Biochemical and histopathological evidence on the beneficial effects of Tragopogon graminifolius in TNBS-induced colitis

Figures & data

Table 1. Criteria for scoring macroscopic and microscopic colonic damage.

Score for macroscopic damage	Score for microscopic damage
0: Normal appearance with no damage	0: No damage
1: Localized hyperemia without ulceration	1: Focal epithelial edema and necrosis
2: Linear ulceration without significant inflammation	2: Disperse swelling and necrosis of the villi
3: Linear ulceration with inflammation at one site	3: Necrosis with neutrophil infiltration in submucosa
4: Two or more sites of ulceration and extending more than 1 cm   along the length of colon	4: Wide spread necrosis with massive neutrophil infiltration and hemorrhage
5–8: Damage extending more than 2 cm along, the length of colon   and the score is enhanced by 1 for each increased cm of   involvement

Figure 1. Histological images of colon tissues obtained from control and experimental groups. Microscopic evaluation of the control group showed intense transmural inflammation and/or diffuse necrosis (A) and severe crypt destruction (B). In histological examination of the dexamethasone group, minimal mucosal inflammation was observed (C). In the TG-20 group, almost complete necrosis of crypts (D) and severe inflammation of mucosa and submucosa were observed (E). In the TG-30 group, diffuse destruction of crypts was observed (F). In the TG-50 group, no necrosis in crypt (G) and mild inflammation of mucosa (H) were observed.

Table 2. Macroscopic and microscopic scores as criteria for assessing colonic damage.

	Macroscopic score		Microscopic score
Group	Mean ± SEM	Median (min–max)	Mean ± SEM	Median (min–max)
Normal	0.00 ± 0.00	0 (0–0)	0.00 ± 0.00	0 (0–0)
Control	6.22 ± 0.84^a	6 (5–7)	3.83 ± 0.41^a	4 (3–4)
Dexa	0.61 ± 0.55^a,b	1 (0–1)	1.57 ± 0.29^a,b	1 (1–3)
TG-20	3.82 ± 0.55^a,b,c	3 (2–4)	2.33 ± 0.52^a,b,c	2 (2–3)
TG-30	1.70 ± 0.49^a,b,c	2 (1–3)	2.14 ± 0.52^a,b,c	2 (1–3)
TG-50	0.43 ± 0.53^a,b	0 (0–1)	1.67 ± 0.52^a,b	2 (1–2)

Values are mean ± SEM. Dexa, dexamethasone; TG-20, T. graminifolius at dose of 20 mg/kg; TG-30, T. graminifolius at dose of 30 mg/kg; TG-50, T. graminifolius at dose of 50 mg/kg.

^aSignificantly different from the normal group at p < 0.05.

^bSignificantly different from the control group at p < 0.05.

^cSignificantly different from the dexamethasone group at p < 0.05.

Figure 2. Tumor necrosis factor-alpha (TNF-α) level in colon. Values are mean ± SEM. Dexa, dexamethasone; TG20, T. graminifolius at dose of 20 mg/kg; TG30, T. graminifolius at dose of 30 mg/kg; TG50, T. graminifolius at dose of 50 mg/kg. ^aSignificantly different from the Normal group at p < 0.05. ^bSignificantly different from the control group at p < 0.05. ^cSignificantly different from the Dexa group at p < 0.05. ^aaSignificantly different from the Normal group at p < 0.01. ^bbSignificantly different from the control group at p < 0.01. ^ccSignificantly different from the Dexa group at p < 0.01. ^aaaSignificantly different from the Normal group at p < 0.001. ^bbbSignificantly different from the control group at p < 0.001. ^cccSignificantly different from the Dexa group at p < 0.001.

Figure 3. Interleukin 1-beta (IL-1β) level in colon. Values are mean ± SEM. Dexa, dexamethasone; TG20, T. graminifolius at dose of 20 mg/kg; TG30, T. graminifolius at dose of 30 mg/kg; TG50, T. graminifolius at dose of 50 mg/kg. ^aSignificantly different from the Normal group at p < 0.05. ^bSignificantly different from the control group at p < 0.05. ^cSignificantly different from the Dexa group at p < 0.05. ^aaSignificantly different from the Normal group at p < 0.01. ^bbSignificantly different from the control group at p < 0.01. ^ccSignificantly different from the Dexa group at p < 0.01. ^aaaSignificantly different from the Normal group at p < 0.001. ^bbbSignificantly different from the control group at p < 0.001. ^cccSignificantly different from the Dexa group at p < 0.001.

Figure 4. Total antioxidant power as ferric-reducing antioxidant power (FRAP) level in colon. Values are mean ± SEM. Dexa, dexamethasone; TG20, T. graminifolius at dose of 20 mg/kg; TG30, T. graminifolius at dose of 30 mg/kg; TG50, T. graminifolius at dose of 50 mg/kg. ^aSignificantly different from the Normal group at p < 0.05. ^bSignificantly different from the control group at p < 0.05. ^cSignificantly different from the Dexa group at p < 0.05. ^aaSignificantly different from the Normal group at p < 0.01. ^bbSignificantly different from the control group at p < 0.01. ^ccSignificantly different from the Dexa group at p < 0.01. ^aaaSignificantly different from the Normal group at p < 0.001. ^bbbSignificantly different from the control group at p < 0.001. ^cccSignificantly different from the Dexa group at p < 0.001.

Figure 5. Lipid peroxidation (LPO) levels as thiobarbituric acid reacting substances (TBARS) in colon. Values are mean ± SEM. Dexa, dexamethasone; TG20, T. graminifolius at dose of 20 mg/kg; TG30, T. graminifolius at dose of 30 mg/kg; TG50, T. graminifolius at dose of 50 mg/kg. ^aSignificantly different from the Normal group at p < 0.05. ^bSignificantly different from the control group at p < 0.05. ^cSignificantly different from the Dexa group at p < 0.05. ^aaSignificantly different from the Normal group at p < 0.01. ^bbSignificantly different from the control group at p < 0.01. ^ccSignificantly different from the Dexa group at p < 0.01. ^aaaSignificantly different from the Normal group at p < 0.001. ^bbbSignificantly different from the control group at p < 0.001. ^cccSignificantly different from the Dexa group at p < 0.001.

Figure 6. Myeloperoxidase activity (MPO) in colon. Values are mean ± SEM. Dexa, dexamethasone; TG20, T. graminifolius at dose of 20 mg/kg; TG30, T. graminifolius at dose of 30 mg/kg; TG50, T. graminifolius at dose of 50 mg/kg. ^aSignificantly different from the Normal group at p < 0.05. ^bSignificantly different from the control group at p < 0.05. ^cSignificantly different from the Dexa group at p < 0.05. ^aa Significantly different from the Normal group at p < 0.01. ^bbSignificantly different from the control group at p < 0.01. ^ccSignificantly different from the Dexa group at p < 0.01. ^aaaSignificantly different from the Normal group at p < 0.001. ^bbbSignificantly different from the control group at p < 0.001. ^cccSignificantly different from the Dexa group at p < 0.001.

Figure 7. Chemical structures of the phenolic compounds that are quantified by HPLC for standardization of the extract from T. graminifolius.