Protective Effect And Mechanism Of Glycosides Of Cistanche in A Rat Cerebral Ischemia Reperfusion Injury Model Ⅱ

2 results

2.1 Effects of GCs on neurological function in CIRI model rats

The Sham group had no neurological deficit symptoms, so it was assigned a score of 0. Compared with the Sham group, the neurological function scores of the Model group, GCs group, and Nim group were significantly increased (P<0.05). Compared with the Model group, neurological function scores were reduced after treatment with GCs and Nim (P<0.05). There was no statistical significance in the comparison between the GCs group and the Nim group (Fig. 1).

Note, Compared with Sham group, * P<0.05. Compared with Model group, # P<0.05. Figure 1 Neurological deficit score

2.2 Effects of GCs on behavior of CIRI model rats

The results of the sticker removal experiment and the balance bar experiment showed that compared with the Sham group, the time for rats to remove the sticker and the time to pass the balance bar were significantly longer after CIRI (P<0.05). Compared with the Model group, the time it took rats to tear off the sticker after treatment with GCs and Nim was significantly reduced (P<0.05), and the time it took to pass the balance bar was shortened, but there was no statistical significance in the comparison between the GCs group and the Nim group (Figure 2A , 2B).
The results of the open field experiment showed that after the model was established, the rats' movement ability was significantly weakened compared with before, the movement trajectories were reduced, the total distance moved was shortened, and the percentage of stationary time increased (P<0.05). Compared with the Model group, the movement ability of rats after treatment with GCs and Nim was improved, the movement trajectory and total distance moved increased, and the immobile time was reduced (P<0.05). There was no statistical significance in the comparison between the GCs group and the Nim group (Fig. 2C~2E).

New Herbal Cistanche Supplements For Cerebral Ischemia-Reperfusion Injury

2.3 Effect of GCs on cerebral infarct size in CIRI model rats

TTC staining detects the area of cerebral infarction. Brain tissue after infarction appears pale and non-infarct appears bright red. The brain tissue of the rats in the Sham group showed a uniform red color after TTC staining, and no infarcts were formed. White infarcts were visible in the Model group and the drug administration group. Compared with the Sham group, the infarct area of rats after cerebral ischemia-reperfusion was significantly increased ( P < 0.05). Compared with the Model group, GCs and Nim could significantly reduce the area of cerebral infarction in model rats (P < 0.05), but there was no significant difference between the two (Figure 3).

Note, A, Time to remove stickers. B, Time passing the balance beam. C, Traveled distance. D, Percentage of immobility time. Compared with Sham group, * P<0.05. Compared with Model group, # P<0.05. Figure 2 Effects of GCs on sensory and motor abilities in rats after CIRI

Note,A, TTC staining. B, Percentage of infarct area. Compared with Sham group, * P<0.05. Compared with Model group, # P<0.05. Figure 3 TTC staining detects the brain infarct area

Cell morphology changes after cerebral ischemia-reperfusion injury. Normal rat cortical nerve cells are neatly arranged, with normal morphological structure, a large number of cells, and uniform and clear Nissl body coloring. When nerve cells undergo necrosis and apoptosis after ischemia and hypoxia, the cell morphology, structure, and number change significantly. The arrangement of nerve cells is disordered, the cell nuclei are shrunk and fragmented, apoptotic bodies can be formed in some parts, the number of Nissl bodies is reduced, and the coloring is light. In this study, both Nissl staining and TUNEL staining results showed that from the perspective of cell morphology, GCs and Nim can reduce the degree of apoptosis and exert a protective effect on nerve cells.

Apoptosis is a process of programmed cell death, which mainly includes endogenous and exogenous pathways. The extrinsic pathway is also called the death receptor pathway. The combination of death ligands (such as TNF-&alpha;, FasL) and death receptors (such as TNFR1, Fas) activates Caspase-8 and then activates Caspase-3 to mediate cell apoptosis [22 ]. The intrinsic pathway is also called the mitochondrial pathway. Mitochondria not only provide energy for cell metabolism, but also play an important role in regulating cell apoptosis [23]. After activation, the Bcl-2 family pro-apoptotic proteins Bax and Bak undergo conformational changes, accumulate in the outer mitochondrial membrane, and directly mediate the permeability of the outer mitochondrial membrane, allowing small molecule substances such as Cyt-c and apoptosis-inducing factors to pass through and Activates the Caspase-3 family, leading to cell apoptosis [24-30]. Bcl-2 can stabilize mitochondrial membrane potential and inhibit the release of Cyt-c to regulate cell apoptosis [31-33]. Zhang Ran [34] found that the opioid receptor agonist DADLE inhibits the activation of Caspase-9 and reduces the expression of Bax/Bcl-2, thereby reducing the apoptosis of brain microvascular endothelial cells and improving their viability. Studies have shown that Cistanche-A can significantly reduce the expression of Caspase and RIPK in the renal tissue of renal ischemia-reperfusion injury model mice, and its protective effect on renal ischemia-reperfusion injury can be

This can be achieved by inhibiting apoptosis [35]. The results of this study show that after treatment with GCs and Nim, the expression of pro-apoptotic proteins Bax and Caspase-3 decreased, and the expression of anti-apoptotic protein Bcl-2/Bax increased, indicating that GCs and Nim can regulate the expression of apoptotic proteins to exert a protective effect. The results of this study show that there is no significant difference in the treatment effects of GCs and Nim on CIRI, indicating that GCs can exert the same effect as Nim.
In summary, GCs can improve the symptoms of neurological deficits in CIRI rats, reduce neurological function scores, shorten sticker removal time and balance bar passing time, improve sensory and motor functions, reduce cerebral infarction area, and downregulate Bax and Caspase-3. expression and upregulate the expression of Bcl-2/Bax. GCs may exert neuroprotective effects by inhibiting apoptosis, thereby improving CIRI.