Sleeve gastrectomy ameliorates endothelial function and prevents lung cancer by normalizing endothelin-1 axis in obese and diabetic rats

Table 1 Specific sequences of primers used in polymerase chain reaction analysis

Gene	Primer sequence, 5’-3’	GenBank No.	Length, bp	Annealing temperature, °C
GAPDH	F: CTGGAGAAACCTGCCAAGTATG	NM_017008.4	138	60
	R: GGTGGAAGAATGGGAGTTGCT
ET-1	F: TCTGCCACCTGGACATCATCTG	NM_012548.2	205	60
	R: CTTTGGGCTCGGAGTTCTTTGT
ET-A	F: ACCGCCATTGAAATTGTCTCC	NM_012550.2	173	60
	R: AGCCACCAGTCCTTCACGTCTT
ET-B	F: CCAAAGACTGGTGGCTGTTCA	XM_006252431.3	183	60
	R: CAAACACGAGGACCAGGCAG
ECE-1	F: CACAACCAAGCCATCATTAAGC	NM_053596.2	275	60
	R: TTGGAGTCGGCACTGACATAGA

F: Forward; R: Reverse.

Table 2 Comparisons of body weight (g) among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
0	401.7 ± 13.0	507.5 ± 20.4	516.6 ± 20.1	< 0.0001	< 0.0001	0.4768
1	421.5 ± 13.9	464.1 ± 21.0	433.0 ± 25.6	< 0.0001	0.3071	0.0003
2	448.8 ± 13.0	480.8 ± 21.6	419.5 ± 27.6	0.0002	0.0006	< 0.0001
4	481.7 ± 13.9	515.3 ± 23.1	437.5 ± 27.0	< 0.0001	< 0.0001	< 0.0001
6	513.8 ± 14.8	555.5 ± 19.9	460.1 ± 30.2	< 0.0001	< 0.0001	< 0.0001
8	540.2 ± 16.1	580.9 ± 19.1	484.1 ± 30.2	< 0.0001	< 0.0001	< 0.0001
10	560.4 ± 10.6	595.2 ± 15.4	510.6 ± 36.5	0.0284	0.0008	< 0.0001
12	578.6 ± 12.1	602.0 ± 11.2	541.4 ± 35.4	0.1953	0.0174	< 0.0001

Data are expressed as the mean ± SD. C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 3 Comparisons of food intake (g/d) among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
0	17.9 ± 2.4	23.9 ± 2.2	24.1 ± 2.2	< 0.0001	< 0.0001	0.9932
1	18.1 ± 2.2	16.8 ± 3.2	11.4 ± 2.7	0.5071	< 0.0001	< 0.0001
2	17.8 ± 2.4	18.0 ± 3.1	11.1 ± 3.4	0.9848	< 0.0001	< 0.0001
4	19.1 ± 2.4	19.8 ± 4.1	12.9 ± 3.8	0.8433	< 0.0001	< 0.0001
6	19.4 ± 3.4	23.2 ± 4.5	13.8 ± 3.6	0.0274	0.0005	< 0.0001
8	20.4 ± 4.5	26.6 ± 3.5	15.2 ± 4.0	< 0.0001	0.0014	< 0.0001
10	21.2 ± 6.2	28.4 ± 1.3	19.0 ± 3.8	0.0018	0.5398	< 0.0001
12	23.0 ± 4.8	32.4 ± 2.7	22.4 ± 3.4	< 0.0001	0.9550	< 0.0001

C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 4 Comparisons of fasting blood glucose (mmol/L) among groups

Time after surgery (wk)					P value
		C	SH	SG	C vs SH	C vs SG	SH vs SG
0	5.3 ± 0.7		15.4 ± 1.5	15.7 ± 1.3	< 0.0001	< 0.0001	0.8776
1	5.5 ± 0.7		12.2 ± 1.6	5.7 ± 1.1	< 0.0001	0.9423	< 0.0001
2	5.7 ± 1.0		12.7 ± 2.3	5.5 ± 1.2	< 0.0001	0.8982	< 0.0001
4	6.2 ± 1.0		13.8 ± 2.7	6.3 ± 1.4	< 0.0001	0.9534	< 0.0001
6	6.3 ± 0.6		14.4 ± 2.7	7.4 ± 2.0	< 0.0001	0.2619	< 0.0001
8	5.9 ± 0.9		16.1 ± 2.4	8.4 ± 1.9	< 0.0001	0.0017	< 0.0001
10	6.2 ± 0.7		20.1 ± 1.6	9.6 ± 1.3	< 0.0001	0.0023	< 0.0001
12	6.0 ± 0.8		22.3 ± 2.7	12.6 ± 1.5	< 0.0001	< 0.0001	< 0.0001

C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 5 Comparisons of areas under curve of oral glucose tolerance test among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
0	956 ± 123	2419 ± 197	2407 ± 147	< 0.0001	< 0.0001	0.9907
4	1148 ± 214	2101 ± 293	1125 ± 226	< 0.0001	0.9651	< 0.0001
8	1241 ± 316	2346 ± 408	1507 ± 314	< 0.0001	0.0591	< 0.0001
12	1353 ± 282	2967 ± 421	2075 ± 247	< 0.0001	< 0.0001	< 0.0001

