[Protection of penehyclidine hydrochloride on renal tissue injury induced by limb ischemia/reperfusion].

OBJECTIVE: To evaluate the protection of penehyclidine hydrochloric postconditioning on HIF-1α (hypoxia-inducible factor-1α) in renal tissue injury induced by lower limb ischemia/reperfusion (I/R). METHODS: A total of 72 adult male Wistar rats weighing 230 - 250 g were randomly divided into 3 groups: control (group C), limb ischemia-reperfusion (group R/I) and penehyclidine hydrochloride postconditioning (group P). The animals were anesthetized by inhaling 2% isoflurane and blood flow of bilateral lower limbs was blocked with rubber bands for 3 h in groups P and R/I. In group P, penehyclidine hydrochloride 0.15 mg/kg was injected via caudal vein at 3 min pre-reperfusion. After sacrificing, their kidneys were removed at 3 h of ischemia and 1, 3, 6 h of reperfusion respectively. The blood urea nitrogen (BUN) and creatinine (Cr) were detected by colorimetric method, plasma tumor necrosis factor-α (TNF-α) by ELISA (enzyme-linked immunosorbent assay) and HIF-1α of renal tissue by immunohistochemistry. Renal pathological changes were observed under light microscope. RESULTS: Compared with group C, the serum levels of BUN and Cr increased while TNF-α and HIF-1α were up-regulated in groups I/R and P (P < 0.05). As compared with group I/R, the serum levels of BUN, Cr and MDA decreased while TNF-α and HIF-1α were down-regulated in group P. [at T2: (15.10 ± 1.88) mmol/L vs (19.46 ± 2.76) mmol/L, (113 ± 10) µmol/L vs (143 ± 11) µmol/L, (13.8 ± 1.7) nmol/g vs (15.5 ± 1.8) nmol/g, (53.1 ± 3.1) ng/L vs (53.9 ± 4.8) ng/L, 0.298 ± 0.015 vs 0.471 ± 0.032, all P < 0.05]. CONCLUSION: Penehyclidine hydrochloride can down-regulate the expression of HIF-1α and attenuate the renal injury induced by lower limb I/R. And the mechanisms may be through inhibiting the inflammatory reactions, reducing the release of oxygen free radicals and improving the conditions of hypoxia and ischemia.

[Preventive effects of ischemic postconditioning and penehyclidine hydrochloride on gastric against ischemia-reperfusion injury in rats].

OBJECTIVE: To investigate the protective effects of ischemic postconditioning and penehyclidine hydrochloride on gastric injury induced by ischemia-reperfusion of lower limb in rats. METHODS: The model of limb ischemia reperfusion injury was used to perform this experiment. One hundred and forty four male Wistar rats weighing 220 - 250 g were randomly divided into 4 groups: group I Control (C), group II Ischemic Reperfusion (IR), group III Ischemic postconditioning (IPO) and group IV penehyclidine hydrochloride (IPHC); C, IR, IPO and IPHC groups has been followed for 0(T(0)), 1(T(1)), 3(T(3)), 6(T(6)), 12(T(12)), or 24(T(24)) perfusion, all the groups were secondary separated into six subgroups as time point and each subgroup contained six rats, respectively. Blood samples from the inferior vena cava were taken for determination of LDH, CK activities and TNF-α, IL-10 content at every time point of reperfusion;the animals were killed at every time point respectively and the gastric were removed for determination of SOD, MPO, XOD and LDH activities, MDA content, and histological examination and the expression of HIF-1α was analyzed. RESULTS: Compared with group C, IR, IPO and IPHC, in serum LDH and CK activities were increased, TNF-α and IL-10 content were increased (P < 0.05 or P < 0.01); and in gastric tissue MPO, XOD and LDH activities were increased and MDA content increased, while SOD activity decreased in group IR, IPO and IPHC (P < 0.05 or P < 0.01); and gastric tissue resulted in significant injury as evidenced by infiltrated of few neutrophils or eosinophils and rare neutrophils between the gastric mucosa or muscularis mucosa and the glands, interstitial vascular dilation hyperemia and small quantity hemorrhage from deep layers of mucosa or interstitial vascular dilation hyperemia, and the expression of HIF-1α was significantly increased (P < 0.01). Compared with group IR, IPO and IPHC in serum LDH and CK activities, TNF-α content decreased while IL-10 content were increased (P < 0.01); and in gastric tissue MDA content, MPO, XOD and LDH activities were decreased, and SOD activity increased in group IPO and IPHC (P < 0.05 or P < 0.01); and the histological injury were milder and the expression of HIF-1α was significantly decreased (P < 0.01). Compared with group IPO, IPHC, in serum LDH activities were makable decreased, CK activities were first increased and then declined, TNF-α content makable declined while IL-10 content were increased (P < 0.05 or P < 0.01), and the histological injury were milder and the expression of HIF-1α was makable decreased (P < 0.01) and in gastric tissue SOD activity were makable increased, MPO activities significantly decreased, MDA content increased at T(3), XOD activities increased after T(12), LDH activities increased at T(3) and declined after T(12) in group IPHC (P < 0.05 or P < 0.01). CONCLUSION: The combination of ischemia postconditioning and postconditioning with penehyclidine hydrochloride can protect the gastric from ischemia-reperfusion injury induced by ischemia reperfusion of the lower limbs in rats, the main mechanism may be reducing post-ischemic oxidative damage, inflammatory reaction, amelio-rating microcirculatory and cellular energy metabolism et al. Additionally, this study found that the protective effects of penehyclidine hydrochloride on gastric injury induced by ischemia reperfusion of the lower limbs, were better than ischemic postconditioning, and the mechanism might be related to its anti-inflammatory effect, antioxidant action and prevention of cell injury et al.

