Effect of Lupinus rotundiflorus gamma conglutin treatment on JNK1 gene expression and protein activation in a rat model of type 2 diabetes

Figures & data

Table 1. Primer sequences and PCR conditions used in qRT-PCR.

Gene	Primer sequence		UPL number	Annealing temperature (°C)	Elongation time (s)
Jnk1	Fw	5′-GAGCACACAATAGAGGAGTGGA-3′	25	57	1
	Rev	5′-CCCTCTTATGACGCCATTCT-3′
Rps18	Fw	5′-CAGAAGGACGTGAAGGATGG-3′	1	57	1
	Rev	5′-TCTATGGGCTCGGATTTTCTT-3′

Figure 1. Analysis of L. rotundiflorus seed extracts using 12% SDS-PAGE. Representative SDS-PAGE analysis of 5 experiments. Molecular weight marker (line 1), Cγ of L. rotundiflorus under native (line 2), and denaturing conditions (line 3).

Table 2. Pre- and post-treatment levels of serum biochemical parameters.

Groups	Ctrl	HG-Ctrl	HG-Met	HG-Lr
Glucose (mg/dL)
Pre	69.30 ± 5.90	308.80 ± 44.60	304.00 ± 23.30	331.30 ± 27.60
Post	62.00 ± 3.20	301.60 ± 56.60	229.20 ± 28.30*	245.00 ± 53.30*
Creatinine (mg/dL)
Pre	0.80 ± 0.03	0.76 ± 0.02	0.83 ± 0.02	0.80 ± 0.03
Post	0.90 ± 0.03	0.76 ± 0.02	0.84 ± 0.04	0.77 ± 0.02
Urea (mg/dL)
Pre	62.17 ± 1.80	58.80 ± 6.26	66.56 ± 5.38	79.33 ± 5.24
Post	57.17 ± 2.20	55.60 ± 3.19	67.44 ± 3.95	72.00 ± 4.04
AST (U/L)
Pre	136.33 ± 9.50	144.60 ± 14.07	100.25 ± 9.06	119.83 ± 6.39
Post	155.00 ± 6.60	147.20 ± 18.41	127.60 ± 8.95*	125.50 ± 7.68
ALT (U/L)
Pre	44.50 ± 2.00	67.60 ± 3.34	63.17 ± 4.41	78.83 ± 3.61
Post	51.67 ± 2.80	66.80 ± 5.78	71.67 ± 3.17	81.00 ± 5.75

Data are expressed as mean ± SEM (n = 5 per group). *p ˂ 0.05 comparing post- vs. pre-treatment levels. ALT: alanine aminotransferase; AST: aspartate aminotransferase; Ctrl: control group, nondiabetic rats without treatment; HG-Ctrl: diabetic rats administered with vehicle (saline); HG-Lr: diabetic rats treated with Cγ from L. rotundiflorus; HG-Met: diabetic rats treated with metformin; Pre: serum levels before treatment; Post: serum levels after one week of treatment.

Figure 2. Relative mRNA expression of Jnk1 in the liver of treated and untreated rats with experimental type 2 diabetes. Data are expressed as mean ± SEM of five rats per group. *p ˂ 0.05, **p ˂ 0.01 when compared with HG-Ctrl group; ^δp ˂ 0.05 when compared with Ctrl group. Ctrl, control group, nondiabetic rats without treatment; HG-Ctrl, diabetic rats administered with vehicle (saline); HG-Lr, diabetic rats treated with Cγ from L. rotundiflorus; HG-Met, diabetic rats treated with metformin.

Figure 3. Western blot analysis of phosphorylated JNK1 in the liver of treated and untreated rats with experimental type 2 diabetes. (A) Representative blots are shown for each group. (B) Relative quantification of phosphorylated JNK1 expressed as phosphorylated JNK1/total JNK1 ratio, using β-actin as a loading control. Values represent mean ± SEM of five rats per group. Ctrl, control group, nondiabetic rats without treatment; HG-Ctrl, diabetic rats administered with vehicle (saline); HG-Lr, diabetic rats treated with Cγ from L. rotundiflorus; HG-Met, diabetic rats treated with metformin; pJNK1, phosphorylated JNK1.