C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 6 Comparisons of fasting serum insulin levels (mIU/L) among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
0	7.385 ± 1.359	8.243 ± 2.349	6.104 ± 1.734	0.6134	0.2858	0.8341
4	7.989 ± 1.542	8.238 ± 1.597	9.212 ± 2.835	0.0263	0.3189	0.4710
8	10.808 ± 2.309	10.348 ± 1.898	9.946 ± 3.054	0.8770	0.6826	0.9362
12	11.146 ± 2.173	7.734 ± 1.525	9.042 ± 2.005	0.1864	0.3240	0.9439

C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 7 Comparisons of insulin resistance (mIU × mmol/L²) among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
0	1.721 ± 0.157	4.474 ± 1.078	4.216 ± 1.061	< 0.0001	< 0.0001	0.6563
4	2.136 ± 0.234	5.988 ± 1.227	2.503 ± 0.667	< 0.0001	0.4289	< 0.0001
8	2.744 ± 0.309	7.116 ± 0.934	3.525 ± 0.656	< 0.0001	0.0811	< 0.0001
12	2.944 ± 0.365	8.341 ± 0.941	4.979 ± 0.833	< 0.0001	0.0003	< 0.0001

C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 8 Comparisons of endothelin-1 mRNA expression among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
4	1.51 ± 1.21	2.33 ± 1.08	1.31 ± 0.39	0.3293	0.9316	0.1828
8	1.11 ± 0.41	2.88 ± 0.80	0.87 ± 0.39	0.0100	0.9085	0.0033
12	1.56 ± 0.86	3.07 ± 1.55	1.65 ± 0.63	0.0318	0.9878	0.0449

C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 9 Comparisons of endothelin receptor A mRNA expression among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
4	1.17 ± 0.77	1.77 ± 0.88	1.09 ± 0.59	0.4552	0.9859	0.3669
8	1.57 ± 0.42	1.73 ± 0.55	1.21 ± 0.62	0.9460	0.7454	0.5501
12	1.61 ± 0.97	2.92 ± 1.03	2.71 ± 1.01	0.0334	0.0859	0.9047

C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 10 Comparisons of endothelin receptor B mRNA expression among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
4	1.06 ± 0.40	0.96 ± 0.27	0.94 ± 0.58	0.9581	0.9393	0.9981
8	0.98 ± 0.65	1.45 ± 0.62	1.07 ± 0.68	0.3994	0.9666	0.5442
12	1.00 ± 0.53	2.08 ± 0.84	1.17 ± 0.37	0.0141	0.8937	0.0419

C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 11 Comparisons of endothelin-converting enzyme-1 mRNA expression among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
4	1.11 ± 0.61	2.24 ± 0.55	0.99 ± 0.39	0.0191	0.9534	0.0090
8	1.40 ± 0.21	2.35 ± 1.00	1.52 ± 0.57	0.0550	0.9526	0.1031
12	1.32 ± 0.81	2.60 ± 0.73	1.65 ± 0.35	0.0073	0.6881	0.0552

C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 12 Comparisons of relative endothelin-1 protein expression among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
4	0.95 ± 0.11	1.10 ± 0.14	0.69 ± 0.10	0.3229	0.0354	0.0007
8	0.68 ± 0.10	1.79 ± 0.25	1.02 ± 0.12	< 0.0001	0.0056	< 0.0001
12	0.60 ± 0.06	1.64 ± 0.20	1.36 ± 0.24	< 0.0001	< 0.0001	0.0229

C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 13 Comparisons of relative endothelin receptor A protein expression among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
4	0.87 ± 0.17	0.82 ± 0.13	0.61 ± 0.08	0.7836	0.0036	0.0207
8	0.72 ± 0.16	0.97 ± 0.13	0.82 ± 0.08	0.006	0.44	0.1108
12	0.50 ± 0.09	1.17 ± 0.11	1.03 ± 0.09	< 0.0001	< 0.0001	0.1867

C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 14 Comparisons of relative endothelin receptor B protein expression among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
4	1.06 ± 0.11	1.29 ± 0.16	1.26 ± 0.20	0.0575	0.095	0.9697
8	0.54 ± 0.11	1.71 ± 0.16	1.03 ± 0.14	< 0.0001	< 0.0001	< 0.0001
12	1.14 ± 0.08	2.28 ± 0.19	1.00 ± 0.16	< 0.0001	0.2966	< 0.0001

C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 15 Comparisons of relative endothelin-converting enzyme-1 protein expression among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
4	0.13 ± 0.03	0.36 ± 0.06	0.39 ± 0.02	< 0.0001	< 0.0001	0.5966
8	0.15 ± 0.04	0.25 ± 0.05	0.24 ± 0.04	0.0123	0.0125	> 0.9999
12	0.19 ± 0.05	0.37 ± 0.06	0.22 ± 0.06	< 0.0001	0.6298	< 0.0001

C: Control; SH: Sham; SG: Sleeve gastrectomy.

Table 16 Comparisons of semi-quantified DNA damage levels (percentage of γ-H2AX-positive cells) among groups

Time after surgery (wk)				P value
	C	SH	SG	C vs SH	C vs SG	SH vs SG
4	0.83 ± 0.43	3.06 ± 0.47	1.15 ± 0.31	0.0009	0.835	0.0046
8	1.71 ± 0.63	4.73 ± 1.04	1.73 ± 0.72	< 0.0001	0.999	< 0.0001
12	1.86 ± 0.37	6.70 ± 1.80	2.75 ± 1.09	< 0.0001	0.2672	< 0.0001

C: Control; SH: Sham; SG: Sleeve gastrectomy.