillus rhamnosus LRa05 on glycemic control and gut microbiota in patients with type 2 diabetes

Table 1 Demographic and clinical characteristics of study participants

Variables	Placebo group (n = 38)	Intervention group (n = 38)	P value
Age (years)	54.68 (37.00-70.00)	59.55 (35.00-75.00)	0.054
Gender			0.365
Male	23 (60.5)	19 (50.0)
Female	15 (39.5)	19 (50.0)
BMI, kg/m2	25.35 (2.9)	25.61 (2.9)	0.692
Weight, kg	78.10 (58.50-105.00)	72.32 (55.00-102.50)	0.296
Smoking status	12 (31.6)	5 (13.2)	0.054
Drink Wine	1 (2.6)	6 (15.8)	0.107
Hypertension	14 (36.8)	17 (44.7)	0.483
Coronary heart disease	3 (7.9)	6 (15.7)	0.286
Diabetes duration (years)	7.18 (1.00-20.00)	10.16 (1.00-24.00)	0.025
Metformin use	32 (84.0)	31 (84.0)	0.956
Sulfonylureas use	12 (31.0)	15 (39.0)	0.156
Insulin use	4 (10.5)	5 (13.0)	0.256
Self-reported physical activity, MET (hours/week)	46.71 ± 6.3	50.07 ± 6.1	0.332

Data are presented as n (%) or median (25^th-75^th percentiles. BMI: Body mass index; MET: Metabolic equivalent of task.

