Glucagon-like peptide (GLP) -2 improved colonizing bacteria and reduced severity of ulcerative colitis by enhancing the diversity and abundance of intestinal mucosa

Figures & data

Table 1. Patients demographics and clinical characteristics

Number of (patients No.)	Gender	Age	Height	Weight	Medical history	Relative normal	Critical point	Diseased (Dissected) region*	Terminal ileum**	Extent of disease	Clinical type
3	female	47	162	55 kg	NO	3 C	3D	3E	3A	0–30	Mild
4	male	51	176	64 kg	NO	4B	4D	4E	4A	0–60	Severe
5	male	52	174	58 kg	NO	5 C	5B	5A	5D	0–45	Severe
12	female	65	162	54 kg	NO	12B	12 C	12D	12A	0–50	Mild
13	male	49	176	70 kg	NO	13B	13 C	13E	13A	0–45	Mild
14	male	34	178	68 kg	NO	14B	14 C	14E	14A	0–25	Severe
16	female	33	163	58 kg	NO	16B	16 C	16D	16A	0–70	Mild
Normal (control)								Rectum	Terminal ileum
40	female	61	162	55 kg	NO	NA	NA	40B	40A	NA	NA
41	female	44	165	68 kg	NO	NA	NA	41B	41A	NA	NA
42	female	49	174	70 kg	NO	NA	NA	42B	42A	NA	NA
43	male	55	178	68 kg	NO	NA	NA	43B	43A	NA	NA
44	male	69	175	74 kg	NO	NA	NA	44B	44A	NA	NA

*Atlas of … .

**Atlas of … .

Figure 1. α Diversity analysis A, B, C for Sobs, Shannon, and ACE indices of UC patients and healthy control subjects at the OTU level. D, E, F for Significant differences between samples from 2 selected groups. *P < 0.05, **P < 0.01, ***P < 0.001. X and Y axes show group names and exponential averages for each group, respectively

Figure 2. Microbiota composition,A, B, C represent the Venn diagram with different colors representing different groups (or samples); numbers in overlapping areas represent the number of species common to multiple groups, and numbers in nonoverlapping areas represent the number of unique species in corresponding groups. D, the X and Y axes show sample or group names and species names, respectively. The abundances of different species in the samples are represented by a color gradient; values represented by the color are shown to the right. E, F represent the Circos diagram of the relationship between sample and species

Figure 3. β diversity analysis, A, different colors represent sample groupings, and the box chart in the figure represents the dispersion of different groups of samples on the PC1 axis. B, X and Y axes show 2 selected spindles, and the percentages are the interpreted value of the axes for differences in sample composition. The scales of the X and Y axes are relative distances that have no practical significance. Different colors or shapes represent different groupings of samples; a shorter distance between 2 sample points indicates greater similarity in their species composition. C, different color nodes represent microbiota that is significantly enriched in the corresponding group and have a significant effect on intergroup differences; pale yellow nodes represent microbiota that are not significantly different across groups or that have no significant effect on intergroup differences

Figure 3. Continued

Figure 4. Relationship between species composition and lesion severity. Compared to normal control samples, UC patients showed reduced microbiota diversity and abundance in the colon irrespective of lesion severity, with significant differences observed at the phylum level (Actinobacteriota, Chloroflex, and Patescibacteria) and genus level (Ralstonia and Rhodococcus, which were less abundant in UC patients) (a, b). Lowerdiversity and abundance of microbiota was also observed in the small intestine of UC patients, with significant differences for Acidobacteria, Planctomycetota, and other phyla (c, d). Among UC samples, there were differences between relatively normal tissue and lesions at the phylum (e.g., Actinobacteriota, and Chloroflexi) and genus (eg, Ralstonia, Rhodococcus, Burkholderia-Caballeronia-Paraburkholderia) levels (e, f). Thus, differences in intestinal mucosa microbiome profiles at the phylum and genus levels in UC patients were independent of lesion severity (g, h)

Table 2. Optical density values for serum GLP-2 detected by ELISA in ulcerative colitis patients and healthy control subjects

Group^†	N	Mean^‡	Standard deviation	Standard error of the mean
1	11	0.32223636	0.100551340	0.030317370
2	9	0.51012778	0.112026674	0.037342225

^†Group 1, ulcerative colitis patients; Group 2, healthy control subjects.

^‡Values for both groups conformed to a normal distribution, and statistical significance was evaluated with the independent samples t-test(p < 0.05) between groups 1 and 2.

ELISA, enzyme-linked immunosorbent assay; GLP-2, glucagon-like peptide 2.

Table 3. Results of the independent samples test for optical density value of serum GLP-2 in ulcerative colitis patients and healthy control subjects

	Levene’s test of variance		t-test of mean
	F	Significance	t	df	P-value (2-sided)	Mean difference	Standard error	95% CI of mean difference
Equal variance assumed	0.501	0.488	−30.951	18	0.001	−0.187891414	0.047555924	−0.087980124, −0.287802704
Unequal variance assumed			−30.906	160.342	0.001	−0.187891414	0.048099737	−0.086097686, −0.289685143

CI, confidence interval; GLP-2, glucagon-like peptide 2.

Table 4. The expression of Glp-2 in colonic mucosa group number (-) (+) (++) (+++) UC 11 4 3 3 1 Normal 9 0 2 4 3

Chi-squared Test
	value	df	ProgressiveSig. (bilateral)	Precision Sig.(bilateral)	Precise Sig.(unilateral)
Pearson chi-square	16.000a	1	.000
continuous	12.195	1	.000
Correction Likelihood ratio	21.930	1	.000
Fisher’s exact test				.000	.000
Linear and linear combination	15.000	1	.000
N in a valid case	16

Note: Because the expected count was less than 5, the sample size was insufficient, but Fisher’s exact probability method could be used to test, and the positive rate difference between the two groups was statistically significant (P = 0.00).

Figure 5. Immunohistochemical detection of GLP-2 expression in the intestinal mucosa. A, strong positive expression of GLP-2 (+++) (most healthy control samples). B, weak positive expression of GLP-2 (+). C, negative expression of LP-2 (−) (most UC samples), D, diagnosed UC sample histology H&E staining

Figure 6. [α diversity]. a X and Y axes show clinical parameters and bacterial species present in the intestinal mucosa, respectively. *P < 0.05, **P < 0.01, ***P < 0.001. b–f X and Y axes show clinical parameters and β – or α-diversity index, respectively. The X-coordinates represent environmental factors (e.g., pH, temperature, etc.), the Y-coordinates represent the rank of β-diversity or the α-diversity index, R2 is a determining factor, representing the proportion of variation interpreted by the linear regression line, and the greater R2 index, the higher the degree of interpretation of this environmental factor is on the differences in the composition of the group or in the α-diversity index. b C-reactive protein. c Erythrocyte sedimentation rate. d Thrombocyte count. e GLP-2. f OGTT