The Effects Of Heshouwuyin On Proliferation And Apoptosis In Kidney Cortex Of Aging Rats

Recently, the research shows that renal diseases of aging people are closely related with the senescence of kidney and the deterioration of the function and the relationship between proliferation and apoptosis in kidney cortex plays an important role in renal senescence and diseases. Heshouwu was selected according to the theory of chinese traditional medicine. Sub-acutely aging rats were induced by D-galactose. SP immunohistochemistry was used to observe the expression of PCNA, Bax, Bcl-2, Fas, Fasl and Caspase-3 and Tdt-mediated dUTP nick end labelling (TUNEL) was used to observe the expression of apoptosis in kidney cortex. Bax and bcl-2 proteins were detected by Western Bloting. The effects of Heshouwuyin on proliferation and apoptosis was observed in kidney cortex of aging rats and the therapeutic mechanism was explored. The study will provide experimental bases for choosing anti -aging traditional chinese medicine.Objective:To observe the effects of traditional Chinese medicine Heshouwuyin on proliferation and apoptosis in kidney cortex of aging rats.Methods: Establishment of the animal aging model and groups: 50 SD female rats were applied and randomised into 5 groups: negative control group,aging model group, Heshouwuyin groups (high, middle and low dose group). There are 10 rats in each group. D-gal (dissolved by physiological saline into 6% solution) was injected into the rats abdominal cavity continually by 300mg/kg/d for 60 days. 0.9% Nacl was injected into the negative control group rats abdominal cavity in the same way. Model group and Heshouwuyin groups were poured with Heshouwuyin dose in the same day with the establishment of the aging model.The kidney cortex was taken after the rats being infused with 4% polyformaldehyde, embedded in paraffin and sectioned as usual. The thickness was 5μm. HE-staining was used to observe the conformation changes of the kidney cortex; SP immunohistochemistry was used to observe PCNA, Bax, Bcl-2, Fas, Fasl, and Caspase-3 expression of the kidney cortex; TUNEL was used to observe the apoptosis of kidney cortex cells.The rats were killed by amputating the head and the kidney cortex was taken. Western blotting was used to analyze the Bcl-2 and Bax protein content of the kidney cortex.Results:HE-staining: The conformation of kidney cortex tissue in Heshouwuyin groups improved significantly. Middle and high groups looked nearly the same as negative control group. The renal corpuscles and tubules become normal. There are more renal corpuscles.The renal tubules becomes thick and the brush border becomes obvious.SP immunohistochemistry: PCNA: The counts of PCNA positive cells of model group rats were lower than negative control group rats (P<0.05). Those of Heshouwuyin groups were higher than model group (P<0.05). The counts of PCNA positive cells of high dose group were higher than that of middle dose group (P<0.05). Bcl-2: The expression of Bcl-2 in model groups was weaker than that in negative control group (P<0.05). The Bcl-2 expression was stronger in Heshouwuyin high dose group than that of model group (P <0.05). Those of middle and low dose groups was nearly the same as model group (P>0.05). Bax: The expression of bax in model group was stronger than that in negative control group (P<0.05). The expression of Bax in Heshouwuyin groups was weaker than that in model group (P<0.05). Fas: The expression of Fas in model group was stronger than that in negative control group (P<0.05). The expression of fas in Heshouwuyin groups was weaker than that in model group (P<0.05). Fasl: The expression of Fasl in each group was different (P<0.05). The expression of Fasl in model groups was stronger than that in negative control group (P<0.05). The expression of fasl in Heshouwuyin groups was weaker than that in model group (P<0.05). Caspase-3: The expression of Caspase-3 in model group was stronger than that in negative control group (P<0.05). The expression of caspase-3 in Heshouwuyin high and middle groups was weaker than that in model group (P<0.05). Those of low dose group was nearly the same as model group (P>0.05).TUNEL: The counts of apoptotic cells of model group rats were higher than negative control group rats (P<0.05). Those of Heshouwuyin groups were lower than model group (P<0.05).Western blotting: Bcl-2: The expression of Bcl-2 protein in model group is lower than that of negative control group (P<0.05). Those of Heshouwuyin middle and high groups was higher than model group (P<0.05). Bax: The expression of Bax protein in model group is higher than that of negative control group (P<0.05). Those of Heshouwuyin groups was lower than model group (P<0.05).Conclusions:Traditional Chinese medicine Heshouwuyin can increase PCNA expression to develop proliferation of the kidney cortex. Heshouwuyin can decrease apoptosis in kidney cortex of aging rats through controling apoptosis related genes Bcl-2/Bax, Fas/Fasl and Caspase-3. Traditional Chinese medicine Heshouwuyin can protect rats kidneys and has fairly anti-aging effects.