Table 2 Blood indices and inflammatory markers over time

Index	Placebo group (n = 38)		Intervention group (n = 38)		Time (P value)1	Time group (P value)2	T2 vs T0 (P value)3	T3 vs T0 (P value)4
	mean ± SD	Median (Q1-Q3)	mean ± SD	Median (Q1-Q3)
WBC
Baseline	6.86 ± 2.04	6.39 (5.41-8.12)	6.52 ± 1.44	6.42 (5.66-7.02)	0.767	0.840	0.810	0.762
T2	6.71 ± 1.69	6.46 (5.43-7.99)	6.44 ± 1.76	6.33 (5.01-7.29)
T3	6.85 ± 1.87	6.51 (5.69-8.18)	6.41 ± 1.53	6.34 (5.07-7.62)
NEUT
Baseline	3.87 ± 1.42	3.40 (2.99-4.88)	3.73 ± 1.18	3.51 (3.13-4.11)	0.846	0.759	0.848	0.434
T2	3.83 ± 1.16	3.65 (2.89-4.36)	3.63 ± 1.46	3.59 (2.81-4.03)
T3	3.93 ± 1.22	3.63 (3.28-4.54)	3.60 ± 1.09	3.59 (2.70-4.25)
LYMPH
Baseline	2.29 ± 0.76	2.30 (1.85-2.54)	2.09 ± 0.60	2.11 (1.60-2.44)	0.537	0.537	0.399	0.288
T2	2.20 ± 0.81	2.11 (1.61-2.53)	2.08 ± 0.7	2.03 (1.61-2.52)
T3	2.20 ± 0.73	2.14 (1.77-2.43)	2.10 ± 0.72	2.06 (1.49-2.40)
MONO
Baseline	0.48 ± 0.16	0.46 (0.37-0.55]	0.48 ± 0.15	0.48 (0.37-0.57)	0.249	0.471	0.696	0.463
T2	0.46 ± 0.12	0.47 (0.36-0.53)	0.46 ± 0.15	0.45 (0.35-0.54)
T3	0.49 ± 0.13	0.47 (0.39-0.57)	0.46 ± 0.13	0.46 (0.39-0.59)
Eosinophil
Baseline	0.15 ± 0.13	0.13 (0.08-0.17)	0.17 ± 0.15	0.14 (0.09-0.21)	0.120	0.691	0.350	0.607
T2	0.15 ± 0.10	0.12 (0.08-0.20)	0.19 ± 0.13	0.18 (0.08-0.24)
T3	0.17 ± 0.12	0.13 (0.09-0.22)	0.20 ± 0.19	0.15 (0.10-0.23)
BASS
Baseline	0.05 ± 0.06	0.04 (0.03-0.06)	0.03 ± 0.01	0.03 (0.02-0.04)	0.413	0.253	0.224	0.258
T2	0.04 ± 0.02	0.04 (0.02-0.05)	0.04 ± 0.01	0.03 (0.03-0.04)
T3	0.04 ± 0.02	0.04 (0.03-0.06)	0.04 ± 0.02	0.03 (0.02-0.05)
RBC
Baseline	5.09 ± 0.38	5.11 (4.90-5.35)	5.09 ± 0.54	5.17 (4.86-5.45)	< 0.0001	0.140	0.145	0.763
T2	4.97 ± 0.40	5.01 (4.71-5.26)	5.05 ± 0.56	5.03 (4.76-5.39)
T3	4.96 ± 0.39	4.92 (4.76-5.25)	4.96 ± 0.54	5.04 (4.70-5.34)
HGB
Baseline	155.47 ± 11.93	155.50 (147.00-164.00)	150.47 ± 15.98	152.00 (139.00-162.00)	0.009	0.229	0.152	0.986
T2	152.92 ± 13.12	153.50 (142.00-160.00)	150.37 ± 15.67	150.00 (139.00-164.00)
T3	152.97 ± 11.67	152.00 (145.00-161.00)	147.95 ± 15.78	150.00 (138.00-158.00)
PLT
Baseline	226.68 ± 74.70	229.50 (168.00-269.00)	225.55 ± 53.94	221.00 (197.00-253.00)	0.983	0.658	0.634	0.360
T2	224.97 ± 73.63	229.50 (169.00-267.00)	226.34 ± 46.32	220.00 (202.00-250.00)
T3	223.58 ± 68.32	234.00 (179.00-260.00)	227.79 ± 52.87	218.00 (199.00-240.00)