Oxidative stress and hypoxia-induced factor 1α expression in gastric ischemia.

AIM: To investigate the relation of reactive oxygen species (ROS) to hypoxia induced factor 1α (HIF-1α) in gastric ischemia. METHODS: The animal model of gastric ischemia reperfusion was established by placing an elastic rubber band on the proximal part of the bilateral lower limb for ligature for 3 h and reperfusion for 0, 1, 3, 6, 12 or 24 h. Ischemic post-conditioning, three cycles of 30-s reperfusion and 30-s femoral aortic reocclusion were conducted before reperfusion. Histological and immunohistochemical methods were used to assess the gastric oxidative damage and the expression of HIF1-α in gastric ischemia. The malondialdehyde (MDA) content and superoxide dismutase (SOD), xanthine oxidase (XOD) and myeloperoxidase (MPO) activities were determined by colorimetric assays. RESULTS: Ischemic post-conditioning can reduce post-ischemic oxidative stress and the expression of HIF-1α of gastric tissue resulting from limb ischemia reperfusion injury. MDA, SOD, XOD and MPO were regarded as indexes for mucosal injuries from ROS, and ROS was found to affect the expression of HIF-1α under gastric ischemic conditions. CONCLUSION: ROS affects HIF-1α expression under gastric ischemic conditions induced by limb ischemia reperfusion injury. Therefore, ROS can regulate HIF-1α expression in gastric ischemia.

Effects of penehyclidine hydrochloride in small intestinal damage caused by limb ischemia-reperfusion.

AIM: To investigate the protective effect of penehyclidine hydrochloride post-conditioning in the damage to the barrier function of the small intestinal mucosa caused by limb ischemia-reperfusion (LIR) injury. METHODS: Male Wistar rats were randomly divided into three groups (36 rats each): the sham-operation group (group S), lower limb ischemia-reperfusion group (group LIR), and penehyclidine hydrochloride post-conditioning group (group PHC). Each group was divided into subgroups (n = 6 in each group) according to ischemic-reperfusion time, i.e. immediately 0 h (T1), 1 h (T2), 3 h (T3), 6 h (T4), 12 h (T5), and 24 h (T6). Bilateral hind-limb ischemia was induced by rubber band application proximal to the level of the greater trochanter for 3 h. In group PHC, 0.15 mg/kg of penehyclidine hydrochloride was injected into the tail vein immediately after 3 h of bilateral hind-limb ischemia. The designated rats were sacrificed at different time-points of reperfusion; diamine oxidase (DAO), superoxide dismutase (SOD) activity, myeloperoxidase (MPO) of small intestinal tissue, plasma endotoxin, DAO, tumor necrosis factor-α (TNF-α), and interleukin (IL)-10 in serum were detected in the rats. RESULTS: The pathological changes in the small intestine were observed under light microscope. The levels of MPO, endotoxin, serum DAO, and IL-10 at T1-T6, and TNF-α level at T1-T4 increased in groups LIR and PHC (P < 0.05) compared with those in group S, but tissue DAO and SOD activity at T1-T6 decreased (P < 0.05). In group PHC, the tissue DAO and SOD activity at T2-T6, and IL-10 at T2-T5 increased to higher levels than those in group LIR (P < 0.05); however, the levels of MPO, endotoxin, and DAO in the blood at T2-T6, and TNF-α at T2 and T4 decreased (P < 0.05). CONCLUSION: Penehyclidine hydrochloride post-conditioning may reduce the permeability of the small intestines after LIR. Its protection mechanisms may be related to inhibiting oxygen free radicals and inflammatory cytokines for organ damage.