Adiponectin, but Not TGF-β1, CTGF, IL-6 or TNF-α, May Be a Potential Anti-Inflammation and Anti-Fibrosis Factor in Keloid

Figures & data

Table 1 Primers Used in qPCR

Gene	Forward (5ʹ-3ʹ)	Reverse (5ʹ-3ʹ)	Product Size (bp)
Adiponectin	GGAGAACCTGGAGAAGGTGC	GTACAGCCCAGGAATGTTGC	17lbp
TGF-ß 1	CACCCAGCCCACGAATCATCTCC	GGGCCCCCTTCTCTCTACCTG	185bp
CTGF	GCGAAGCTGACCTGGAAGAGAAC	GGCCGTCGGTACATACTCCAC	147bp
IL-6	TGTGTGAAAGCAGCAAAG	AGTCTCCTCATTGAATCCA	101bp
TNF-α	TGTAGCCCATGTTGTAGCAAACC	GAGGACCTGGGAGTAGATGAGGTA	158bp
Collagen I	AGCCAGCAGATCGAGAACAT	TCCTTGGGGTTCTTGCTGAT	245bp
FN	GCCAGATGATGAGCTGCAC	GAGCAAATGGCACCGAGATA	142bp
MMP-1	GTGTGGAGTGCCTGATGTG	CTGCTTGACCCTTGGAGAC	293bp
GAPDH	GAAGGTGAAGGTCGGAGTC	GAAGATGGTGATGGGATTTC	226bp

Table 2 Comparison of Clinic Data Between Patients with Keloids and Controls

Clinic Data	Patients with Keloids (n=50)	Controls (n=50)	P value
Gender
Male	23	25	0.231
Female	27	25	0.184
Age	31.88 ± 8.83	32.27 ± 9.41	0.246
BMI	21.32 ± 2.56	22.91 ± 3.06	0.443
SBP	121.68 ± 13.25	127.95 ± 17.74	0.782
DBP	79.58 ± 9.52	82.09 ± 8.42	0.691
FBG	4.36 ± 0.79	5.32 ± 1.44	0.524
TG	1.35 ± 0.43	1.27 ± 0.58	0.585
TC	4.73 ± 1.28	5.04 ± 0.93	0.854
Family history of Keloid	19	2	< 0.001

Abbreviations: BMI, body mass index, kg/m²; SBP, systolic blood pressure, mmHg; DBP, diastolic blood pressure, mmHg; FBG, fasting blood glucose, mmol/L; TG, triglyceride, mmol/L; TC, cholesterol, mmol/L.

Table 3 To Compare the Expression of Adiponectin, TGF-β1, CTGF, IL-6 and TNF-α in Peripheral Blood Between Patients with Keloids and Controls

Cytokines	Patients with Keloids (n=50)	Controls (n=50)	P value
Adiponectin (μg/mL)	2.06 ± 0.99	8.95 ± 2.66	< 0.01
TGF-ß1 (pg/mL)	928.44 ± 200.02	408.10 ± 104.69	< 0.01
CTGF (ng/mL)	8.71 ± 3.02	2.72 ± 1.19	< 0.01
IL-6 (pg/mL)	117.60 ± 36.82	47.46 ± 15.28	< 0.01
TNF-a (pg/mL)	92.27 ± 21.49	39.65 ± 9.91	< 0.01

Figure 1 The serum concentrations of adiponectin (A), TGF-β1 (B), CTGF (C), IL-6 (D) and TNF-α (E) in normal controls and tissues from keloid were detected by ELISA. Data represent three independent experiments. **P<0.01 compared with the control group.

Figure 2 The mRNAs expression of adiponectin (A), TGF-β1 (B), CTGF (C), IL-6 (D) and TNF-α (E) in normal controls and tissues from keloid was detected by qPCR. Data represent three independent experiments.*P<0.05 versus the control group.

Figure 3 Correlation analysis between serum concentration of adiponectin with VSS scores (A) and ages (B) in keloid patients were measured by Pearson’s correlation. And the serum expression levels of adiponectin between different genders in patients with keloid were examined by t test (C).

Figure 4 Adiponectin attenuated TGF-β1 mediated expression of collagen I and FN, and upregulates TGF-β1 suppressed MMP-1 expression in KFs by AMPK signaling pathway. KFs and NFs were preincubated with Compound C (10 µM, an AMPK inhibitor), or LY294002 (10 µM, a PI3K inhibitor) or without inhibitors for 24 h. After stimulation with or without adiponectin (APN, 5 µg/mL) in the presence of TGF-β1 (10 ng/mL) for 24 h, the supernatant concentration and mRNA levels of type I collagen (A and D), FN (B and E), and MMP-1 (C and F) in KFs and NFs were examined by ELISA and qPCR, respectively. Data represent at least three independent experiments. *P<0.05 versus the control group, **P<0.01 versus the control group; ^#P<0.05 versus the TGF-β1 group, ^##P<0.01 versus the TGF-β1 group; ^&P<0.05 versus the TGF-β1+adiponectin group, ^&&P<0.01 versus the TGF-β1+adiponectin group. The results are shown as means±SE.