SII
Baseline	401.27 ± 177.04	356.88 (305.20-524.20)	453.75 ± 301.90	379.10 (296.10-500.40)	0.866	0.766	0.642	0.4350
T2	424.75 ± 206.19	413.00 (237.20-519.03)	451.33 ± 346.54	385.80 (288.10-460.60)
T3	418.58 ± 174.58	405.50 (295.00-528.50)	443.45 ± 319.82	367.70 (286.60-521.70)
NLR
Baseline	1.77 ± 0.61	1.67 (1.35-2.23)	1.94 ± 0.87	1.64 (1.40-2.25)	0.773	0.735	0.622	0.378
T2	1.90 ± 0.75	1.81 (1.31-2.46)	1.95 ± 1.34	1.74 (1.29-2.11)
T3	1.88 ± 0.58	1.86 (1.47-2.32)	1.91 ± 1.00	1.69 (1.42-2.22)
PLR
Baseline	102.87 ± 32.01	106.70 (84.06-121.73)	117.05 ± 42.28	111.15 (91.70-144.44)	0.245	0.939	0.734	0.960
T2	108.27 ± 35.13	110.00 (77.70-141.90)	120.48 ± 42.92	114.70 (93.30-145.20)
T3	106.75 ± 33.37	108.60 (90.30-127.20)	120.6 ± 48.47	113.50 (85.00-139.80)
MLR
Baseline	0.22 ± 0.08	0.20 (0.17-0.26)	0.25 ± 0.11	0.22 (0.18-0.28)	0.977	0.314	0.869	0.132
T2	0.22 ± 0.07	0.21 (0.17-0.26)	0.25 ± 0.13	0.22 (0.17-0.29)
T3	0.23 ± 0.07	0.22 (0.18-0.28)	0.24 ± 0.09	0.23 (0.18-0.28)
SIRI
Baseline	0.88 ± 0.51	0.79 (0.62-1.01)	0.99 ± 0.78	0.84 (0.55-1.06)
T2	0.88 ± 0.46	0.81 (0.56-1.13)	0.99 ± 1.04	0.74 (0.48-1.06)	0.865	0.494	0.979	0.189
T3	0.92 ± 0.44	0.83 (0.65-0.95)	0.89 ± 0.54	0.72 (0.55-1.08)
NPR
Baseline	0.02 ± 0.01	0.02 (0.01-0.02)	0.02 ± 0	0.02 (0.01-0.02)	0.844	0.737	0.724	0.434
T2	0.02 ± 0.01	0.02 (0.01-0.02)	0.02 ± 0.01	0.02 (0.01-0.02)
T3	0.02 ± 0.01	0.02 (0.01-0.02)	0.02 ± 0.01	0.02 (0.01-0.02)
PAR
Baseline	5.00 ± 1.63	5.24 (3.38-6.09)	5.10 ± 1.28	4.98 (4.30-5.75)	0.299	0.523	0.897	0.365
T2	5.06 ± 1.63	5.02 (3.70-6.44)	5.15 ± 1.10	5.09 (4.52-5.63)
T3	5.05 ± 1.59	5.20 (3.76-6.21)	5.27 ± 1.41	5.01 (4.52-5.56)
CAR
Baseline	0.09 ± 0.05	0.06 (0.05-0.11)	0.08 ± 0.08	0.06 (0.04-0.09)	0.662	0.273	0.272	0.699
T2	0.11 ± 0.19	0.07 (0.05-0.12)	0.08 ± 0.04	0.07 (0.05-0.09)
T3	0.09 ± 0.06	0.07 (0.04-0.10)	0.09 ± 0.10	0.06 (0.04-0.10)
CLR
Baseline	1.76 ± 1.02	1.47 (0.91-2.19)	2.05 ± 2.18	1.13 (0.84-2.43)	0.717	0.273	0.254	0.725
T2	2.44 ± 4.03	1.46 (1.01-2.57)	1.87 ± 1.48	1.35 (1.00-2.31)
T3	1.79 ± 1.28	1.52 (0.98-2.22)	2.26 ± 2.98	1.26 (0.89-2.07)
TyG
Baseline	1.95 ± 0.74	1.84 (1.56-2.27)	1.90 ± 0.64	1.79 (1.41-2.29)	0.834	0.374	0.971	0.232
T2	1.98 ± 0.83	2.00 (1.44-2.45)	1.93 ± 0.57	1.96 (1.51-2.25)
T3	2.03 ± 0.85	1.93 (1.39-2.62)	1.83 ± 0.61	1.80 (1.47-2.19)

¹Time (P value): Assesses changes over time within each group.

²Time group (P value): Evaluates the interaction effect between time and group.

³T2 vs baseline (P value): This P value compares the outcome at time point T2 against the baseline.

⁴T3 vs baseline (P value): This P value compares the outcome at time point T3 against the baseline. NEUT: Neutrophils; LYMPH: Lymphocytes; MONO: Monocytes; HGB: Hemoglobin; PLT: Platelets; SII: Systemic inflammation index; NLR: Neutrophil-to-lymphocyte ratio; PLR: Platelet-to-lymphocyte ratio; MLR: Monocyte-to-lymphocyte ratio; SIRI: Systemic inflammation response index; NPR: Neutrophil-to-platelet ratio; PAR: Platelet aggregation ratio; CAR: C-reactive protein to albumin ratio; CLR: C-reactive protein to lymphocyte ratio; TyG: Triglyceride-glucose index; T0: Baseline; T2: Second time point of measurement (after 6 weeks); T3: Third time point of measurement (after 12 weeks); Q1: First quartile; Q3: Third quartile.

Table 3 Biochemical parameters, kidney function, and liver enzyme markers across the study duration

Index	Placebo group (n = 38)		Intervention group (n = 38)		Placebo group (n = 38)	Intervention group (n = 38)	T2 vs T0 (P value)	T3 vs T0 (P value)
	mean ± SD	Median (Q1-Q3)	mean ± SD	Median (Q1-Q3)
UREA
Baseline	5.77 ± 1.36	5.79 (4.67-6.6)	5.76 ± 1.43	5.6 (4.81-6.42)	0.317	0.761	0.614	0.461
T2	5.78 ± 1.42	5.77 (4.85-6.77)	5.93 ± 1.69	5.53 (4.89-6.9)
T3	5.54 ± 1.54	5.48 (4.19-6.5)	5.76 ± 1.68	5.42 (4.68-6.81)
Cr
Baseline	63.14 ± 14.03	63.55 (53-74.7)	62.33 ± 15.36	61.5 (51.7-69.8)	< 0.0001	0.445	0.202	0.706
T2	67.03 ± 16.09	66.95 (56-77.7)	64.41 ± 15.62	65.85 (54.1-72.4)
T3	67.56 ± 15.11	66 (56.9-75.6)	66.26 ± 15.16	65.6 (56.1-72.7)
SG
Baseline	1.02 ± 0.01	1.02 (1.02-1.03)	1.02 ± 0.01	1.02 (1.02-1.03)	0.424	0.087	0.111	0.030
T2	1.02 ± 0.01	1.02 (1.02-1.03)	1.02 ± 0.01	1.02 (1.01-1.03)
T3	1.03 ± 0.01	1.03 (1.02-1.03)	1.02 ± 0.01	1.02 (1.01-1.03)
Uric acid
Baseline	349.98 ± 82.4	347.75 (303.4-417.9)	346.88 ± 71.75	347.3 (305.6-385.5)	0.582	0.937	0.718	0.887
T2	348.14 ± 71.62	344.7 (294.8-397.5)	340.76 ± 66.08	344.7 (302-377.1)
T3	354.41 ± 78.16	357.2 (302.9-414.5)	349.36 ± 77.77	365.35 (280.2-406.5)
Na
Baseline	140.49 ± 2.1	140.45 (138.9-142.3)	141.01 ± 1.94	140.8 (139.6-142.1)	< 0.0001	0.007	0.005	0.840
T2	140.13 ± 2.15	140.3 (138.9-141.9)	139.34 ± 1.74	139.55 (138.3-140.4)
T3	139.28 ± 2.63	139.55 (137.6-141)	139.7 ± 1.78	139.8 (138.2-140.7)
K
Baseline	4.34 ± 0.32	4.29 (4.14-4.49)	4.25 ± 0.28	4.3 (4.04-4.4)	0.248	0.126	0.851	0.048
T2	4.34 ± 0.33	4.27 (4.12-4.51)	4.26 ± 0.36	4.26 (4.01-4.44)
T3	4.21 ± 0.26	4.2 (4.05-4.31)	4.27 ± 0.36	4.21 (4.06-4.45)
Cl
Baseline	104.64 ± 2.59	104.5 (103-106.2)	104.64 ± 2.34	104.8 (102.9-105.9)	0.002	0.295	0.216	0.988
T2	104.31 ± 2.07	104.55 (102.7-105.5)	103.65 ± 2.06	103.5 (102.6-104.6)
T3	103.55 ± 2.74	104.2 (101.6-105.8)	103.54 ± 2.57	103.55 (101.2-105.6)
ALT
Baseline	28.5 ± 22.64	21.15 (15.9-33.2)	25.32 ± 15.92	19.85 (16.3-28.9)	0.610	0.481	0.473	0.273
T2	28.32 ± 23.09	20.55 (17.3-31.2)	27.67 ± 17.39	22.25 (17.3-33.2)
T3	24.87 ± 11.6	22.25 (14.9-32.9)	26.91 ± 21.95	21 (15.4-29.2)
AST
Baseline	26.3 ± 14.98	21 (17.3-29.9)	23.04 ± 8.84	20.35 (18.2-24.5)	0.609	0.127	0.545	0.057
T2	26.91 ± 18.3	20.5 (18.8-29.1)	25.13 ± 8.54	22.8 (19.6-26.7)
T3	24.4 ± 11.42	20.45 (17.6-28.1)	27.29 ± 17.52	22.15 (18.2-29.4)
ALB
Baseline	45.38 ± 3.28	45.5 (43.7-47.5)	44.41 ± 2.59	44.4 (43-45.7)	0.001	0.364	0.199	0.988
T2	44.38 ± 2.6	44.25 (42.7-46.2)	44.09 ± 2.44	43.6 (42.3-45.5)
T3	44.44 ± 3.1	44.95 (42.4-46.6)	43.46 ± 2.42	43.35 (42.3-44.5)
TBIL
Baseline	16.97 ± 6.72	15.6 (12.72-18.62)	15.84 ± 5.25	15.01 (12.25-17.85)	0.197	0.614	0.324	0.806
T2	15.84 ± 7.29	14 (11.3-18.18)	15.66 ± 4.87	14.83 (12.26-18.15)
T3	16.15 ± 8.76	13.73 (11.24-16.52)	15.23 ± 4.91	13.96 (11.73-16.8)
DBIL
Baseline	2.96 ± 1.36	2.62 (2.13-3.3)	2.68 ± 0.86	2.5 (2.1-3.1)	0.114	0.274	0.108	0.599
T2	2.66 ± 1.17	2.5 (1.88-2.9)	2.66 ± 0.92	2.4 (2.1-3.1)
T3	2.77 ± 1.34	2.4 (1.96-3)	2.57 ± 0.81	2.25 (1.96-3.2)
IBIL
Baseline	14.01 ± 5.67	13.17 (10.76-15.55)	13.16 ± 4.53	12.29 (10.11-15.05)	0.269	0.710	0.412	0.866
T2	13.17 ± 6.29	11.77 (9.5-15.46)	13 ± 4.05	12.32 (10.02-15.57)
T3	13.38 ± 7.66	11.3 (9.44-14.09)	12.66 ± 4.24	11.26 (9.72-14.05)

ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; ALB: Albumin; TBIL: Total bilirubin; DBIL: Direct bilirubin; IBIL: Indirect bilirubin; Cr: Creatinine; SD: Standard deviation; Q1: First quartile; Q3: Third Quartile; T0: Baseline; T2: Second time point of measurement (after 6 weeks); T3: Third time point of measurement (after 12 weeks); SG: Specific gravity.

Table 4 Detailed lipid profile and insulin sensitivity measurements over time

Index	Placebo group (n = 38)		Intervention group (n = 38)		Time (P value)1	Time group (P value)2	T2 vs T0 (P value)	T3 vs T0 (P value)
	mean ± SD	Median (Q1-Q3)	mean ± SD	Median (Q1-Q3)
CHOL
Baseline	5.17 ± 1.16	5.06 (4.48-6.04)	5.16 ± 1.2	5.3 (4.26-5.93)	0.016	0.765	0.797	0.656
T2	4.92 ± 1.22	4.75 (4.12-5.67)	4.85 ± 1.07	4.53 (4.2-5.37)
T3	4.79 ± 1.09	4.74 (4.34-5.63)	4.89 ± 0.88	4.89 (4.32-5.37)
TG
Baseline	2.02 ± 1.95	1.51 (1.14-2.09)	1.91 ± 1.31	1.33 (1.12-2.27)	0.910	0.175	0.226	0.061
T2	2.19 ± 2.36	1.63 (1.04-2.17)	1.73 ± 0.96	1.48 (1.11-1.98)
T3	2.31 ± 2.5	1.45 (1.1-2.37)	1.71 ± 0.85	1.58 (0.97-2.06)
HDL-C
Baseline	1.31 ± 0.32	1.30 (1.06-1.45)	1.24 ± 0.19	1.23 (1.08-1.36)	< 0.0001	< 0.0001	< 0.0001	< 0.0001
T2	1.27 ± 0.30	1.22 (1.07-1.47)	1.36 ± 0.22	1.33 (1.21-1.55)
T3	1.28 ± 0.28	1.27 (1.11-1.47)	1.47 ± 0.24	1.46 (1.26-1.55)
LDL-C
Baseline	3.3 ± 0.85	3.2 (2.86-3.98)	3.35 ± 1	3.62 (2.66-3.89)	0.002	0.806	0.816	0.517
T2	3.07 ± 0.88	3.06 (2.59-3.72)	3.17 ± 0.88	2.84 (2.62-3.69)
T3	2.92 ± 0.85	2.84 (2.55-3.51)	3.09 ± 0.72	3.13 (2.63-3.47)
INS
Baseline	30.08 ± 51.6	11.7 (8-23.72)	16.23 ± 18.4	10.98 (8.6-14.66)	0.051	0.194	0.181	0.181
T2	19.56 ± 23.26	10.4 (8.3-18.72)	14.4 ± 15.39	10.12 (7.8-14.33)
T3	19.59 ± 24.43	11.5 (8.3-18.16)	14.12 ± 14.77	11 (8.7-13.75)
CRP
Baseline	3.86 ± 2.24	3.06 (2.23-4.82)	3.79 ± 2.1 0	3.03 (2.35-3.97)	0.087	0.119	0.290	0.054
T2	3.90 ± 2.70	3.19 (2.22-5.06)	3.37 ± 1.88	2.94 (2.03-4.13)
T3	3.80 ± 2.65	3.07 (1.95-4.42)	2.78 ± 3.02	1.75 (1.17-3.19)
HOMA-IR
Baseline	12.57 ± 21.45	4.53 (2.75-10.24)	6.25 ± 7.38	3.93 (3.01-6.51)	0.376	0.440	0.238	0.503
T2	9.41 ± 14.74	4.51 (2.73-6.29)	6 ± 6.26	3.79 (3.09-5.88)
T3	9.82 ± 19.03	5.15 (2.87-7.8)	5.39 ± 5.15	3.88 (2.77-5.79)
FBG
Baseline	9.20 ± 2.26	8.81 (7.50-10.00)	9.06 ± 1.38	9.06 (7.78-10.18)	< 0.0001	< 0.0001	0.036	0.007
T2	9.17 ± 2.52	8.71 (7.03-9.97)	8.04 ± 1.91	7.83 (6.7-9.03)
T3	9.20 ± 2.56	8.47 (7.41-10.29)	7.34 ± 1.8	7.09 (6.04-8.29)
HbA1c
Baseline	7.95 ± 1.48	7.40 (6.80-9.00)	7.78 ± 1.44	7.60 (6.40-8.50)	0.031	0.901	0.980	0.870
T2	7.85 ± 1.47	7.65 (6.70-8.80)	7.69 ± 1.09	7.75 (6.60-8.30)
T3	7.64 ± 1.46	7.35 (6.70-8.30)	7.44 ± 1.02	7.25 (6.80-8.10)

¹Time (P value): Assesses changes over time within each group.

²Time group (P value): Evaluates the interaction effect between time and group.

CHOL: Cholesterol; TG: Triglycerides; HDL-C: High-density lipoprotein cholesterol; LDL-C: Low-density lipoprotein cholesterol; INS: Insulin; CRP: C-reactive protein; HOMA-IR: Homeostasis model assessment of insulin resistance; FBG: Fasting blood glucose; HbA1c: Glycated hemoglobin; SD: Standard deviation; Q1: First quartile; Q3: Third quartile; T0: Baseline; T2: Second time point of measurement (after 6 weeks); T3: Third time point of measurement (after 12 weeks).

Table 5 Glycemic indicators of participants before and after treatment adjusted for confounders

Variable	Probiotic group (n = 38)	Placebo group (n = 38)	MD (95%CI), P value
FBG (mmol/L)
Baseline	9.06 ± 1.38	9.20 ± 2.26	-0.14 (-0.99-0.70), 0.7422
End	7.34 ± 1.8	9.20 ± 2.56	-1.85 (-2.80 to -0.80), 0.0013
MD (95%CI), P value1	-1.79 (-2.25 to -1.18), 0.001	-0.01 (-0.82 to -0.82), 0.991
HbA1c (%)
Baseline	7.78 ± 1.44	7.90 ± 1.48	-0.16 (-0.80-0.60), 0.6312
End	7.44 ± 1.02	7.64 ± 1.46	-0.20 (-0.78-0.36), 0.9453
MD (95%CI), P value1	-0.34 (-0.70-0.03), 0.071	-0.30 (-0.73-0.13), 0.167
Insulin (mU/mL)
Baseline	16.23 ± 18.40	30.08 ± 51.16	-13.10 (-31.10-3.80), 0.1232
End	14.12 ± 14.77	19.59 ± 24.43	-5.40 (-14.60-3.70), 0.0513
MD (95%CI), P value1	-2.11 (-4.64-0.31), 0.095	-10.49 (-22.80-1.80), 0.093
HOMA-IR index
Baseline	6.25 ± 7.38	12.57 ± 21.46	-6.32 (-13.60-1.10), 0.0902
End	5.39 ± 5.15	9.82 ± 2.10	-4.42 (3.10 to -13.70), 0.0553
MD (95%CI), P value3	-0.85 (-2.10-0.42), 0.182	-2.75 (-8.30-2.08), 0.322

¹P based on paired samples t-test.

²P based on independent samples t-test.

³P based on analysis of covariance adjusted for baseline values, age, duration of diabetes, weight, and calorie intake.

FBG: Fasting blood glucose; MD: Mean difference; HOMA-IR: Homeostasis model assessment indices; HbA1c: Hemoglobin A1c; CI: Confidence interval.